The research also hints at why the outbreaks have confounded scientists, farmers and livestock handlers hoping to contain and prevent the disease – an effort likely complicated by the fact that the virus has an affinity for cow mammary glands rather than airways.

Tests for transmission among cows through milk machinery or through feeding of calves, and between birds and cattle through shared indoor air, didn’t show results of disease spread.

Researchers are continuing to pursue more answers, but for now the transmission mystery endures, meaning scientists can’t yet provide evidence-based recommendations for practices that would stop the spread.

“How it spreads from cow to cow becomes a very important question. We need to understand if there’s a way to change milking practices or farming practices, whatever it is, to limit cow-to-cow transmission because we think spillover is going to happen again. It’s just a matter of time,” said senior author Andrew Bowman, professor of veterinary preventive medicine at The Ohio State University.

“And right now we don’t have a great way to prevent either that spillover or cow-to-cow transmission once it happens.”

The study was published recently in Nature Communications.

Highly pathogenic H5N1 avian influenza A viruses are associated with wild birds and poultry, but since 2021 a group of these viruses of clade 2.3.4.4b have circulated globally in mammals and were first reported in U.S. dairy cattle in March 2024.

To date, 1,053 outbreaks of the originally detected virus genotype (B3.13) have been confirmed in dairy herds in 17 states, which have been brought under control through a national milk testing strategy that halted movement of herds producing milk in which the virus was detected. Current federal data suggests small numbers of infections in cows are confirmed in Idaho, Utah and Texas.

“This work has all been done in response to the unprecedented spillover of avian influenza into dairy cattle,” Bowman said. “Initially, we had no idea that cows could even be infected with influenza, let alone that the mammary gland was involved. That in and of itself was a major paradigm shift: It’s not respiratory.”

Bowman and colleagues have been following the disease since its emergence on dairy farms, reporting early this year on detection of the virus in retail milk supplies – at that time, 36.3% of samples tested were positive for H5N1 particles.

“Pasteurization is inactivating it. But once a cow’s infected, they produce high-viral-titer milk for a week-plus,” Bowman said.

Those high virus titers, or concentrations, in milk were among the results of the current work, which involved testing the effects of varied levels of viral particle inoculations into individual cow teats, where mammary glands are located.

Results showed that the smallest dose of 10 particles resulted in productive infection but fewer clinical signs compared to higher doses, as well as shedding of milk containing high concentrations of viral particles.

“They are four separate mammary glands that have a common blood supply, but in these small numbers of animals, we’re not seeing a lot of viral movement between those glands,” Bowman said. “I think that may become important to how sick a cow does or doesn’t get because if one of four glands is infected, that’s different from a severe infection in all four.”

Several experiments aimed to identify potential disease transmission routes.

Researchers transferred contaminated milking equipment directly from an infected cow to a healthy cow twice daily for 14 days. Even though infected cows had clinical signs and their milk was high in viral particles, the healthy cows exposed to contaminated equipment remained unaffected and their milk was virus-free.

Bottle-feeding calves high-viral-titer milk collected from infected cows led to minimal virus detection and inflammatory signs in the calves, suggesting transmission through feeding doesn’t spread enough material to establish an infection.

Lactating cows given an intranasal dose of the H5N1 virus did not get sick and the presence of viral RNA was minimal in nasal swabs and absent in milk, but some of their respiratory tissues showed signs of an immune response compared to uninfected cows. These cows were also co-housed with chickens that remained healthy after 17 days. The combined findings suggest airborne transmission may be minimal if it’s happening at all.

Experiments were conducted in one of only six Biosafety Level 3 facilities in the country that can accommodate large animals. There is a chance, Bowman said, that the biocontainment conditions were too sterile and airflow-controlled to resemble daily farm life, especially when it comes to airborne transmission.

And performing a limited number of tests on actual cows, rather than surrogate models, could be a “numbers game,” he said. “We see about 20% of cows on farms become clinically ill. Two transmission experiments just may not be enough.

“I think the milking equipment is something we still have to consider. The virus replicates in the mammary gland to a really high titer. It comes out in milk at a really high titer. We have common contact equipment that goes from cow to cow with high-risk material to the high-risk tissue. I think the straight line between two points would say the milking equipment still seems to be the likely route of transmission between cows,” Bowman said.

“There is still the bigger question of spillover from wild birds into cows. In waterfowl, it’s a pathogen replicating in their gut. How in the world does it go from a duck’s intestine into a cow’s mammary gland? That’s a head scratcher.”

This work was supported by the Centers of Excellence for Influenza Research and Response funded by the National Institute of Allergy and Infectious Diseases.

Cody Warren and Scott Kenney of Ohio State were co-lead authors. Additional co-authors were Carolyn Lee, Natalie Tarbuck, Hannah Cochran, Bryant Foreman, Patricia Boley, Saroj Khatiwada, Alok Dhakal, Khadijat Adefaye, Jennifer Schrock, Mohammad Jawad Jahid, Thamonpan Laocharoensuk, Raksha Suresh, Olaitan Shekoni, Erika Stevens, Sara Dolatyabi, Christina Sanders, Elizabeth Ohl, Devra Huey, Juliette Hanson, Kara Corps and Renukaradhya Gourapura, all of Ohio State, and Richard Webby of St. Jude Children’s Research Hospital.

The Plant and Animal Agrosecurity Research Facility is located at Ohio State’s College of Food, Agricultural, and Environmental Sciences Wooster Campus.  

The findings suggest influenza D has strong potential to spill over to humans, the scientists say. Some people who work with cattle, considered the primary host for this virus, have been found to have influenza D antibodies in their blood, but no active human infection has been discovered to date. 

Results showed that several genetically different strains of influenza D that were isolated from cattle and pigs, another animal host, were as effective at replicating in cells from the human respiratory tract as influenza A viruses – the types of flu linked to both modern seasonal illness and historic pandemics. 

“All of the viruses, despite their genetic distance from each other, had similar replicative capacity in these human cells and human tissues,” said lead author Cody Warren, assistant professor of veterinary biosciences at The Ohio State University. 

“It seems like there are many animal species that are susceptible to influenza D viruses, suggesting that maybe they can evolve differently in different hosts. What’s obvious is that zoonotic infections are happening. 

“This is an opportunity, if we are indeed in front of it, to invest in surveillance and basic understanding of its biology such that we could be prepared if it were to emerge in the future.” 

The study was published this week in Proceedings of the National Academy of Sciences. 

The influenza D viruses from pigs used in the study were collected as part of long-term surveillance for swine flu – a type A influenza virus – at county and state fairs led by Andrew Bowman, professor of veterinary preventive medicine at Ohio State. 

“Flu D popped up in our samples and that led us to question what the risk is at the human-animal interface,” said Bowman, a co-author of the study. 

“The currently held belief is that bovines are the natural host for flu D. So pigs may be a secondary host, or not. The fact that it was in pigs is something we were trying to understand: Is that a potential pathway for the virus to adapt into a more transmissible virus in humans? So the pig component is trying to understand what the role of the pig is with influenza D.” 

Warren’s lab began by testing influenza D’s replication in cells that mimic the human airway by differentiating patient-derived lung epithelial cells at the air-liquid interface – cellular conditions the virus would encounter naturally. 

“When the virus started growing in those just as well as influenza A, we decided to leverage another physiologically relevant system to evaluate the breadth of cell types targeted by this virus: tissues,” Warren said. 

The team compared influenza types D and A growth in both human and swine lung tissues, finding that both virus types replicated efficiently in lung tissues from both species. 

The experiments in cells and tissue also showed a key difference between the two flu types: Influenza D, despite growing well, did not stimulate a robust antiviral immune response in infected cells, while the influenza A virus did. 

Separate tests in cell cultures showed influenza D growth was restricted if the cells were first primed by the protein interferon – suggesting that human cells have potent mechanisms to restrict viral infection. 

In infected animals, influenza D generates respiratory symptoms that cattle and pigs recover from, suggesting their immune systems are at work fending off the infection. In humans, most viral infections trigger the release of interferon, which produces inflammation and raises body temperature – leading to the symptoms that make us feel sick. 

These initial findings prompt important questions: Could humans have been infected but not felt sick, meaning influenza D is not a serious health risk? Or is it so stealthy that it can hide from the immune system, leaving us unable to put up a fight? 

“Those are gaps that we don’t quite understand,” Warren said. “The virus replicates to really high levels but doesn’t elicit a robust interferon response. Would it behave differently in the body of a person versus in these cell or tissue-based systems? That’s up for debate. 

“It seems the only smoking gun we’re missing is a virus isolated from a person. There’s been evidence of a history of exposure, but not necessarily any demonstration of active infection.” 

Bowman has collected many more influenza D viruses from pigs through ongoing swine flu surveillance, and plans to analyze the samples to assess whether the viruses are changing, and how. 

“What’s giving influenza D attention now is that it’s recognized as a cause of respiratory disease in cattle, and we’ve certainly seen it cause disease outbreaks in pigs,” he said. “Causing disease in some host species makes us somewhat concerned about what it might do in humans. It may not cause disease in this particular form, but in an evolved form, there may be that potential.” 

This work was supported by the U.S. Department of Agriculture, the National Institute of Allergy and Infectious Diseases, the National Institutes of Health and the Ohio State Enterprise for Research, Innovation and Knowledge. 

Additional co-authors include Christina Sanders, Min Liu, Jovanna Fusco, Elizabeth Ohl, Natalie Tarbuck, Emily King, Devra Huey and Amanda Panfil of Ohio State; Thomas Fabrizio and Richard Webby of St. Jude Children’s Research Hospital; and Phylip Chen and Mark Peeples of Nationwide Children’s Hospital.

A new study provides the first scientific evidence that the answer is yes, showing that two species of ticks can live at least one week, and up to about three weeks, on hard-surface and carpeted floors. 

The Ohio State University study focused on lone star and Gulf Coast ticks, both of which are capable of transmitting pathogens that can make people and animals sick. Survival rates of each species were assessed in five types of common home flooring: tile, wood, vinyl, and short-pile and shag-like carpet. 

Overall, Gulf Coast ticks survived for significantly longer periods of time than lone star ticks, but in an unexpected finding, lone star ticks lived longer overall in long-pile carpet. 

“Ticks can pose a risk even in the places you least expect, such as your house,” said first author Afsoon Sabet, a PhD student in entomology at Ohio State. 

The findings underscore the necessity of taking protective measures to prevent tick bites, the researchers say. These include using tick prevention products for people and pets and, once inside, thoroughly checking an animal for ticks with a brush or a lint roller and immediately bagging clothes or putting them in the dryer. 

“The ultimate goal from a public health perspective is to reinforce that ticks brought into the home on pets or people could be a risk, and hopefully having some data around that will compel people to say, ‘OK, doing tick checks is really important,’” said co-senior author Risa Pesapane, associate professor of veterinary preventive medicine at Ohio State.   

“That way you’re not releasing ticks into your home environment where you’re relaxed and comfortable, and most definitely not thinking about tick prevention.” 

The research was published online March 13 in the Journal of Vector Ecology. 

Total reported tick-borne disease cases in the United States increased by 40% from 2019 to 2022, according to the Centers for Disease Control and Prevention. The ticks in the study, Gulf Coast (Amblyomma maculatum) and lone star (Amblyomma americanum) ticks, are commonly found in the eastern U.S. and feed on a variety of birds and mammals. Gulf Coast ticks can transmit several spotted fever diseases, and lone star ticks can transmit ehrlichiosis. In addition, a compound in lone star tick saliva can induce alpha-gal syndrome, a potentially life-threatening allergy to a molecule in red meat and other mammal products. 

For the study, Sabet tested the survival time of 90 ticks per species placed in three rounds of experiments on each of the five floor types. Ticks were individually covered with a cup so they couldn’t escape, but they could move around. Two rounds of the experiments compared survival to control ticks kept in a growth chamber to confirm experimental tick deaths were related to the flooring conditions. 

In nature, ticks feed once per life cycle. Ticks in the experiments were adults that had not yet fed. 

“Generally, what we’re talking about here is hitchhiking ticks that have come in on clothes or pets that have not yet bitten and are actively looking for a host,” said Pesapane, who also has a faculty appointment in Ohio State’s School of Environment and Natural Resources. “They would’ve been looking for a host to feed from in someone’s home.” 

Sabet monitored the ticks daily, with a few exceptions, completing over 4,600 individual observations. 

The average overall survival time of Gulf Coast ticks was about 18 days, and for lone star ticks, 11 days. The longest survival time for Gulf Coast ticks was an average of 25 days on vinyl and the shortest, 10 days on long-pile carpet. Average survival time of lone star ticks was the longest, almost 15 days, on long-pile carpet and shortest on tile – about seven days. 

The cause of death was presumed to be desiccation, or loss of moisture. Meanwhile, some of the control ticks kept at ideal temperatures and humidity levels in the lab survived for more than a year. 

“The tick life cycle is usually at least two years, typically. And the longevity of that tick is based on its ability to maintain moisture,” Pesapane said. “And certainly, the conditions in a home are desiccating conditions, but this shows that ticks are not going to die immediately. 

“Pinpointing exactly how long a tick might survive in a home is really difficult because people’s homes are going to vary in their environmental conditions. But for the first time, this study puts some guardrails on that estimate, and it shows us that ticks can survive for at least a week.” 

Samuel Ward, assistant professor of entomology at Ohio State, was an additional co-senior author of the study. This work was supported by the National Institute of Food and Agriculture, U.S. Department of Agriculture.

Researchers followed up on a 2014 study finding that Ohio’s first established population of blacklegged ticks, carriers of the bacteria that cause Lyme disease, had been detected in 2010 in Coshocton County.

At that time, the infectious bacteria were detected in 2.4% of collected blacklegged ticks and antibodies indicating prior exposure to the pathogen were detected in 20% of white-footed mice, one of the most common of several small mammal sources of wildlife infections to ticks in their nymph and larval stages. 

In the new study, the research team from The Ohio State University found that the prevalence of infection in captured ticks had increased to as much as 47.6% and in infected small mammals, up to 60.4%. 

“Our suspicion was that 10 years later, when we have robust populations of the right hosts in this ecological community in Ohio, that this pathogen vector system could really establish and take off. And that’s what this study essentially shows – that that is what happened,” said senior author Risa Pesapane, associate professor of veterinary preventive medicine at Ohio State.  

“And now Ohio has the same risk as those endemic regions in the Northeast. I think that is surprising to a lot of people because we think, sure, we have the tick and we have some of the hosts, but we’re not Connecticut. But for some parts of Ohio, we are Connecticut at this point.”

Pesapane noted that the risk for infection varies across Ohio and is highest in forested areas in the eastern and southern regions of the state. That said, the presence of blacklegged ticks has been reported in all 88 counties. 

“I think it’s really important to convey, when it comes to both personal protection and to the medical community’s awareness, that you have the same chance of getting Lyme disease in Coshocton, Ohio, and other Ohio areas with the right habitat, that you do in Lyme, Connecticut.” 

The study was published recently in the Journal of Medical Entomology.

Blacklegged ticks, also known as deer ticks, are widely distributed in the eastern United States and are a known carrier of the Borrelia burgdorferi strains of bacteria that cause most North American Lyme disease cases in people and dogs. The ticks also carry Anaplasma phagocytophilum, the pathogen that causes anaplasmosis. 

Ningzhu Bai, a PhD student in the Environment and Natural Resources Graduate Program at Ohio State, led this study for her master’s thesis. The team collected 654 ticks demonstrating questing behavior and trapped 106 small mammals. Ticks pick up the infectious bacteria from small mammals that are reservoir hosts, which infect the ticks that bite them for a blood meal. 

Deer, by the way, are not involved in the cycle of infection that occurs in ticks’ earliest life stages, but are strongly linked to blacklegged ticks because white-tailed deer are the main hosts of adult ticks that are looking for blood meals and mates. 

The researchers extracted genomic DNA from 368 ticks and from mammal tissue and blood samples to test for Borrelia burgdorferi and Anaplasma phagocytophilum. 

“It was shocking to find that infection prevalence increased from 2.4% for Borrelia burgdorferi sensu lato to almost 50% in 10 years,” Bai said. “And it was also really high for Borrelia burgdorferi sensu stricto – over 40% carried that pathogen.” 

The infection prevalence in ticks for the anaplasmosis pathogen was 15.5%. Six of the eight mammal species tested positive for infectious bacteria with prevalence of 60.4%, 19.8% and 11.9% for the same pathogens, respectively. White-footed mice and eastern chipmunks were the most frequently infected hosts. 

“When I talk to people, I like to stress this means one out of every two ticks that you might encounter in Ohio could possibly be infected,” said Pesapane, who also has a faculty appointment in Ohio State’s School of Environment and Natural Resources. 

The study provides the ecological data needed to guide where interventions could have the most impact – evaluating the effectiveness of tick-killing acaricides on small mammal reservoirs is an upcoming research plan for the Coshocton properties visited in this study. 

“Ecological interventions are challenging. That’s why we keep promoting personal protective actions, which work extremely well,” Pesapane said. Steps include wearing tick repellent and long sleeves and pants, and tucking pants into socks while outdoors; performing a thorough tick check once inside, and showering; and using preventives for companion animals. 

In Ohio, these protective tasks are worth it, she said: “If we look from 2010 all the way to 2025, there’s been a 48-fold increase in the number of Lyme disease cases in Ohio, and we are going to be considered a high-incidence state moving forward.” 

In addition to frequently collecting and studying ticks of medical concern across Ohio, Pesapane invites the public to mail ticks to her lab. She is also the faculty director of the Buckeye Tick Test, an Ohio State service that tests submitted ticks for pathogens that cause disease, including Lyme disease, for a fee. 

These submissions, and her own research, have made it clear that tick bites are not only a warm-weather threat. 

“Even on days that have snow, as long as the ambient temperature is above freezing, blacklegged ticks will be seeking hosts and biting rather indiscriminately whomever they encounter,” she said. “So year-round, there’s a tick exposure risk.” 

Andreas Eleftheriou of Ohio State was also a co-author of the study. This work was supported by the U.S. Department of Agriculture National Institute of Food and Agriculture.

While these cell surface proteins are valuable for viral entry, they also have roles in producing an antiviral response. Before that can happen, the virus manipulates a process cells use to keep themselves healthy to lower the proteins’ activity, clearing the way for unfettered viral infection. 

Though other viruses, such as HIV, are known to silence host receptors that let them into cells, Zika is unusual for having at least three of its own proteins that can get the job done, said Shan-Lu Liu, senior author the study and a virology professor in the Department of Veterinary Biosciences at The Ohio State University. 

“That’s the most interesting part: It’s amazing that not only one, but several Zika proteins can do this,” said Liu, also a professor in the Department of Microbial Infection and Immunity. “We looked at two Zika virus strains and examined three physiologically relevant cell types. With both strains, we could see the downregulation in all three cell types. It looks like this is an important mechanism.” 

The study was published May 23 in Proceedings of the National Academy of Sciences. 

The Zika virus, transmitted to humans primarily by Aedes aegypti mosquitoes, has caused infectious outbreaks in Africa, the Americas, Asia and the Pacific since 2007, according to the World Health Organization. Though cases have declined globally since 2017, virus transmission continues at low levels in the Americas and other endemic regions. 

A large epidemic in Brazil in 2015 led to confirmation of a link between Zika infection during pregnancy and babies born with congenital problems including microcephaly, or smaller than normal head size. While most infected people develop no or only mild symptoms, the virus is also associated with Guillain-Barré syndrome, neuropathy and myelitis (spinal cord inflammation) in adults and older children. 

Previous research has shown that specific cell surface proteins known as PS receptors are important entry points for many viruses, including Zika. This study focused on two of these proteins, known as AXL and TIM-1, that had previously been linked to Zika infection. In this work, Liu and colleagues set out to explain how Zika sustains infection after gaining entry through AXL and TIM-1. 

The team completed cell culture experiments using African and Asian strains of Zika virus in three types of cells related to respiratory, reproductive and neurological systems targeted by the pathogen: human cells that line the lungs, embryo-supporting cells called trophoblasts and glioblastoma brain cancer cells. 

Experiments showed that both AXL and TIM-1 were downregulated on the three types of cells after infection by Zika. The researchers expected to find this suppression occurred through two common protein degradation processes, but found instead that the Zika virus makes use of a cellular self-preservation routine: autophagy. 

“Autophagy is a fundamental physiological mechanism to conserve cellular processes by degrading host components. It’s also called self-eating – the host needs to remove their own damaged organelles or misfolded proteins because they’re not good for the host,” said Liu, also associate director of Ohio State’s Center for Retrovirus Research and a program co-director of the Viruses and Emerging Pathogens Program in Ohio State’s Infectious Diseases Institute. 

In this case, the virus’s infectious process manipulated the host cells into suppressing their own protective proteins – a viral adaptive tactic that allows Zika to control its own destiny. 

Without this suppression, AXL and TIM-1 would begin producing inflammatory molecules as part of an antiviral response. Their normal level of facilitating viral entry could also enable more Zika particles to access already-infected cells, setting up a competitive scenario called a superinfection – something viruses want to avoid because the overcrowding threatens to kill cells, which kills infecting pathogens. 

Further experiments identified three Zika proteins that prompt host cell autophagy, all of which are located on the virus’s membrane. 

“Normally those proteins mediate viral entry or are involved in viral replication, but they’re also responsible for this downregulation – kind of a new function, which is not so surprising because viruses encode something that’s important for them, either for their own replication or to modulate the host,” Liu said. 

Though further research is needed to know for sure, there is a chance this mechanism is relevant to the Ebola virus, which uses the TIM-1 protein to access host cells, or to other pathogens in the same flavivirus family as Zika, including West Nile, yellow fever and dengue viruses. 

“The bottom line is this speaks to the co-evolution of viral-host interactions. The more important a host factor is to a virus, the more a virus is going to do to take control of it,” Liu said. “Understanding these mechanisms is an important part of being prepared for emerging or reemerging viruses that cause infectious diseases.”

This work was primarily conducted by Jingyou Yu, a former graduate student in the Liu lab and now a principal investigator at the Guangzhou National Laboratory in China. Additional contributions were made by Yi-Min Zheng, a senior scientist, and Pei Li, a postdoctoral fellow in the Liu lab. This work was primarily supported by an Ohio State fund and the National Institutes of Health. 

Additional co-authors are Megan Sheridan, Toshihiko Ezashi and R. Michael Roberts of the University of Missouri.

The protective effect was enhanced by delivering the antibody in a nasal spray that disperses these molecules throughout the respiratory tract, where they stick to the slippery mucus lining to lie in wait for invading viral particles. 

The engineered molecule is based on the IgM version of an immunoglobulin antibody, the generic first line of antibody defense against infection – but with an added structural feature that attaches to a very specific location on the influenza A surface to neutralize the virus. 

“It’s a better platform with a better antibody. If we can prepare the respiratory environment with this enhanced engineered molecule, it can capture and intercept the virus in an early stage,” said co-lead author Kai Xu, assistant professor of veterinary biosciences at The Ohio State University. “This would be a therapy that not only can be used for prevention of seasonal flu – it potentially could also be used against pandemic strains in the future.” 

The study was published April 29 in Nature Communications. 

Influenza A viruses are divided into subtypes based on characteristics of two major surface proteins, hemagglutinin (HA) and neuraminidase (NA) – as in, for example, the strain annually circulating in people, H1N1, and a less common H3N2 virus that co-circulates during flu season. 

Previous work has shown that monoclonal IgG antibodies engineered to target the influenza A HA protein lose potency as the virus mutates its way out of their crosshairs. 

These antibodies, mimicking mature natural flu-fighters of the adaptive immune system, attach to a precise section of the surface protein to disable the virus’s ability to bind to a host cell receptor and enter the cell to make copies of itself. But once viruses figure out where they’re being targeted by antibodies, they mutate those sites to escape recognition by the immune system. 

Xu and colleagues theorized that building an anti-flu antibody based on the IgM isotype (an early, first-line immune responder known for its pentameric shape), featuring more structural sites capable of binding to the HA protein as well as an IgG-level tight and precise target, would increase the antibody’s neutralization capacity.  

The team constructed and evaluated 18 IgM antibodies based on existing IgG antibodies known to target different segments of the HA protein, landing on one IgM version that was superior in cell culture studies at neutralizing a panel of H1N1 and H3N2 flu viruses that have been circulating in people for at least 50 years. 

“The advantage is that we utilized the multivalency feature of IgM combined with the potency of a mature broadly neutralizing antibody,” Xu said. “Multivalent means that this molecule has multiple arms to capture the virus, so the virus has very little chance to escape. 

“And because IgM is a large molecule, when it binds to the viral surface it effectively covers the surface, so it prevents receptor engagement.” 

Experiments showed that even if a mutation affected this IgM antibody’s strong attraction to a specific HA binding site, the antibody’s large size and multivalency could help it outpace viral escape attempts. 

“By enhancing the capacity for it to recognize the mutated or differentiated viral surface regions, it can cross-protect against different influenza virus infections and can also work even after mutations occur,” Xu said. 

In mouse experiments, researchers delivered a single dose of the IgM antibody in a nasal spray, intending to place a layer of the engineered antibody across the mucosal surfaces of the nasal cavities and lungs. 

“The primary purpose is to refuse viruses before they can get into a host cell,” Xu said. “The mucosal immunity is more efficient than systemic immunity at intercepting, so the initial infection can be reduced or eliminated.” 

Researchers followed the preventive nasal spray with lethal doses of H1N1 and H3N2 flu viruses. Compared to control animals that did not receive preventive treatment, mice receiving the IgM antibody had better outcomes: No mice got sick from the H3N2 strain, and most survived the H1N1 infection and recovered from symptoms. 

The molecules remained on the mouse mucosal surface for seven days, an indication that a single spray could last for weeks in humans. 

“The adhesiveness of the molecule extends its protection window, and that’s one advantage of this strategy,” Xu said. 

Though this work focused on historic and seasonal versions of influenza A, Xu said the team of structural biologists, immunologists and virologists see the findings as a proof of principle to help establish the IgM platform as a potential therapeutic technology for different viruses – including bird flu, an influenza A H5 virus – and even non-viral diseases like cancer. 

Xu co-led the study with Xinli Liu of the University of Houston, Tong-Ming Fu of IGM Biosciences Inc. and Zhiqiang An of the University of Texas Health Science Center at Houston. Additional co-authors from those institutions and the National Institute of Allergy and Infectious Diseases also worked on the study. 

This research was supported by grants from the National Institutes of Health, IGM Biosciences and the Welch Foundation.

The AAAS Fellowship, recognizing scientifically or socially distinguished efforts to advance science or its applications, is one of the most prestigious honors a U.S. scientist can receive. Fellows are elected by their academic peers.  

“Ohio State’s newly elected Fellows have devoted their professional lives to protecting health, investigating complex principles of physics, and analyzing social behavior and labor economics,” said Peter Mohler, executive vice president for research, innovation and knowledge. “The American Association for the Advancement of Science Fellowship is an important acknowledgment of the key role scientists play in solving critical problems and improving lives.”   

The 2024 class consists of 471 scientists, engineers and innovators spanning 24 scientific disciplines.   

Ohio State’s newest Fellows represent the colleges of Arts and Sciences; Food, Agricultural, and Environmental Sciences; Medicine; and Veterinary Medicine. This year’s class, joining over 540 Ohio State scholars who have been granted this lifetime recognition since 1874, are:   

John Beacom, professor of physics and astronomy. For distinguished contributions to the fields of nuclear and particle astrophysics, particularly regarding neutrinos. 

Pierluigi (Enrico) Bonello, professor of molecular and chemical ecology of trees in the Department of Plant Pathology. For distinguished contributions to the fields of forest pathology, chemical ecology, tree disease resistance and early detection of tree infections. 

Prosper Boyaka, professor and chair of veterinary biosciences. For distinguished contributions to the field of mucosal immunology, particularly in the field of vaccinology to develop alternative novel strategies to optimize protective immunity to pathogens at mucosal surfaces. 

David Robert Cole, professor of earth sciences and Ohio Research Scholar. For distinguished contributions to the field of subsurface mineralogy and geochemistry from the molecular to the geological formation scale. 

Ian Hamilton, professor of evolution, ecology and organismal biology and mathematics. For distinguished contributions to the fields of behavioral ecology and complexity science in both theory and empirical studies. 

Chun Ning Jeanie Lau, professor of physics. For distinguished experimental contributions to the field of 2D van der Waals materials, leading to new insights into the thermal, mechanical and electronic properties, and emergent phases in few-layer graphene. 

Roland Kenji Kawakami, professor of physics. For seminal contributions to spintronics and magnetism in two-dimensional materials, and for the investigation of epitaxial magnetic films and heterostructures using molecular beam epitaxy and surface-sensitive characterizations. 

Abhay Satoskar, professor of pathology and microbiology. For distinguished contributions to the field of parasitology, particularly mechanisms of immunity and immune evasion in leishmaniasis and development of novel therapies and vaccines for this disease. 

Bruce Weinberg, professor of economics. For distinguished contributions to the field of labor economics, particularly for a multidisciplinary approach to studying creativity and innovation, socioeconomic determinants of youth outcomes and technological impact on wage structure. 

Fellows are recognized for their achievements across disciplines, from research, teaching and technology to administration in academia, industry and government, to excellence in communicating and interpreting science to the public. 

The 2024 class of Fellows will be honored at a forum in Washington, D.C., in June.

In a new study, researchers identified a protein in mammals that welcomes arteriviruses into host cells to start an infection. The team also found that an existing monoclonal antibody that binds to this protein protects cells from viral infection. 

Arteriviruses circulate broadly in many types of mammals around the world that serve as natural hosts – such as nonhuman primates, pigs and horses – but so far have not been detected in humans. 

The researchers’ aim is to better understand mechanisms of arterivirus infection to get a handle on how high the infection risk is for humans and what preparation may be needed should a spillover occur in the future. 

“It’s important to consider that since we have no known arteriviruses infecting people that we’re essentially immunologically naïve, so we can’t rely on preexisting immunity to help us,” said co-lead author Cody Warren, assistant professor of veterinary biosciences at The Ohio State University. 

Warren co-led the work with Adam Bailey, assistant professor of pathology and laboratory medicine at the University of Wisconsin-Madison. The study was published recently in Nature Communications. 

Many natural hosts of arteriviruses have no signs of disease, but the virus that infects swine can cause pneumonia, as well as abortions in pregnant pigs, and other strains can cause hemorrhagic fever or encephalitis when they switch animal hosts. 

These viruses also have the unusual ability to maintain long-term infections and become more virulent when they find new hosts – which gives them time to evolve and improve their chances of transmission. 

The research team set out to find proteins in mammals that arteriviruses use as receptors to gain entry to host cells and make copies of themselves. Bailey used genome-wide CRISPR-knockout screening technology to identify specific genes that, when disrupted, rendered cells resistant to viral infection. Such genes would then be considered essential to the viral infection process. The unbiased screen identified two genes, FCGRT and B2M, whose protein products come together to form the FcRn receptor (neonatal Fc receptor) that is expressed on the surface of cells. 

The FcRn receptor molecule has a specific role in shuttling antibodies across the placenta to a fetus, but is also present in immune cells and cells that line blood vessel walls – both of which are targeted by arteriviruses. 

Results from this study demonstrated that FcRn is used for host cell entry by at least five arteriviruses that infect monkeys, pigs and horses, respectively: three diverse strains of simian arteriviruses, porcine reproductive and respiratory syndrome virus 2 (PRRSV-2), and equine arteritis virus (EAV). 

Knocking out the major component of the FcRn complex – the FCGRT gene – from host cells blocked viral infection, and pre-treating cells with a monoclonal antibody against FcRn protected against infection. 

There was also a genetic twist to this story: Some mammal hosts were less susceptible to arterivirus infection based on differences in their species-specific FcRn’s sequence, meaning that in some cases, this protein will function as a barrier to cross-species infections. 

“Chimpanzees and humans have pretty much all the same genes, but the sequence of those genes is slightly different,” Bailey said. “All mammals have the FcRn receptor, but their ability to support infection with a given arterivirus may vary.” 

The CRISPR screen also identified a gene encoding another surface protein, CD163, which Warren and colleagues previously found to be a gatekeeper for an arterivirus called simian hemorrhagic fever virus (SHFV) to infect a cell. 

A series of experiments in different cell types and using multiple viral strains in the new study showed that CD163 does have a role in infection by most arteriviruses, but it cannot act alone – interaction with FcRn is also key to facilitating arteriviral infection of host cells. 

Spelling out these arterivirus infection steps is an important milestone, the researchers said. 

“If we’re looking at virus biology, one of the most important things we can understand is entry mechanisms. Because if you can stop the ability of a virus to infect a cell through disrupting that initial virus-receptor contact, now you have a potential therapeutic strategy,” Warren said. 

One of those “disruptors” could be blocking the receptor – so showing that an existing monoclonal antibody can stop viral infection in cells is also a plus for scientists examining viruses through a lens of pre-pandemic preparedness. 

“If one of these viruses emerged in humans, I believe we’d be in big trouble,” Bailey said. “So that is the motivator for me.” 

This work was supported by National Institutes of Health grants, University of Wisconsin-Madison startup funds, the G. Harold and Leila Y. Mathers Foundation, and the Burroughs Wellcome Fund Pathogenesis of Infectious Disease Program. 

Co-authors included Teressa Shaw, Kylie Nennig, Xueer Qiu, Devon Klipsic and Igor Slukvin of UW-Madison; Devra Huey, Makky Mousa-Makky, Jared Compaleo, Fei Jiang and Haichang Li of Ohio State; Aadit Shah of Stanford University; Raymond Rowland of the University of Illinois Urbana-Champaign; and Meagan Sullender and Megan Baldridge of Washington University in St. Louis.

The Rocahepevirus ratti strain is called “rat HEV” because rats are the primary reservoir of the virus. Since the first human case was reported in a person with a suppressed immune system in Hong Kong in 2018, at least 20 total human cases have been reported – including in people with normal immune function. 

People infected with rat HEV did not report exposure to rats, leaving the cause of infection undefined. The suspected cause during other human HEV infections, in many cases, is consumption of raw pork – making it a potential route for rat HEV as well. 

Researchers at The Ohio State University found that a strain of rat HEV isolated from humans could infect pigs and was transmitted among co-housed animals in farm-like conditions. Rats are common pests in swine barns – suggesting that the pork production industry may be a setting in which rat HEV could make its way to humans. 

“We always want to know which viruses might be up and coming, so we need to know the genetics behind this virus in the unlikely event something happens in the United States that would enable rat HEV to expand,” said senior author Scott Kenney, an associate professor of animal sciences and veterinary preventive medicine at Ohio State based in the Center for Food Animal Health at the College of Food, Agricultural, and Environmental Sciences’ Wooster campus.  

The study was published recently in PNAS Nexus. 

Hepatitis E is the leading cause of the acute viral liver infection in humans worldwide, mostly in developing regions where sanitation is poor. The virus is also endemic in pigs in the United States – though it is present mostly in liver rather than muscle, and is killed when the meat is cooked.  

Past studies testing the cross-species infectiousness of rat HEV showed the strain used in experiments did not infect non-human primates. 

“It dropped off the radar for six or seven years because it was thought not to be a human pathogen. And now it’s infecting humans, so we need to figure out why,” Kenney said. 

One strain linked to human disease is known as LCK-3110. First author Kush Yadav, who completed this work as a PhD student in the Center for Food Animal Health, used the viral genomic sequence to construct an infectious clone of LCK-3110. 

The team first showed the cloned virus could replicate in multiple types of human and mammal cell cultures and in pigs. Researchers then injected pigs with an infectious solution containing the LCK-3110 strain or another HEV strain present in pigs in the U.S., as well as saline as a control condition. 

Viral particles in the blood and feces were detected one week later in both groups receiving HEV strains, but levels were higher in pigs infected with rat HEV. Two weeks later, co-housed pigs that received no inoculations also began to shed rat HEV virus in their feces – an indication the virus had spread through the fecal-oral route. 

Though infected pigs’ organs and bodily fluids were also positive for viral RNA, the animals did not show signs of feeling sick. Previous research suggests rats don’t have clinical symptoms, either. 

Even so, the rat HEV virus was detected in cerebrospinal fluid of infected pigs – a finding that aligns with growing concern that various strains of HEV that infect humans can harm the brain. One human death linked to rat HEV was caused by meningoencephalitis. 

“HEV is gaining importance for neurological disorders, and a lot of the research now points toward how neuropathology is caused by the hepatitis E virus,” Yadav said. “And even though we have a small number of known human cases, a high percentage of them are immunosuppressed. That means transplant recipients in the United States could be at risk of infection by general HEV as well as rat HEV. 

“Research could now focus on whether pork liver products contain rat HEV and explore food safety procedures to block the disease.”

Yadav is now a postdoctoral researcher in the Virginia-Maryland College of Veterinary Medicine at Virginia Tech. Co-authors of the study, all from Ohio State, were Patricia Boley, Carolyn Lee, Saroj Khatiwada, Kwonil Jung, Thamonpan Laocharoensuk, Jake Hofstetter, Ronna Wood and Juliette Hanson.

Since that study’s publication, scientists at The Ohio State University have been exploring how the SARS-CoV-2 virus that causes COVID-19 prompts this human molecule’s destructive activity, and outlined the series of steps needed to fully describe what’s going on – as well as potential strategies to stop the damage. 

The grant from the National Institutes of Health (NIH) will fund their five-year pursuit of definitive answers and development of new ways to treat acute SARS-CoV-2 infections and, ideally, fend off long COVID. The award is the largest of its kind funding infectious diseases research at Ohio State. 

The 2022 published research showed in mice infected with SARS-CoV-2 that blocking this molecule, an enzyme called caspase 11, resulted in lower inflammation and tissue injury and fewer blood clots in the animals’ lungs. The researchers also found that the human version of the enzyme, called caspase 4, was highly expressed in COVID-19 patients hospitalized in the ICU – confirming the molecule’s link to severe disease. 

The new work funded by the NIH will extend the investigation beyond the lungs based on predictions that in response to the viral infection, caspase 11 has compounding effects in multiple cells: driving up inflammation in the body and brain, interfering with the immune response and leading to clots in small blood vessels. The team will also explore how SARS-CoV-2 infection shapes host and viral RNA modifications, which occur during gene activation and alter cell functions. 

Many of the affected cells being investigated are related to the immune response – both the innate response, the body’s first line of defense against any foreign invader, and the adaptive response, which is a later, specific response to a given pathogen. Researchers will also examine cells that line organ surfaces and blood vessel walls (epithelial and endothelial cells, respectively) as well as RNA modifications.   

“When you pull it all together, offering the scientific community a basic understanding of what happens to every cell and every organ during SARS-CoV-2 is an achievement in itself,” said Amal Amer, professor of microbial infection and immunity in Ohio State’s College of Medicine and the contact principal investigator on the grant. 

“Once you know the mechanism, then you can design what to target, where to target it and how to target it in order to reduce the damage being done,” Amer said. “And this is especially needed for long COVID – it may be in the brain, it may be in the muscles, it may be in anything and everything – and that’s an important aspect of the disease.” 

The federal award is a multi-principal investigator (PI) research program project grant composed of three scientific projects and four core activities (see descriptions below). Along with Amer, Estelle Cormet-Boyaka and Jianrong Li, both professors of veterinary biosciences at Ohio State, are MPIs on the initiative. The group also involves other experts from Ohio State, Nationwide Children’s Hospital and the University of Chicago. 

Amer is an expert in innate immunity who has been studying the class of molecules called inflammasomes for years. She will lead studies of the role of caspase 11, which is an inflammasome-related enzyme, in causing inflammation in the brain and lung that drives the damaging interplay between the innate immune response and blood clot formation. 

Cormet-Boyaka is an expert in lung biology, physiology and pathology, and will oversee studies of the multiple cell types whose functions are influenced, mostly negatively, by the presence of caspase 11 during SARS-CoV-2 infection. 

“In addition to studying mice, we’ll also be using human cell samples that enable us to dissect mechanisms at the cellular level,” she said. “Having access to human primary epithelial cells is a strength because those are the cells that the virus infects first.”

Li is a virologist who has been studying respiratory viruses for more than 25 years. He and colleagues will map SARS-CoV-2-induced RNA modifications in host cells and work on experimental inhibitors of molecules that trigger the RNA changes as a strategy to suppress the virus’s ability to make copies of itself in infected cells. The team will develop and test RNA modification and caspase 11 blockers to synergistically reduce SARS-CoV-2 replication, pathology and clotting, protect tissue and prevent the over-production of pro-inflammatory proteins called cytokines. 

“The two major causes of death from COVID are the cytokine storm and uncontrolled virus replication,” Li said. “If we inhibit only one of these, it’s not ideal. If we inhibit both, that can lead to a better therapeutic approach.” 

Based on data collected since the 2022 study, blocking caspase 11 remains a chief goal – but getting the right drug formulated to do it requires the information that will be uncovered by the combined projects. Though mice lacking the gene to make caspase 11 look and act normal, the research team wants to zero in on inhibitors that pose the lowest risk for side effects. 

“When you inhibit caspase 11, you get rid of many cytokines, which damage the lung tissue and the blood-brain barrier and brain tissue,” Amer said. “Combining that together with stopping viral replication is going to be very effective at reducing deaths and severe illness from SARS-CoV-2 infection, and reducing the post-infection symptoms experienced by people with long COVID.” 

Conducting simultaneous studies on different tracks will accelerate the pace of the research, said Prosper Boyaka, chair of veterinary biosciences at Ohio State and the leader of one of the three projects. An expert in the adaptive immunity that is a major player in anti-viral immunity, Boyaka will also provide a strategy to tackle immune cells called neutrophils to avoid exacerbated immune responses. 

“Long COVID is extremely complex. And the way we do science is to understand mechanisms – but because of our collective own expertise and the tools we have, we will approach one area or one question at a time,” he said. “Having a team like this one allows us to look at those interactions and processes at the same time by experts in different fields, which makes it more likely we will capture information that would be difficult to capture otherwise. That’s why I think the outcome is likely to be more beneficial than if each project were done individually or in isolation.” 

Xiaoli Zhang, an associate professor-clinical in the Department of Biomedical Informatics and Center for Biostatistics at Ohio State, is a team scientist in a broad range of biomedical research areas, mainly in cancer and microbial infection and immunity. With expertise ranging from experimental design to biostatistics and bioinformatics data analysis and modeling, she will oversee all bioinformatic and statistical analysis in the project grant.

Amer noted that program grants are very competitive, and successful applications are those that prove the PIs have a track record of working together on significant research – an indication that the team will work together efficiently for the duration of the grant.

“Being at Ohio State, we have people specializing in everything we needed for this grant, and we provided a huge list of publications going back 10 years showing we have continuously worked together and published together on cutting-edge science,” she said. “And the NIH was convinced that this group is the one that can do this.”

Grant title: “Role of the non-canonical inflammasome in SARS-CoV-2-mediated pathology and coagulopathy.”

• Project 1: Role of caspase 11 in SARS-CoV-2-induced lung pathologies and long-term immune protection (Project Leader: Prosper Boyaka; Co-Investigators: Estelle Cormet-Boyaka, Jacob Yount)
• Project 2: Caspase 11-dependent immunothrombosis and neuroinflammation during SARS-CoV-2 infection (Project Leader: Amal Amer; Co-Investigators: Stephanie Seveau, Andrea Tedeschi)
• Project 3: Caspase 11-dependent RNA modifications and their Role in Multi-Organ Pathologies (Project Leader: Jianrong Li; Co-Investigators: Mark Peeples, Chuan He)
• Administrative Core (Core Leader: Amal Amer; Co-Investigators: Estelle Cormet-Boyaka, Jianrong Li)
• Biostatistics and Bioinformatics Core (Core Leader: Xiaoli Zhang; Co-Investigators: Maciej Pietrzak, Amy Webb)
• Biological Reagents and Infection Core (Core Leader: Jianrong Li; Co-Investigator: Mark Peeples)
• Cell Derivation and Maintenance Core (Core Leader: Estelle Cormet-Boyaka; Co-Investigator: Santiago Partida-Sanchez)

Veterinary researchers analyzed fecal samples from fat cats as the animals lost and maintained weight over the course of four dietary changes, including strict calorie reduction. The team found that food-related changes to the cats’ gut microbiome – the assortment of bacteria and molecules those bacteria produce and consume – have striking similarities to dietary effects on the gut previously seen in humans. 

Though there is still a lot to learn, the findings place pet cats at the top of the list of animals whose gut bacteria may tell us a lot about our own – and whether gut microbe-based therapy could be one way to battle obesity. 

“Animals share our beds. They share our ice cream. There are all these things that people do with their pets that highlight they are a naturally occurring disease model with similar environmental exposures as humans,” said lead study author Jenessa Winston, assistant professor of veterinary clinical sciences at The Ohio State University.

“Being able to see changes in cats that come up in the context of obesity and type 2 diabetes in people makes them a really good model to start looking at more microbiome-directed therapeutics for obesity in humans if we’re seeing a similar shift,” she said. “Microbes we saw in this study also come up again and again in human studies – and clearly, people aren’t eating cat chow, right?” 

The study was published recently in the journal Scientific Reports. 

An estimated 60% of cats are obese or overweight in developed countries, and more than 2 in 5 adults in the United States have obesity, according to the Centers for Disease Control and Prevention. 

At Ohio State, Winston oversees two large clinical trials exploring the potential for fecal transplants from lean dogs and cats to help their overweight pet peers lose weight.   

“My lab is focused on how we can harness the therapeutic power of microbes,” she said. “In order to do that, we have to understand how disease states may be different from health so that we can better try and figure out and target, mechanistically, changes that occur in the microbiome.” 

In this study, researchers fed seven obese cats a four-phase diet over 16 weeks: free-feeding of commercial cat food for two weeks, free-feeding of a specially formulated weight-loss diet for one week, calorie-restricted feeding of the weight-loss diet to achieve 1-2% body weight reduction per week for 11 weeks, and a return to the original maintenance diet. 

The analysis of fecal samples taken during the different diet phases focused on changes in the presence of short-chain fatty acids metabolites, molecules produced by bacteria during digestion. Fatty acids are of interest because they prompt specific types of communication between gut microbes and tissue in the rest of the body – including hormonal signals that can relate to inflammation and insulin resistance. 

The team found that the abundance of a short-chain fatty acid called propionic acid – shown in other mammals to regulate appetite, reduce fat accumulation and protect against obesity and diabetes – was increased in feces when the cats were losing weight on the calorie-restricted diet. Greater propionic acid composition was associated with an increase in the bacterium Prevotella 9 copri. Though direct production of propionic acid by Prevotella couldn’t be determined in this study, the finding that these increases occurred at the same time, when cats were dropping weight, was intriguing. Previous research has linked Prevotella 9 copri in the human gut to weight loss and better blood sugar control. 

“When the cats are on the special diet formulated for weight loss, propionic acid goes up and stays high, and then goes back down when they’re put back on the maintenance diet. So it really is a dietary change,” Winston said. “This paper highlights that when we calorie-restrict cats that are obese, we can alter their microbial ecosystem – and those community shifts that we see likely correlate with some metabolic outcomes. 

“I think that those parallels that we’re seeing in how the ecosystems change in a similar way is helpful,” she said. 

The gut microbiome’s precise role in mammal obesity remains a mystery, but Winston said decades of evidence suggests these organisms and molecules they produce are part of the problem behind what is now known to be a very complex disease. This study’s findings in felines suggest the pet cat gut profile could provide meaningful answers for both species, she said. 

The weight-loss diet in the study was provided by Nestle Purina, which also provided funding for the project. Winston and some co-authors are paid speakers for Nestle Purina. 

Co-authors include John Rowe, Valerie Parker and Adam Rudinsky of Ohio State; Katie McCool of North Carolina State University; Jan Suchodolski, Rosana Lopes and Jörg Steiner of Texas A&M University; and Chen Gilor of the University of Florida.

Adam Berinsky, the Mitsui Professor of Political Science at MIT, opened the meeting Monday night with a lecture about misinformation – and ways to combat it. He has studied the problem in the context of politics, but the work is easily adapted to virology in an age of heightened anti-science sentiment and lack of trust in experts of all kinds. 

ASV President Anne Moscona, who selected the keynote speaker, urged audience members to consider working alongside Berinsky to address misinformation that casts doubt on scientific evidence, which she called “a problem so worrisome to our community.” 

Ohio State expertise 

Shan-Lu Liu, a virology professor at Ohio State and co-director of the Viruses and Emerging Pathogens Program of the Infectious Diseases Institute, chaired the meeting’s organizing committee. Liu, a longtime HIV researcher who more recently has led numerous studies of SARS-CoV-2 variants’ immune evasion, transmissibility and infectivity, has been named president-elect of ASV at this year’s meeting and will become president of the society in 2025. 

The conference blends symposium presentations by leaders in a range of virology disciplines with poster sessions and dozens of trainee workshops on science, career development and communication. 

Linda Saif, Distinguished University Professor in Ohio State’s Center for Food Animal Health with faculty appointments in animal sciences and veterinary preventive medicine, opened Wednesday morning’s symposium with detailed documentation of coronaviruses’ ability to spill over from animals to humans – which has been happening for centuries. An international expert on coronaviruses, Saif described the wily strains as diverse infectious agents that are skilled at accommodating change thanks their large RNA genome. 

Like the coronavirus that leapt from wild animals to cattle and from cattle to poultry that Saif’s lab was the first to document in 1995, the virus that causes COVID-19 is known to infect mink, hamsters, cats and white-tailed deer. She’s among a team of researchers who found that white-tailed deer were infected by humans with SARS-CoV-2 across Ohio and that the animals function as reservoirs promoting ongoing viral mutation. 

“If we end up with a wildlife reservoir, will it persist, adapt and mutate in wildlife with new variants going back to humans, livestock or other animals?” Saif said. “The concern is, will we end up with a scenario with SARS-CoV-2 where we have two-way transmission to wildlife species and back into humans?” 

The World Health Organization has predicted the next pandemic will be caused by what it calls “Disease X” – and some coronaviruses currently circulating in animals are potential candidates, she said. 

The studies detecting SARS-CoV-2 in Ohio deer were co-led by Andrew Bowman, professor of veterinary preventive medicine at Ohio State, who presented an update on that research program at the meeting. Bowman has also led influenza surveillance at the swine-human interface in commercial settings and agricultural fairs for years, and recently made news by finding H5N1 (bird flu) viral RNA in over a third of commercial milk products he and a graduate student bought in the Midwest based on a hunch that viruses detected in dairy cows were making their way to grocery stores. 

The annual conference presents “an important opportunity to educate the public about virology research,” said Liu, also associate director of the Center for Retrovirus Research and professor of veterinary biosciences. “And we expect that attendees will also benefit from Adam Berinsky’s insights about the general phenomenon of misinformation.” 

Fighting misinformation 

Misinformation that circulated during the COVID-19 pandemic lockdown – conspiracy theories that the illness’ severity was exaggerated or that the virus was deliberately released for sinister reasons – persists to this day. Berinsky, who has extensively studied Americans’ beliefs in false political rumors (such as claims Barack Obama wasn’t born in the United States), has found that rumors are “sticky” – countering them is not just about sharing the facts, but requires a messenger that people will be willing to trust. 

His research has found that about 70% of people will endorse at least one false rumor when given a list to choose from and only 30% of people reject them all, and that misinformation correction should focus primarily on reaching the undecideds – people who aren’t sure about what’s true. His work has also shown that correcting misinformation is not a “one-and-done” endeavor – it requires repeated messaging as well as a series of strategies targeting people who share fake information online. 

Working with Facebook and Google, Berinsky and colleagues advanced interventions encouraging people through “accuracy nudges” to think before they share information – advising users to evaluate information before sharing it and investigate its sources – or even asking them how important it is to them to share only accurate information. 

“Most people don’t want to share bad information,” he said. “We want to remind them of this.” 

They found in experiments that at the individual level, these nudges turned the knob toward more sharing of true information. Though the prompts won’t stop people who are posting lies maliciously, the added “friction” makes it a little more difficult for casual users to do an easy thing that has negative effects. 

A megastudy of nine interventions designed to reduce online sharing of misinformation revealed that all of them work – but each one’s effect is relatively small, on the order of 2 to 3 percentage points. Hence, the frame of mind in combating misinformation should not be a search for a single solution, but deployment of the best bundle of solutions depending on the context. 

“There’s been this long search for a magic bullet. It doesn’t exist,” Berinsky said. “But don’t give up on the enterprise.”

Hepatitis E (HEV) is the leading cause of the acute viral liver infection in humans worldwide, mostly in developing regions where sanitation is poor. The virus is also endemic in pigs in the United States – though it is present mostly in organs rather than muscle, and is killed when the meat is cooked. 

Because HEV has been linked to fatal pregnancy complications and reports of male infertility in the developing world, researchers at The Ohio State University explored its infectivity in pigs, whose reproductive anatomy closely resembles that of humans. 

After inoculating pigs with HEV, the team found the virus circulated in blood and was shed in feces, meaning that the pigs were infected, but they had no clinical symptoms –asymptomatic cases are common in humans, as well. Results also showed HEV was present on the head of sperm cells, and that these same viral particles could infect human liver cells in culture and begin replicating. 

“Our study is the first one demonstrating this association of hepatitis E virus with the sperm cell,” said first author Kush Yadav, who completed the work as a PhD student in Ohio State’s Center for Food Animal Health. “Our future studies will be directed toward understanding the association between hepatitis E virus and the sperm head more mechanistically, and using animal models to see whether sexual transmission of the virus is happening – because in the human setting, we still don’t know.” 

The study was published recently in the journal PLOS Pathogens. 

Sexually transmitted organisms are those able to find safe haven in testicles, where they are protected by a blood-testis barrier that immune cells cannot cross. In addition to pregnancy and reproductive disorders linked to HEV, there are signs it can also lead to pancreatic and neurological disorders in humans. Historically, clinical infections – even in pregnant people – have been assumed to be traced to fecal-oral transmission. 

Yadav works in the lab of senior study author Scott Kenney, an associate professor of veterinary preventive medicine at Ohio State based in the Center for Food Animal Health at the College of Food, Agricultural, and Environmental Sciences’ Wooster campus. 

Kenney studies HEV and other viruses in animals – especially those capable of infecting humans. This work is an offshoot of a larger study in pigs examining HEV strains that are resistant to antivirals. 

Using fluorescence microscopy in this new study to examine pig semen 84 days after inoculation with HEV, Yadav detected viral particles associated with at least 19% of sperm cells collected from the infected swine. 

“We can’t say they’re sitting on the outside or inside the sperm cells,” he said. “We don’t know if the hepatitis E virus can actually complete a replication cycle in the sperm head, so we’re thinking sperm is more of a carrier than a susceptible cell.” 

The study also showed that HEV’s presence correlated with damaged sperm – potentially altering their structure and decreasing their ability to move through seminal fluid. That said, the researchers can’t yet say that these changes directly translate into fertility problems, though the link between HEV infection and human infertility suggests this could be the case. 

“In between 20 and 50% of documented male infertility cases, they don’t really know what’s causing it,” Kenney said. “So we’re pushing for more screening for hepatitis E virus as a potential cause for those cases.” 

The current findings also form the basis for screening the sexual partners of pregnant women who test positive for HEV, Yadav said, even though scientists haven’t yet established sexual transmission is possible. 

There are implications for the swine industry, as well. Most commercial pig litters are produced by artificial insemination, with donor sperm distributed broadly from large breeder facilities. 

“That could be part of the problem with HEV being endemic across the country, and raises a question about whether it causes a reduction in the reproductivity of the pigs,” Kenney said. “Because HEV doesn’t cause enough damage to pigs to limit production profitability, I don’t see the swine industry broadly vaccinating for hepatitis E virus, but if we could implement some kind of cost-effective screening or vaccination at these upstream boar facilities, maybe we could reduce introduction of the virus into new herds.” 

This work was supported by state and federal funds appropriated to the Ohio Agricultural Research and Development Center and by the National Institute of Allergy and Infectious Diseases. 

Additional co-authors include Patricia Boley, Thamonpan Laocharoensuk, Saroj Khatiwada, Carolyn Lee, Menuka Bhandari and Juliette Hanson of Ohio State and Lindsey Moore of the College of Wooster.

This change to HIV-1 RNA, a tiny chemical modification on the adenosine building block of RNA known as m6A, is a common RNA editing process in all life forms that involves altering gene expression and protein production. The functional effect often represents a cellular solution but, in some cases, leads to disease.

By developing technological advances to observe a full length of HIV-1 RNA, researchers at The Ohio State University discovered the m6A modification occurs nearly exclusively at three specific locations on the HIV-1 RNA genome – out of the total 242 potential sites that can harbor an m6A – and these three m6As are crucial in viral replication. The finding suggested that redundancy was built into the system, and further analyses suggested that is, indeed, the case with HIV-1. 

“These sites are very important for producing virus proteins and for producing viral genomic RNA,” said senior study author Sanggu Kim, associate professor of veterinary biosciences and an investigator in the Center for Retrovirus Research at The Ohio State University. 

“An intriguing question is, why does HIV maintain multiple m6As? Our conclusion is that m6A is so important that HIV wants to have multiples to have redundancy. If it loses one or two, it’s OK. If it loses all three, it’s a problem.” 

Though any drug development associated with this work is years away, Kim said the finding suggests targeting the site-specific m6A modifications could be the basis of designing an important new treatment for HIV infection. 

The study is published today (April 11, 2024) in the journal Nature Microbiology. 

HIV-1, the most common type of the human immunodeficiency virus, attacks immune cells and uses them to make copies of itself. An estimated 1.2 million people in the United States have HIV, according to the 2023 report from the Centers for Disease Control and Prevention. 

The virus is a good example of why research on RNA modification has been getting a lot of attention in recent years, Kim said. Once thought of as the “middle guy” between DNA’s genes and life-sustaining proteins, RNA is now known to contain not just genetic information, but also to possess functional significance – in part because of the chemical modifications that accompany its messenger task. 

“Especially because HIV is an RNA virus with a very compact RNA genome, it has to encode all of the survival information within its RNA genome – it’s using not only nucleotide sequences, but all of the chemical and structural features of RNA as codes to execute its infection of host cells,” Kim said. “We know every aspect of RNA function is very important, but we don’t really know how exactly these chemical and structural modifications of RNAs regulate virus infection.” 

Though the m6A (short for N6-methyladenosine) modification was known to exist in HIV-1, previous studies had produced conflicting results about whether it helped or harmed the virus, primarily because its location was unknown and efforts to understand its effect were based on knocking out host cell genes rather than mutating the virus genome itself. 

Kim and colleagues used – and refined – a technique called nanopore direct RNA sequencing to view a full length of HIV-1’s RNA genome, which is tricky to observe because RNA is a notoriously unstable and complex molecule.   

The team first discovered the three m6A modifications and their specific locations. From there, the researchers analyzed individual RNA molecules with distinct ensembles of m6A modifications, including those with multiple m6As and those with just one of the three m6As. They found that any ensemble of m6A modifications, regardless of the number or the position of m6As, produced similar functional changes. Removal of all three, however, caused devastating effects to viruses – a dead giveaway that these m6As are redundant. 

“Until now we didn’t know which exact nucleotides are modified and how they function, and how it’s important for viruses or how it’s important for cells. Our paper addresses the keys to these important questions,” Kim said. 

“Why would HIV need all three modifications if they’re functioning in the same way?” he said. “Our study is the first to show that HIV-1 utilizes this unique, important mechanism at the RNA level for its evolutionary benefit.” 

Almost all existing HIV drugs block virus replication, but no medications inhibit viral RNA and protein production. There is more to learn about the RNA modification in HIV-1, but Kim said the work hints at the potential to develop therapies that could target these later steps.

This research was funded by the National Institutes of Health, U.S. Department of Defense, U.S. Department of Energy and the C. Glenn Barber Fund Trust.

Co-authors include Alice Baek, Ga-Eun Lee, Sarah Golconda, Anastasios Manganaris, Shuliang Chen, Nagaraja Tirumuru, Hannah Yu, Shihyoung Kim, Christopher Kimmel, Olivier Zablocki and Matthew Sullivan of Ohio State, Asif Rayhan and Balasubrahmanyam Addepalli of the University of Cincinnati, and Li Wu of the University of Iowa.

However, the study in cell cultures showed this SARS-CoV-2 variant can infect human cells that line the lower lung and engage in virus-host cell membrane fusion more efficiently, two features linked to severe disease symptoms. 

The study is published today (Jan. 8, 2024) in the journal Cell. 

The BA.2.86 variant of omicron is the ancestor of the currently dominating JN.1 and has about 60 more spike protein mutations than the original, or parent, coronavirus, including over 30 more than its close omicron relatives – the early BA.2 variant and the recently dominant XBB.1.5 variant among them. These mutations led scientists to worry that so many changes would make the variant as tough to contain as the initial omicron outbreak in 2021-22. 

“We found that, surprisingly, despite all those 60 mutations combined together, BA.2.86 is not as immune-evasive as the XBB.1.5 variant, which until recently had been dominating the pandemic for months. That’s good news,” said Shan-Lu Liu, senior author the study and a virology professor in the Department of Veterinary Biosciences at The Ohio State University. 

“But BA.2.86 appears to have increased infectivity of human lung epithelial cells compared to all omicron variants, so that’s a little worrisome. And, consistent with infectivity, it also has increased fusion activity with human lung epithelial cells,” said Liu, also a professor in the Department of Microbial Infection and Immunity. “That raises a potential concern about whether or not this virus is more pathogenic compared to recent omicron variants.” 

The published findings coincide with reports from the Centers for Disease Control and Prevention that after a brief increase in BA.2.86 infections, its derived sublineage JN.1 rapidly gained ground in the United States, responsible for an estimated 62% of COVID-19 cases as of Jan. 6. 

First detected in July in Europe and the Middle East, BA.2.86 and its sublineages have since been spreading with increasing frequency in different parts of the world. On Nov. 22, the World Health Organization classified BA.2.86 and sublineages as “variants of interest.” 

The Ohio State researchers analyzed neutralizing antibodies in blood serum samples from health care professionals who had received three monovalent vaccine doses or two monovalent vaccines followed by one bivalent vaccine booster, and from first responders who had COVID-19 infections during the wave dominated by the XBB.1.5 variant. They compared the ability of neutralizing antibodies to block infection by BA.2.86, an XBB-derived variant known as FLip, the parent virus and several omicron variants. 

Overall, antibodies produced by serum from the bivalent vaccine-dosed health care professionals were more efficient at neutralizing BA.2.86 than they were at neutralizing other omicron variants, including XBB.1.5. In contrast, the three monovalent vaccines and previous XBB.1.5 infection were barely effective in blocking infection by BA.2.86.

“People who have had a COVID-19 infection should remember that omicron variants are less virulent compared to prior variants such as delta, meaning they don’t make most people very sick,” Liu said. “If you have less severe disease, the antibodies generated by infection are low – almost 10-fold lower than vaccine-induced antibodies. That is why you cannot rely on natural infection alone for immunity. 

“While bivalent vaccine can still neutralize BA.2.86, the efficiency is clearly reduced. Therefore, it is important to get the newest booster vaccine, which is formulated with only XBB.1.5 and has been shown to be effective against BA.2.86,” Liu added. 

However, the researchers were surprised to find that a monoclonal antibody known as S309, which has been shown to inhibit almost all other major omicron variants, does not neutralize BA.2.86. Molecular modeling revealed that some of the BA.2.86 mutations in the spike protein might have changed the conformation and rendered S309 unable to neutralize the new variant, Liu said. 

Additional experiments pointed to the potential for BA.2.86 to be more likely to cause severe disease than its omicron relatives. Researchers found BA.2.86 was more efficient at infecting a cell line derived from the human lower airway epithelium in the lung. Infection of these cells is greatly facilitated through a cell surface protein, known as TMPRSS2, to promote membrane fusion, and this protein is a known contributor to SARS-CoV-2 infection and disease symptoms in the respiratory tract. 

First author Panke Qu, a graduate student in Liu’s lab, conducted the cell-culture studies using pseudoviruses – a non-infectious viral core surrounded by different SARS-CoV-2 spike proteins on the surface structured to match known variants. 

“Because we used a pseudovirus, we need to confirm these findings using the real virus,” said Liu, also associate director of Ohio State’s Center for Retrovirus Research and a program co-director of the Viruses and Emerging Pathogens Program in Ohio State’s Infectious Diseases Institute. “But from our past experience, we know that the infectivity in human epithelial cell lines provides very important information. The concern is whether or not this variant, as well as its descendants including JN.1, will have an increased tendency to infect human lung epithelial cells similar to the parental virus that launched the pandemic in 2020.” 

Liu noted that other labs have recently suggested that JN.1, one of fastest-growing descendants of BA.2.86, is much more resistant to neutralizing antibodies that are effective against BA.2.86. 

“We know that coronaviruses are prone to viral recombination, which can lead to new variants with huge numbers of mutations that could have increased immune evasion but also disease severity,” he said. “That’s why surveillance of the variants is still very important, even though we are in the end of year four of the pandemic.” 

This work was supported by anonymous donor funds, the National Institutes of Health, the National Center for Advancing Translational Sciences, the Glenn Barber Fellowship from Ohio State’s College of Veterinary Medicine, the Robert J. Anthony Fund for Cardiovascular Research, and Ohio State’s Comprehensive Cancer Center and Center for Clinical and Translational Science. 

Additional co-authors, all from Ohio State, are Kai Xu, Julia Faraone, Negin Goodarzi, Yi-Min Zheng, Claire Carlin, Joseph Bednash, Jeffrey Horowitz, Rama Mallampalli, Linda Saif, Eugene Oltz, Daniel Jones and Richard Gumina.

The scientists from The Ohio State University have reported in the Journal of Medical Entomology on the state’s first known established population of Asian longhorned ticks, and are now conducting research focused on monitoring and managing these pests.

So far, these ticks are not deemed to be a threat to human health. They tend to favor large livestock and wildlife, such as cattle and deer. Just a handful of the hundred ticks from the farm screened for infectious agents tested positive for pathogens, including one, Anaplasma phagocytophilium, that can cause disease in animals and humans. Elsewhere this tick carries another pathogen, Theileria orientalis, that affects cattle, and cases of bovine theileriosis have been reported in Ohio. 

Researchers say the tiny brown ticks – the size of a sesame seed in some life stages and pea-sized when engorged – are persistent, however: Surveillance showed they returned the following summer to the farm despite the application of pesticides in 2021. 

“They are going to spread to pretty much every part of Ohio and they are going to be a long-term management problem. There is no getting rid of them,” said Risa Pesapane, senior author of the paper and an assistant professor of veterinary preventive medicine at Ohio State. 

“The good news about the ticks, though, is that most tick control agents that we currently have seem to kill them. Still, managing them is not easy because of how numerous they are and how easily they can come back.” 

Asian longhorned ticks originate from East Asia and were first detected in the United States in New Jersey in 2017. When Pesapane joined Ohio State in 2019 as a tick-borne disease ecologist, the ticks were reported in West Virginia – meaning it was only a matter of time before they crossed the river into Ohio, she said. 

She found the first of these ticks in Ohio, on a stray dog in Gallia County in 2020, and another was collected from a cow in Jackson County in June 2021. And then a farmer from Monroe County called Ohio State later that summer to report three of his 18 cattle, heavily infested with ticks, had died.

“One of those was a healthy male bull, about 5 years old. Enormous. To have been taken down by exsanguination by ticks, you can imagine that was tens of thousands of ticks on one animal,” said Pesapane, who also has a faculty appointment in Ohio State’s School of Environment and Natural Resources. 

Pesapane and colleagues collected almost 10,000 ticks within about 90 minutes on the farm, leading her to speculate that there were more than 1 million of them in the roughly 25-acre pasture. 

Asian longhorned ticks’ secret colonization weapon is the ability to reproduce asexually, with each female laying up to 2,000 eggs at a time – and all 2,000 of those female offspring able to do the same. 

“There are no other ticks in North America that do that. So they can just march on, with exponential growth, without any limitation of having to find a mate,” Pesapane said. “Where the habitat is ideal, and anecdotally it seems that unmowed pastures are an ideal location, there’s little stopping them from generating these huge numbers.”

Because of their ability to hide in vegetation, Asian longhorned ticks also can escape pesticides that kill only when coming into direct contact with a pest. 

“It would be wisest to target them early in the season when adults become active, before they lay eggs, because then you would limit how many will hatch and reproduce in subsequent years. But for a variety of reasons, I tell people you cannot spray your way out of an Asian longhorned tick infestation – it will require an integrated approach,” Pesapane said. 

She and colleagues are working as rapidly as they can at filling in knowledge gaps about these invaders and developing training materials and policy recommendations for affected industries. As one example, Pesapane said, tick inspections of livestock could provide a window for application of an antiparasitic agent to eliminate the risk of transporting the exotic arachnids across multiple state lines. 

Ohioans are encouraged to help with research efforts: People who think they’ve spotted an Asian longhorned tick can email ticks@osu.edu for instructions on how to collect the specimen and send it to Ohio State scientists as part of ongoing surveillance. To date, the lab has received Asian longhorned ticks from residents of 11 Ohio counties. 

More information about spotting Asian longhorned ticks and preventing tick exposure is available on Ohio State’s Bite Site hosted by the College of Food, Agricultural, and Environmental Sciences Knowledge Exchange. 

This work was supported by the U.S. Department of Agriculture. Co-authors on the study were Andreas Eleftheriou, Julia Beckett and Ningzhu Bai, all of Ohio State.

The study builds upon previous studies that involved inserting a highly stable segment of the coronavirus spike protein into the measles vaccine or mumps vaccine. 

In a paper publishing this week in Proceedings of the National Academy of Sciences, scientists at The Ohio State University report on a new MMS vaccine candidate – for Measles, Mumps and SARS-CoV-2 – delivered via the nose that provides broad and long-lasting protection against COVID-19 infection. 

In two rodent models, the intranasal vaccine triggered a strong neutralizing antibody response plus protection in mucosal areas lining the nose and lungs, and prevented disease symptoms such as weight loss and tissue damage. 

Experiments suggested the lifelong immunity to measles and mumps provided by the measles-mumps-rubella (MMR) vaccine would likely translate into prolonged protection against COVID-19 in people vaccinated with the MMS: In hamsters, antibodies against SARS-CoV-2 induced by the MMS vaccine lasted at least four months without any sign of decline. 

The trivalent vaccine developed in this study protected against the ancestral SARS-CoV-2 virus and two variants: delta, which was associated with more severe disease, and omicron BA.1, which spread much faster. 

“The beauty here is we already know the MMR is used in children, so we’re building on a 50-year safety record,” said Jianrong Li, senior author of the study and a professor of virology in Ohio State’s Department of Veterinary Biosciences and Infectious Diseases Institute. “We inserted three different spikes that allow broad neutralizing antibodies to protect against the different variants of concern of SARS-CoV-2. It’s quite promising, and would be a fantastic new type of vaccine to prevent COVID-19.” 

In addition to the promising platform and the bonus mucosal protection that comes with intranasal delivery, the technique used to create the coronavirus antigens that stimulate immunity contributes to the vaccine candidate’s effectiveness, Li said. 

Researchers used a prefusion version of each variant’s spike protein – the shape it is in on the viral surface before the virus infects a cell. The protein, called preS-6P, was locked into this form by changing six of its amino acids to prolines. In a previous study published in PNAS, Li’s lab found that preS-6P is much more effective in inducing immune responses than preS-2P, a spike locked by two prolines. Currently, all FDA-approved COVID-19 vaccines use the preS-2P. 

“The efficacy and longevity of COVID-19 vaccines will be much higher if preS-6P is used,” Li said. 

Using one measles strain and two mumps strains, the team inserted the antigens in the same location in each platform virus’s genome to enable expression of the optimal amount of spike protein and robust replication of the platform viruses – both actions needed to trigger a strong immune response. 

Tests in mice used to model an immune response showed intranasal delivery of the MMS vaccine generated neutralizing antibodies in the bloodstream against the three variants as well as specialized antibodies called IgA on mucosal surfaces of the airways and tissue-resident T cells in lungs that help boost clearance of viral particles. 

“These are two main advantages of intranasal vaccines: generating IgA in the nose and lung to prevent the virus from traveling to other organs or the blood, and producing tissue-resident T cells in the lung to rapidly mount an effective immune response to previously encountered pathogens, in this case SARS-CoV-2,” Li said. “The mRNA vaccines injected intramuscularly primarily generate antibodies in the bloodstream, but do not produce these types of added protection for the lungs.” 

Experiments comparing the trivalent vaccine candidate to a vaccine against a single omicron variant showed the single-variant version was not able to produce antibodies that neutralize other coronavirus strains. 

“With the insertion of the three spike proteins, the MMS produces antibodies that neutralize other strains – that supports the concept of using a trivalent vaccine to produce broader immunity against different strains,” said co-first author Jiayu Xu, a graduate student in Li’s lab. 

Immunization of golden Syrian hamsters with the trivalent vaccine produced similarly high levels of neutralizing antibodies as in the mice. When the hamsters were infected with the parental, delta and omicron strains of the virus, multiple measures compared to controls showed they were protected from getting sick: a lack of clinical symptoms, only minor changes to tissue in the airways and undetectable levels of viral particles present in the lungs. 

“The MMS platform is also readily and rapidly adaptable to new variants such as XBB.1.16 and EG.5, which are currently circulating in the human population,” said co-first author Yuexiu Zhang, a graduate student in Li’s lab.  

Li said there are additional options to consider, including adding rubella back into the platform and, potentially, inserting more than three prefusion spike proteins or other coronavirus proteins into the vaccine to further broaden protective immunity. 

“We envision incorporating it into a routine immunization program for children and to provide long-term immunity against COVID-19 for adults,” he said. 

Ohio State has filed a patent application on measles virus-based SARS-CoV-2 vaccine candidates. Ohio State and Nationwide Children’s Hospital also filed a patent application on mumps virus-based SARS-CoV-2 vaccine candidates.

The study was supported by grants from the National Institutes of Health, the National Cancer Institute and Ohio State’s College of Veterinary Medicine.

Additional co-authors include Panke Qu, Mohamed Shamseldin, Sung Yoo, Jesse Hall, John Evans, Mostafa Eltobgy, Mijia Lu, Michelle Chamblee, Xueya Liang, Amal Amer, Jacob Yount, Prosper Boyaka, Shan-Lu Liu and Purnima Dubey of Ohio State; Mark Peeples, Jack Misny, Ilada Thongpan and Mahesh KC of the Abigail Wexner Research Institute at Nationwide Children’s Hospital; and Chengjin Ye and Luis Martinez-Sobrido of the Texas Biomedical Research Institute. Jason McLellan of the University of Texas at Austin discovered the preS-6P used in this study.

Researchers surveyed new owners of dogs adopted from five Ohio shelters at four time points after the pet went home – seven, 30, 90 and 180 days later – using a research tool called the Canine Behavioral Assessment & Research Questionnaire (C-BARQ). 

Results showed the dogs had a high prevalence for aggression toward strangers, their owners and other dogs that changed in a variety of ways over time, and separation-related behavior problems decreased at the six-month time point. By the study’s end, 93.7% of owners rated their dog’s overall behavior as excellent or good and 100% reported their pet had adjusted to the new home extremely or moderately well. 

“We really got to see where in the timeline the pet’s behavior may change or may not change, and that’s really the key,” said lead study author Kyle Bohland, assistant professor-clinical of behavioral medicine in the Department of Veterinary Clinical Sciences at The Ohio State University.

“The shelter system touches many lives, both humans and pets,” said Bohland, whose practice centers on companion animal behavior. “And so it’s important for us to be able to counsel owners on what may or may not change in the future so they can be better prepared to handle those consequences and then, hopefully, keep dogs in homes.” 

The study was published recently in the journal PLOS ONE.

Statistics show that about 2 million dogs are adopted from U.S. shelters every year. 

An internet search about adopting shelter dogs will return results suggesting these pets’ behavior will change three days, three weeks and three months after joining a household. Three days makes sense, Bohland said, because the stress hormone cortisol is known to spike in dogs when they enter a new environment, and then returns to normal in a few days. 

“But the three-week and three-month rule really had no scientific basis,” he said. Previous studies on adopted shelter dogs took a piecemeal approach, testing one point in time in the new home or using unvalidated surveys. 

So Bohland and his colleagues set out to take a longer look at dog behavior changes in their forever homes using C-BARQ, which provides a standardized way to assess an individual dog’s behavior. They recruited participants who adopted dogs from four shelters in the Columbus area and one in Cleveland. 

Ninety-nine new adopters completed the first survey a week after taking their dogs home, and repeat surveys were emailed to those participants one, three and six months after the adoption. Sixty-two owners answered all four surveys. 

The survey contained 42 questions asking participants to use a 0-4 scale to rate excitability; aggression directed toward strangers, owners and either familiar or unfamiliar dogs; fear; touch sensitivity; separation-related behavior; attachment and attention seeking; chasing; and energy level. Owners were also asked at each time point to rate their overall satisfaction with the dog’s behavior and document any changes in their household.

Among behaviors that changed, results showed that a few increased at all time points: stranger-directed aggression, chasing behavior and training difficulty. Increases in excitability and touch sensitivity were reported at 90 and 180 days. 

Two characteristics, separation-related behaviors and attachment and attention-seeking, decreased by the six-month mark.

Of behaviors that didn’t change, a few still stood out for their high prevalence at various time points: dog-direction aggression (75%), familiar dog aggression (37.8%), and owner-directed aggression (32.3%).

“The biggest thing that stuck out to me was that we’ve got a lot of aggression among dogs in our community. That definitely concerns me from a public health standpoint and from a human mental health standpoint, because we’ve got a lot of dogs that are struggling – and that has human implications,” Bohland said. 

“And the other big piece was that despite that, people were pretty darn happy with their dogs,” he said. “This combination of findings is a reminder that just about everybody has, on some level, dealt with unpredictable behavior problems, illnesses and the quirks of animal aging – and we still love our dogs. Overall, this really speaks to the bond people have with their pets.” 

Statistical analysis suggested that in some cases, a dog’s size, age or sex was associated with specific behaviors. Dogs treated in the shelter with anti-anxiety medications were more apt to show aggression toward strangers and touch sensitivity after adoption – likely because those pets were more difficult to handle from the beginning, not because the medications made it worse. Researchers deliberately left breed out of the analysis – most dogs were described as mixed breeds, and all were neutered. 

Based on what the researchers see in their practice, some behavior changes make sense: Stranger aggression may increase as dogs feel more protective of their new environment, and separation anxiety logically lowers as dogs settle into a household routine. Touch sensitivity might go unnoticed for months – until that first nail trim. 

Seven adopted dogs were returned to the shelter during the study, resulting in a return rate of 7.1%. The national average return rate is about 15%. 

“The bottom line is we don’t want to see dogs coming back to shelters,” Bohland said. “A lot of what we study comes from clients having questions. So my hope is that in the long term, this can help shelter employees and veterinarians target interventions that will help keep more dogs in their homes.” 

This work was funded by a grant from Ohio State’s College of Veterinary Medicine. Co-authors were M. Leanne Lilly, Andréa Arruda and Jeanette O’Quin of Ohio State, and Meghan Herron of Gigi’s Shelter for Dogs.

Scientists collected 1,522 nasal swabs from free-ranging deer in 83 of the state’s 88 counties between November 2021 and March 2022. More than 10% of the samples were positive for the SARS-CoV-2 virus, and at least one positive case was found in 59% of the counties in which testing took place. 

Genomic analysis showed that at least 30 infections in deer had been introduced by humans – a figure that surprised the research team. 

“We generally talk about interspecies transmission as a rare event, but this wasn’t a huge sampling, and we’re able to document 30 spillovers. It seems to be moving between people and animals quite easily,” said Andrew Bowman, associate professor of veterinary preventive medicine at The Ohio State University and co-senior author of the study. 

“And the evidence is growing that humans can get it from deer – which isn’t radically surprising. It’s probably not a one-way pipeline.” 

The combined findings suggest that the white-tailed deer species is a reservoir for SARS-CoV-2 that enables continuing mutation, and that the virus’s circulation in deer could lead to its spread to other wildlife and livestock. 

The study is published today in Nature Communications. 

Bowman and colleagues previously reported detection of SARS-CoV-2 infections in white-tailed deer in nine Ohio locations in December 2021, and are continuing to monitor deer for infection by more recent variants. 

“We expanded across Ohio to see if this was a localized problem – and we find it in lots of places, so it’s not just a localized event,” Bowman said. “Some of the thought back then was that maybe it’s just in urban deer because they’re in closer contact with people. But in rural parts of the state, we’re finding plenty of positive deer.”

Beyond the detection of active infections, researchers also found through blood samples containing antibodies – indicating previous exposure to the virus – that an estimated 23.5% of deer in Ohio had been infected at one time or another. 

The 80 whole-genome sequences obtained from the collected samples were represented groups of viral variants: the highly contagious delta variant, the predominant human strain in the United States in the early fall of 2021 that accounted for almost 90% of the sequences, and alpha, the first named variant of concern that had circulated in humans in the spring of 2021. 

The analysis revealed that the genetic composition of delta variants in deer matched dominant lineages found in humans at the time, pointing to the spillover events, and that deer-to-deer transmission followed in clusters, some spanning multiple counties. 

“There’s probably a timing component to what we found – we were near the end of a delta peak in humans, and then we see a lot of delta in deer,” Bowman said. “But we were well past the last alpha detection in humans. So the idea that deer are holding onto lineages that have since gone extinct in humans is something we were worried about.” 

The study did suggest that COVID-19 vaccination is likely to help protect people against severe disease in the event of a spillover back to humans. An analysis of the effects of deer variants on Siberian hamsters, an animal model for SARS-CoV-2 studies, showed that vaccinated hamsters did not get as sick from infection as unvaccinated animals.

That said, the variants circulating in deer are expected to continue to change. An investigation of the mutations found in the samples provided evidence of more rapid evolution of both alpha and delta variants in deer compared to humans. 

“Not only are deer getting infected with and maintaining SARS-CoV-2, but the rate of change is accelerated in deer – potentially away from what has infected humans,” Bowman said. 

How the virus is transmitted from humans to white-tailed deer remains a mystery. And so far, even with about 30 million free-ranging deer in the U.S., no substantial outbreaks of deer-origin strains have occurred in humans. 

Circulation among animals, however, remains highly likely. Bowman noted that about 70% of free-ranging deer in Ohio have not been infected or exposed to the virus, “so that’s a large body of naive animals that the virus could spread through rather uninhibited.” 

“Having that animal host in play creates things we need to watch out for,” he said. “If this trajectory continues for years and we have a virus that becomes deer-adapted, then does that become the pathway into other animal hosts, wildlife or domestic? We just don’t know.”

This work was supported by the National Institute of Allergy and Infectious Diseases and Ohio State’s Infectious Diseases Institute.

Martha Nelson of the National Library of Medicine was co-corresponding author of the study. Ohio State co-authors Dillon McBride, Steven Overend, Devra Huey, Amanda Williams, Seth Faith and Jacqueline Nolting worked on the study with co-authors from St. Jude Children’s Research Hospital; the University of California, Los Angeles; the National Research Centre in Giza, Egypt; PathAI Diagnostics; the Ohio Department of Natural Resources; the U.S. Department of Agriculture; Columbus and Franklin County Metroparks; and the Rega Institute for Medical Research in Belgium.

Rather than let the next outbreak take the world by surprise, two virologists say in a Science Perspective article published today (March 10, 2023) that the scientific community should invest in a four-part research framework to proactively identify animal viruses that might infect humans.

“A lot of financial investment has gone into sequencing viruses in nature and thinking that from sequence alone we’ll be able to predict the next pandemic virus. And I think that’s just a fallacy,” said Cody Warren, assistant professor of veterinary biosciences at The Ohio State University and co-lead author of the article.

“Experimental studies of animal viruses are going to be invaluable,” he said. “By measuring properties in them that are consistent with human infection, we can better identify those viruses that pose the greatest risk for zoonosis and then study them further. I think that’s a realistic way of looking at things that should also be considered.” 

Warren co-authored the opinion piece with Sara Sawyer, professor of molecular, cellular and developmental biology at the University of Colorado Boulder. 

One key message Warren and Sawyer want to get across is that knowing an animal virus can attach to a human cell receptor doesn’t paint the whole picture of its zoonotic potential. 

They propose a series of experiments to assess an animal virus’s potential to infect a human: If it is found to enter human cells, can it use those host cells to make copies of itself and multiply? After viral particles are produced, can they get past human innate immunity? And have human immune systems ever been exposed to another virus from the same family? 

Answering these questions could enable scientists to put a pre-zoonotic candidate virus “on the shelf” for further research – perhaps developing a quick way to diagnose the virus in humans if an unattributable illness surfaces and testing existing antivirals as possible treatments, Warren said. 

“Where it becomes difficult is that there may be many animal viruses out there with signatures of human compatibility,” he said. “So which ones do you pick and choose to prioritize for further study? That’s something that needs to be carefully considered.” 

A decent starting point, he and Sawyer suggest, would be operating on the assumption that viruses with the most risk to humans come from “repeat offender” viral families currently infecting mammals and birds. Those include coronaviruses, orthomyxoviruses (influenza) and filoviruses (causing hemorrhagic diseases like Ebola and Marburg). In 2018, the Bombali virus – a new ebolavirus – was detected in bats in Sierra Leone, but its potential to infect humans remains unknown. 

And then there are arteriviruses, such as the simian hemorrhagic fever virus that exists in wild African monkeys, which Sawyer and Warren recently determined has decent potential to spill over to humans because it can replicate in human cells and subvert immune cells’ ability to fight back. 

The 2020 worldwide lockdown to prevent the spread of COVID-19 is still a fresh and painful memory, but Warren notes that the terrible outcomes of the emergence of SARS-CoV-2 could have been much worse. The availability of vaccines within a year of that lockdown was possible only because scientists had spent decades studying coronaviruses and knew how to attack them. 

“So if we invest in studying animal viruses early and understand their biology in more detail, then in the case that they were to emerge in humans later, we’d be better poised to combat them,” Warren said. 

“We are continually going to be exposed to the viruses of animals. Things are never going to change if we stay on the same trajectory,” he said. “And if we stay complacent and only study those animal viruses after they jump into humans, we’re constantly going to be working backwards. We’ll always be behind.”

Jianrong Li, professor of virology in the College of Veterinary Medicine Department of Veterinary Biosciences, and Peixuan Guo, holder of the Sylvan G. Frank Endowed Chair in Pharmaceutics and Drug Delivery in the College of Pharmacy, are among the 169 academic inventors to be named National Academy of Inventors (NAI) fellows this year. They are the 15th and 16th Ohio State inventors to be chosen as NAI fellows. 

In 2021, Guo was named Ohio State’s Innovator of the Year, and Li, along with colleague Stefan Niewiesk, was among the finalists for the award.

Jianrong Li 

Li has led the development of a number of vaccine candidates for respiratory viruses and enteric illnesses – those that affect the respiratory and gastrointestinal systems. His lab worked swiftly to develop vaccines against the SARS-CoV-2 virus that causes COVID-19 in response to the pandemic, developing separate candidates using the measles and mumps viruses as platforms for a COVID-19 vaccine. 

“The idea is that because children have been safely vaccinated for measles and mumps for decades, we could incorporate this vaccine into the MMR vaccine to prevent four diseases simultaneously: measles, mumps, rubella and COVID-19,” Li said. 

These platforms also enable delivering a “needle-free” vaccine through the nose, which generates both a system-wide antibody response – the same response as that produced by an injection in the arm – as well as a mucosal immune response in the linings of airways, where viral particles attach to host cells. All currently approved COVID-19 vaccines are delivered intramuscularly but are unable to induce sufficient mucosal immunity, the body’s first line of defense against respiratory viruses. The White House’s effort to develop the next generation of COVID-19 vaccines includes an intranasal vaccine approach. 

In each candidate, the team’s use of specific snippets of the “prefusion” version of the SARS-CoV-2 spike protein – the shape the protein is in before it infects a cell – and strategic insertion of those genes into modified measles and mumps virus genomes produced strong neutralizing antibody, mucosal antibodies and T cell immune responses and protected against lung damage in animals. 

The SARS-CoV-2 vaccine candidates and a third vaccine candidate against respiratory syncytial virus (RSV), the No. 1 cause of human respiratory infections that can also cause serious illness in infants and the elderly, have been licensed to an India-based vaccine and pharmaceutical company. 

Vaccines developed in Li’s lab have also been commercialized for use in protecting piglets from two deadly viruses, porcine epidemic diarrhea virus and porcine deltacoronavirus. Previous viral outbreaks have caused significant economic losses for the pork industry in the United States and worldwide. 

A member of Ohio State’s Infectious Diseases Institute, Li has been awarded eight patents and has eight pending patent applications, and has entered into a total of eight licensing agreements with companies that are commercializing vaccine products developed in his lab. 

Li, who joined Ohio State’s faculty in 2008, attributed his lab’s success to Ohio State’s creative and collaborative environment and support from his college and department. 

“I am humbled and truly honored that my peers have considered me for this award,” he said. “This honor really belongs to the postdoctoral fellows, graduate students and research assistants in my laboratory. In the past 14 years at Ohio State, I have had the privilege to work with them to generate innovative ideas, make new scientific discoveries, and translate basic virology research into the development of novel vaccines against viral pathogens that threaten human and animal health.” 

Peixuan Guo 

The many RNA-related discoveries attributed to Guo, also director of the College of Pharmacy Center for RNA Nanotechnology and Nanomedicine, date back to his days as a student in the 1980s, when he determined that short segments of noncoding RNA not only existed, but had important functions in cells. In findings published in Science, he called them “small RNA” or sRNA. Among those he proposed at the time were the microRNAs that have been found since to have a powerful influence over genes’ protein-building functions. 

Guo’s discoveries involve some of the most intricate cellular processes – many of which have opened the door to the development of therapies, particularly for several types of cancers. He is a member of the Ohio State University Comprehensive Cancer Center Translational Therapeutics Program. 

Guo was a pioneer in the field of RNA nanotechnology, first proving RNA strands could self-assemble into a nanoparticle, and since advancing understanding of RNA particles and properties that make them useful vehicles for targeted delivery of anticancer drugs. In 2014, he proposed that RNA therapeutics would be the third milestone in pharmaceutical drug development. 

In 2020, his lab showed in animal studies that RNA nanoparticles have rubbery and amoeba properties that help explain why they target cancer tumors so efficiently and how they keep their toxicity low – by quickly leaving the body. 

He reported in 2021 on a promising liver cancer therapy: an RNA nanoparticle designed to carry both a chemotherapy drug and a molecule to prevent the pumping of the drug out of liver cells. In animal studies, these nanoparticles targeted tumor cells effectively and inhibited tumor growth. 

“RNA can be therapeutic, but it has taken some time for the field to get to pharmaceutical development,” he said. “The mRNA vaccines have opened up the public’s understanding of RNA. At one time, people didn’t believe it was possible – but now we know it is.” 

Guo also points to his first construction of a viral DNA packaging motor. He discovered the third type of biomotors that use a revolving mechanism without rotation to transport the lengthy dsDNA genome. A biomotor is a biological machine that drives movement to help with the energy-related transport of proteins and other molecules – key processes behind such activities as walking and breathing, because transport is essential to the completion of numerous cellular functions. The use of the revolution mechanism without the use of rotation avoids the coiling or tangling during the transportation of the lengthy genome. 

The finding eventually led to applications of this motor to perform “single pore sensing,” including the detection of individual peptides – short chains of amino acids – to advance cancer diagnostics.  

Guo, too, took on new projects at the start of the pandemic, including development of an at-home diagnostic test for COVID-19 infection. An invention of his dating back more than 30 years made an indispensable contribution to the mRNA vaccines against SARS-CoV-2: a novel way of producing a molecule of vaccinia virus mRNA capping enzyme that prevents mRNA from degradation and ensures proper translation of mRNA protein-manufacturing instructions. 

Guo has been a member of Ohio State’s faculty since 2015. He has filed 70 patent applications, with 13 granted and others pending. Most of these technologies have been licensed for commercialization. 

“Of all the recognitions I have received, this is the highest honor in the field of innovation,” Guo said. “I am happy to be recognized as an NAI fellow.” 

The NAI Fellows Program highlights academic inventors who have demonstrated a spirit of innovation in creating or facilitating outstanding inventions that have made a tangible impact on the quality of life, economic development and the welfare of society. 

“This year’s class of NAI fellows represents a truly outstanding caliber of innovators,” said Paul R. Sanberg, president of the NAI. “The breadth and scope of their inventions is truly staggering. I am excited to see their creativity continue to define a new era of science and technology in the global innovation ecosystem.” 

The 2022 class of fellows will be inducted at the annual meeting of the National Academy of Inventors in June 2023 in Washington, D.C.

Scientists tested neutralizing antibodies in blood serum samples from vaccinated and once-boosted or recently infected health care professionals against several subvariants in circulation. Three subvariants stood out for their resistance to the antibody immune response: BQ.1, BQ.1.1 and BA.2.75.2.

BQ.1 and BQ.1.1 are subvariants of the BA.4/5 omicron variants that have been dominating the last few months in the U.S., and each now accounts for about a quarter of current infections, according to the Centers for Disease Control and Prevention (CDC). BA.2.75.2, a mutant of the BA.2 omicron variant, was the best of all variants tested at evading neutralizing antibodies, but currently accounts for only a very small proportion of reported illnesses in the United States.

“In general, the subvariants BQ.1 and BQ.1.1 are much better compared to prior variants at evading the booster-mediated antibody response – the neutralizing antibody titers are clearly much lower. And those two variants are becoming dominant,” said Shan-Lu Liu, senior author the study and a virology professor in the Department of Veterinary Biosciences at The Ohio State University.

“It is important to be aware while traveling and gathering over the holidays that SARS-CoV-2 variants are continuing to evolve,” said Liu, also a professor in the Department of Microbial Infection and Immunity. “If you were vaccinated with the first booster more than six months ago, you might consider getting a second booster because the antibodies from a single booster are now likely too low to be protective.”

The study is published today in the journal Cell Host & Microbe. 

Though the third booster, an updated bivalent formulation, is now being administered to those who are eligible, over 48% of the total booster-eligible population has not yet received a booster dose, according to CDC data. Liu’s lab has previously shown that a COVID-19 booster shot provided strong and broad antibody protection against a range of omicron sublineage variants, and that a second booster restored waning antibody counts to protective levels.

For the current study, serum samples came from health care professionals who had received two mRNA vaccine doses and one booster shot or who had been infected during an early or later omicron wave.

Results showed about a 20-fold decrease in vaccine and single booster-generated antibodies that could neutralize BQ.1 and BQ.1.1 compared to neutralizing antibodies against the original, or parent, SARS-CoV-2 virus. Similarly, neutralizing antibody levels, or titers, generated by infection during the BA.1 omicron wave were significantly lower against the BQ subvariants than against the parental virus, and antibody titers against the BQ subvariants generated by infection during the BA.4/5 wave did not reach the level of detection.

“Our results suggest that you cannot count on natural infection to protect against the currently circulating omicron subvariants,” said Liu, also associate director of Ohio State’s Center for Retrovirus Research and a program co-director of the Viruses and Emerging Pathogens Program in Ohio State’s Infectious Diseases Institute.

Liu and colleagues conducted cell-culture studies using pseudoviruses – a non-infectious viral core surrounded by different SARS-CoV-2 spike proteins on the surface structured to match known variants. The method used to detect neutralizing antibodies in the blood samples accounts for the varying levels of antibodies produced by individuals. 

In this study, Ohio State collaborator and co-author Kai Xu created structural models of individual amino acids changed by the newest mutations on the spike protein, identifying a few key molecules that the subvariants have rearranged so they can block antibodies from binding to the viral particles. This modeling showed that one of these amino acids, called N460K, also enables the BQ.1 and BQ.1.1 particles to enter host cells more efficiently and force host cells to fuse together, a step in viral infection that can increase pathogenesis – which contributes to disease onset, progression to more severe symptoms, and disease transmissibility.

“From my perspective, this is cause for concern because the original omicron variant was not very pathogenic – it did not cause much cell fusion at all,” Liu said. “But we now see a trend with these new subvariants of increased cell fusion, and that trend puts the virus in a better position to cause infection and pathogenesis.”

Other labs’ studies of the bivalent booster’s neutralizing antibody production suggest the updated booster would offer protection against the newer omicron subvariants, Liu said.

But Liu has submitted a paper for publication about another subvariant called XBB, a recombinant variant created by the exchange of genetic material between two BA.2 omicron subvariants, that shows what he calls “extraordinary” resistance to neutralizing antibodies produced by vaccination and previous infection.

“This virus can do unexpected things,” he said. “We still have to do a good job of surveillance and keep our eye on these emerging variants of concern.”

This work was supported by anonymous donor funds, grants from the National Institutes of Health, the National Center for Advancing Translational Sciences, the Glenn Barber Fellowship from Ohio State’s College of Veterinary Medicine, the National Cancer Institute, the Robert J. Anthony Fund for Cardiovascular Research, and Ohio State’s Comprehensive Cancer Center, Office of Health Sciences, and Center for Clinical and Translational Science.

Additional co-authors, all from Ohio State, are Panke Qu, John Evans, Julia Faraone, Yi-Min Zheng, Claire Carlin, Mirela Anghelina, Patrick Stevens, Soledad Fernandez, Daniel Jones, Gerard Lozanski, Ashish Panchal, Linda Saif, Eugene Oltz and Richard Gumina.

The College of Medicine, College of Nursing, College of Optometry, College of Public Health and College of Veterinary Medicine were recognized for demonstrating an outstanding commitment to diversity and inclusion. Ohio State is the only academic institution in the country to have five colleges honored with the Health Professions HEED award.

“This recognition is meaningful and reflects the outstanding work of these five colleges to further diversity and inclusion,” said Melissa Gilliam, executive vice president and provost at Ohio State. “By fostering a culture of inclusive excellence, the entire university community benefits through a greater sense of belonging.”

INSIGHT Into Diversity magazine evaluated academic institutions on leadership support for diversity, recruitment and retention of students and employees, and other aspects of campus diversity and inclusion. Ohio State diversity and inclusion efforts include:

College of Medicine: The College of Medicine has adopted curriculum and training procedures to explore racial disparities in medicine with the goals of dispelling myths and becoming aware of and rectifying any subconscious bias. Coupled with the classroom education are opportunities to apply learnings in the clinical skills lab through the use of manikins of color and melanated suture pads.

College of Nursing: The Summer Institute for Future Nurses introduces diverse groups of high school students from around the country to nursing. Offered in a hybrid format, students participated in mock clinical skills training sessions on campus and virtual presentations. More than 20 previous attendees of the Summer Institute have graduated from the College of Nursing with a Bachelor of Science in Nursing degree. 

College of Optometry: BuckEye Launch is an immersive five-day program for first-year students that provides strategies for academic success and creates a sense of community among students, faculty and staff. Major themes include effective learning and test taking strategies, self-care and personal wellness, and time management.

College of Public Health: The College of Public Health dean’s administrative leadership team and extended staff applied for and were chosen to be part of the inaugural class of Ohio State’s Office of Diversity and Inclusion’s new Inclusive Excellence Team Certification Program. The nine-month intensive learning and project development opportunity is designed to ensure that each participating faculty and staff member receives the same inclusive excellence education and tools to build a diverse, equitable and inclusive workplace. It includes a capstone project designed to advance the college’s goals around diversity, equity and inclusive excellence in the long term.

College of Veterinary Medicine: Diversity, equity, inclusion and belonging (DEIB) is incorporated as a foundational principle in each of the six goal areas of the College of Veterinary Medicine’s Be The Model® strategic plan. The formation of a new student leadership group, Student Leaders for Inclusive Excellence, is one example of many initiatives designed to support DEIB in the college community. Comprised of student leaders representing the college’s affinity groups and student chapters supportive of DEIB, this group provides mentorship and support to student affinity groups and student organizations that are supportive of diverse populations. The goal is for all student organizations to work together to create a sense of belonging with one another and to develop leadership skills as they plan DEIB educational events for the entire college community. 

Ohio State will be featured in the December 2022 issue of INSIGHT Into Diversity magazine.

In a perspective article published this week in Proceedings of the National Academy of Sciences, the 14 experts cite virus pandemics dating from 1918 to the COVID-19 crisis as examples of how “the world has largely failed to meet the challenge to be better prepared to prevent or respond to the next outbreak.” 

Future outbreaks are inevitable. The team says the best way to lower chances for widespread disease outbreaks and pandemics and improve prospects for mounting a rapid response is to adopt a One Health approach to diminish these threats – working across disciplines and administrative barriers at all levels to understand and address links between animal and human health and the environment. 

Ohio State University virologist and immunologist Linda Saif, co-lead author of the paper, has been sounding the alarm about viruses that affect food animals, wildlife and humans for decades – and provided expertise very early in the pandemic about the dangers of SARS-CoV-2. In 1995, her lab was the first to document a coronavirus’s leap from wild animals to cattle and from cattle to poultry. The recognition of interspecies transmission of coronaviruses from deer to cattle was prescient: In 2021, Saif was part of an Ohio State research team that showed that SARS-CoV-2 could spill over to deer. All these years later, the scientific community is still learning about how the wily pathogens work.

“SARS-CoV-2 taught us that viruses do not respect borders, walls, demographics or politics – nor do they respect species barriers,” said Saif, a Distinguished University Professor in Ohio State’s Center for Food Animal Health with faculty appointments in the departments of animal sciences and veterinary preventive medicine.

“Emerging and re-emerging RNA viruses – including coronaviruses – are a major cause of the transmission of disease from animals to humans and back again to animals, and that spillover between species enables viruses to establish new hosts in which they can mutate and persist. The  most effective way to fight back is to work as a global community and apply One Health practices for prevention and preparedness.” 

The researchers from the United States, Africa, Asia, Australia and Europe convened in 2021 as the Independent Task Force on COVID-19 and Other Pandemics: Origins, Prevention and Response. The task force was chaired by Gerald Keusch of the National Emerging Infectious Diseases Laboratory and Center for Emerging Infectious Diseases Policy and Research at Boston University, who is co-lead author of the PNAS article. 

Recommendations are based on their findings from an expansive review of major RNA virus outbreaks over the last 50 years and research results before and during the COVID-19 pandemic. They paid particular attention to identifying places and times when targeted interventions in the past could have blocked cross-species transmission to inform their proposed solutions for the future. 

Evidence strongly suggests that the two SARS coronavirus outbreaks, in 2003 and 2019, can be traced to coronaviruses in bats that most likely spilled over to intermediate animal hosts in wildlife farms or markets before infecting people – in the case of COVID-19, at the Huanan Seafood Market in Wuhan, China. The task force found the risk for a pandemic increases when people and animals interact closely in altered settings driven by land use and climate change, environmental degradation, the wildlife trade, population growth and economic pressure.

Addressing the  risk factors of these types of conditions is one focus of the group’s recommendations, which include:

• Conducting early-warning surveillance in locations where people, wildlife and domestic animals intermingle to detect high-threat potential zoonotic pathogens and inform development of potential broad spectrum vaccines and therapeutics.
• Investing in research and development of diagnostics, antivirals and vaccines for priority pathogens, and streamlining avenues to enable rapid clinical testing and manufacturing of medical countermeasures.
• Reducing drivers for spillover risk and spread, in part by minimizing high-risk human-wildlife contact, on the front lines of disease emergence from the community to country level.
• Countering misinformation and disinformation about the prevention and control of emerging diseases based on research focused on fostering public trust in and understanding of science and expert advice, and providing trusted resources and outlets for accurate information.
• Establishing an inclusive, transparent One Health governance framework at all levels for pandemic preparedness and response, and providing stable funding for all related global efforts.

“It’s not an overstatement to say that this requires worldwide collaboration and coordination to come up with measures enabling us to predict, prevent, mitigate and control future pandemics,” Saif said. “We know where opportunities have been missed in the past. We know what research questions need to be answered most urgently. We just have to seize the opportunity and have the resolve to act on what we know to improve human, animal and our ecosystem health.”

This look toward a better future comes at a time when SARS-CoV-2 is still causing infections worldwide and still has potential to circulate as viral variants that would pose new threats to human health and, as spillovers continue, potentially to animal health, the researchers noted. 

“The time to energize these processes is now,” the task force concludes, “when the tragedy of COVID-19 continues to confront the public and politicians.”

Additional task force co-authors include John Amuasi of Kwame Nkrumah University of Science and Technology, Ghana; Danielle Anderson of the The Peter Doherty Institute for Infection and Immunity, University of Melbourne, Australia; Peter Daszak, and Su Yadana of EcoHealth Alliance, New York City; Isabella Eckerle of University Hospital of Geneva, Switzerland; Marion Koopmans of  Erasmus University, Netherlands; Hume Field of the University of Queensland, Australia; Sai Kit Lam of the University of Malaya, Malaysia; Carlos Das Neves of the Norwegian Veterinary Institute, Norway; Malik Peiris of the University of Hong Kong, China; Stanley Perlman of the University of Iowa; and Supaporn Wacharapluesadee of King Chulalongkorn Memorial Hospital and Chulalongkorn University, Thailand.

Experiments in cell cultures and mice showed that blocking the function of a specific enzyme present in all cells triggers a powerful innate immune response, the body’s first line of defense against any foreign invader. When challenged by several types of viruses in the study, this response dramatically lowered replication of viral particles and protected mouse lungs from damage. 

There are still several avenues to explore, but the scientists say the finding could help change the approach to developing antiviral medications. 

“Typically, in antiviral development, the saying is, ‘one bug, one drug,’” said Jianrong Li, co-senior author of the study and a professor of virology in The Ohio State University Department of Veterinary Biosciences and Infectious Diseases Institute. 

“A drug that can stimulate the immune system to have broad antiviral activities would be very attractive – one drug against multiple bugs would be an ideal situation.” 

The study is published in the journal Proceedings of the National Academy of Sciences.

This discovery was enabled in part by a technique the researchers used to map the precise location of an RNA modification they were studying, and to see which enzyme made the modification. The mapping led them to determine that this enzyme’s work happens not in viruses, but in mammal hosts that viruses want to infect. 

“If you can detect the modification, then you can study it and target it. But it took a while to figure this out – in the beginning of the pandemic, a lot of people, including our lab, were studying RNA modifications in hosts and viruses,” said co-senior author Chuan He, John T. Wilson Distinguished Service Professor of chemistry, biochemistry and molecular biology at the University of Chicago. “It turns out the key here is not a viral RNA modification, but a host RNA modification, and it triggers a host immune response.” 

Viruses tested against the immune response in this study included two that can cause severe respiratory infections in infants and the elderly, human respiratory syncytial virus and human metapneumovirus, as well as a mouse respiratory virus called Sendai virus, the vesicular stomatitis virus found in cattle and the herpes simplex virus, a DNA virus. Replication and gene expression of all of these viruses were significantly reduced when the enzyme was blocked, and the researchers said preliminary data from earlier studies in cell cultures suggested the SARS-CoV-2 virus could be similarly controlled by this antiviral strategy. 

The RNA modification itself, known as cytosine-5 methylation, or m⁵C, is actually what needs to be altered to trigger the immune system response. It is one of roughly 170 known chemical modifications on RNA molecules in living organisms that affect biological processes in a variety of ways. 

In lieu of targeting the modification, researchers were able to inhibit the function of a key enzyme in that process, called NSUN2, to stop the RNA change. Suppressing NSUN2 using gene knockdown techniques and experimental agents, they found, sets off a cascade of cell activities that leads to robust production of type 1 interferon, one of the most potent fighters in the innate antiviral response. 

“Amazingly, blockage of NSUN2 almost completely shuts down the replication of vesicular stomatitis virus, a model virus that normally kills the host cells within 24 hours and replicates to a very high titer, and strongly inhibits both RNA and DNA viruses,” said study co-first author Yuexiu Zhang, a PhD student in Li’s lab. 

It turns out that blocking NSUN2’s function in cells exposes RNA snippets that, despite belonging to the host, are seen as foreign invaders, which triggers the type 1 interferon production. Once available at this high level, the protein will stop the real threat: viruses trying to cause infection.

The researchers verified this sequence of events during experiments in multiple types of cells and human lung models before observing the effects of blocking NSUN2 in mice. 

“We compared NSUN2-deficient mice with wild-type mice to see how the viruses act,” Li said. “Once we inhibited NSUN2, viral replication in the lung decreased and there was less pathology in the lung, and that correlated with enhanced type 1 interferon production. 

“This finding in mice and our other experiments proved that NSUN2 is a druggable target.” 

Next steps include developing a drug designed specifically to suppress NSUN2’s function, the researchers said. 

This study was supported by grants from the National Institutes of Health and the Howard Hughes Medical Institute, where He is an investigator. 

Li-Sheng Zhang, a postdoctoral researcher in He’s lab, was co-first author of the work. Additional co-authors include Mijia Lu, Elizabeth Kairis, Valarmathy Murugaiah, Jiayu Xu, Rajni Kant Shukla, Xueya Liang, Estelle Cormet-Boyaka and Amit Sharma of Ohio State; Qing Dai and Zhongyu Zou of the University of Chicago; Phylip Chen and Mark Peeples of Nationwide Children’s Hospital; and Jianming Qiu of the University of Kansas Medical Center.

He is a scientific founder of the drug-development company Accent Therapeutics, and Li and He have filed a provisional patent.

The findings suggest a surveillance program is warranted for citizens in Africa who may be at risk for exposure to the virus. But the study teaches a much larger lesson as well, researchers say: It’s never too early to start preparing for the next animal virus to come along and unexpectedly cause disease in people.

“There are a lot of unknown animal viruses out there that may pose risk to humans,” said Cody Warren, first author of the study and assistant professor of veterinary biosciences at The Ohio State University. 

“We need to be prospectively looking at animal viruses that have been ignored to see if they have the capacity to replicate in human cells. If they do, will we continue to ignore them? I don’t think we should,” he said. 

Warren completed this work at the University of Colorado Boulder as a postdoctoral researcher in the lab of senior author Sara Sawyer, professor of molecular, cellular & developmental biology. 

The study is published online today (Sept. 30, 2022) in the journal Cell.

The virus the team studied is called simian hemorrhagic fever virus (SHFV), which exists in its natural reservoir, wild African monkeys, without causing any observable signs of disease. It is associated with several deadly outbreaks in rhesus monkey colonies worldwide, showing that the virus can kill when it enters a non-natural host. No known human infections have been documented.

SHFV is one of a dozen or so simian arteriviruses that have been identified to date circulating in wild African monkeys. Other labs’ previous research on arterivirus strains that infect pigs gave Warren and colleagues a good starting point for their investigation into what kind of protein SHFV uses to gain access to host cells.

In this new study, researchers determined in cell culture experiments that a receptor protein known as CD163, located inside a compartment within a cell’s cytoplasm, acts as a gatekeeper for this virus to infect a cell. And it turns out that human immune cells called myeloid cells circulating in the blood and our bodies’ tissues contain this very same receptor. But the human similarities to the primate hosts for this virus don’t stop there. 

“We took another leap forward to show that human genetics were fully compatible with replicating and producing this virus,” Warren said. “Just because a virus can replicate in a monkey cell doesn’t mean it can in human cells. But in this case, it does.” 

Preliminary findings in multiple cell lines also suggested that while SHFV is using humans’ cell machinery to make copies of itself and spread, the virus is simultaneously subverting the host cells’ ability to fight back – scientists showed that SHFV could readily grow in cells that are responsive to interferon, a cell’s early defense mechanism against viruses.

“We don’t yet understand how this virus interacts with the whole system – can it take down the entire interferon defense system, or simply evade it?” Warren said. “Those are key questions we have moving forward.” 

Warren’s lab focuses on animal viruses that have potential to spill over to humans, and on understanding the kinds of hurdles these viruses must overcome in order to infect a new host species. Staying ahead of viruses in this way ideally would position scientists to respond with surveillance testing capabilities and therapies when such a spillover event occurs. 

The history of HIV looms large in Warren’s mind: Simian immunodeficiency virus (SIV) first crossed from African monkeys into apes, a step that ultimately led to the evolution and emergence of the pandemic strain of human immunodeficiency virus that causes AIDS. 

“We didn’t know SIV existed prior to the emergence of HIV – it was a complete unknown,” Warren said. “Since HIV, there has been an onslaught of virus after virus jumping from animals to infect people: SARS, MERS, Zika, West Nile, SARS CoV-2. This is likely something that is going to continue. 

“If SHFV were to cause an outbreak tomorrow, we wouldn’t have therapeutics, but we now can prospectively look at potential drugs that could stop these viruses,” he said. “Just because animal viruses are obscure doesn’t mean they should remain in obscurity. Things can change very quickly, as HIV has taught us.” 

This work was supported by the National Institutes of Health, the Burroughs Wellcome Fund and the Laulima Government Solutions/National Institute of Allergy and Infectious Diseases (NIAID). Jens Kuhn of NIAID was a co-corresponding author of the study.

Data from two adults in the same study who had a dramatic loss in antibodies but received a second booster showed that the second dose completely restored antibodies to protective levels.

“The message from our study is clear: If you have a dramatic loss of antibodies from the first booster, you definitely need a second booster to get antibodies back,” said Shan-Lu Liu, senior author the study and a virology professor in the Department of Veterinary Biosciences at The Ohio State University.

“It’s not surprising to us to find that we see this antibody decline – in general, vaccine-induced antibodies decline over time regardless of the virus, and especially if the virus changes over time in the way that SARS-CoV-2 has changed,” said Liu, also a professor in the Department of Microbial Infection and Immunity.

The study is published as a letter to the editor today (Sept. 7, 2022) in the New England Journal of Medicine. 

Researchers tested serum from 46 health care professionals who had received the initial two-dose mRNA vaccine course and, later, one booster shot between one and nine months before their antibody levels were tested against the parent SARS-CoV-2 virus and the most dominant omicron variants, including BA.5, among recent and current infections. The team also had access to samples from two vaccinated health care workers who had received the second booster shot.   

Using statistical modeling to account for the variable passage of time after the first boosters among the sample population, researchers estimated an average rate of decline in neutralizing antibody levels of nearly 18% against the parent virus, 19.5% against the BA.1 omicron variant, 18.4% against BA.2.12.1 and 19.6% against BA.4/5 every 30 days. Put another way: Neutralizing antibody titers were about 1.7 times higher one to three months after the first booster than the levels detected seven to nine months after the shot, dropping about 50% within 95-108 days. 

The declines were slightly less steep in vaccinated health care workers who had also had a COVID-19 infection, but the differences were not significant by statistical standards, said co-first author John Evans, a PhD candidate in Ohio State’s Molecular, Cellular and Developmental Biology Program who works in Liu’s lab. 

In fact, the data reinforced the need for booster shots even after a COVID-19 infection: “People with infection got a small antibody benefit from having had the infection,” Evans said. “But overall, there was not much of a difference in the rate of decline across variants.” 

“We had two people who had an almost complete loss of neutralizing antibodies against the current predominant BA.5 strain at three to four months following the first booster vaccination but showed a complete recovery of antibody response in the blood after the second booster,” said Panke Qu, the leading co-first author of the study and a PhD candidate in the College of Veterinary Medicine’s Comparative Biomedical Sciences Graduate Program.

Previous work led by this research group showed the importance of a single booster to maintain protection against SARS-CoV-2, and omicron in particular. In a June 30 letter to the New England Journal of Medicine, Liu’s lab reported that the BA.4/5 and BA.2.12.1 omicron subvariants showed substantial resistance to immunity generated by the two-vaccine course compared to the earlier BA.1 and BA.2 subvariants, but that a single booster shot provided sufficient neutralizing antibodies against the two later subvariants three to four weeks after the vaccination. The team also reported in May that a COVID-19 booster shot provided strong and broad antibody protection against the range of omicron sublineage variants that were in circulation at the time.

“Now we’re showing how long the booster can last, which is a very important topic because people have been talking about whether and when to get a second booster,” said Liu, also associate director of Ohio State’s Center for Retrovirus Research and a program co-director of the Viruses and Emerging Pathogens Program in Ohio State’s Infectious Diseases Institute. 

The findings on reduced antibody durability from a single booster come a week after the Food and Drug Administration announced approval of an updated booster containing components from the original SARS-CoV-2 strain and the BA.4/5 omicron subvariant. 

“Our data tells us the second booster is really important, but of course we could not say anything about the new formulation,” Liu said. “We’re continuing our studies to analyze the neutralizing antibody response and durability of a second booster, including the new formulation.”

The researchers conducted the cell-culture studies using pseudoviruses – a non-infectious viral core decorated with different SARS-CoV-2 spike proteins on the surface structured to match known mutations. The method used to detect neutralizing antibodies in the blood samples accounted for the varying concentrations of antibodies produced by individuals. 

Statistical modeling was performed by co-author Lianbo Yu, a biostatistician in biomedical informatics and the Center for Biostatistics in Ohio State’s College of Medicine. Qu conducted most of the experiments in the current study. Julia Faraone, a PhD candidate of the Liu lab in Ohio State’s MCDB graduate program, was co-first author of the paper. Other co-authors, all from Ohio State, include Yi-Min Zheng, Qin Ma, Claire Carlin, Gerard Lozanski, Linda Saif, Eugene Oltz and Richard Gumina. 

This work was supported by anonymous donor funds and grants from the National Cancer Institute, a Glenn Barber Fellowship from Ohio State’s College of Veterinary Medicine, the Robert J. Anthony Fund for Cardiovascular Research, the JB Cardiovascular Research Fund and the National Institutes of Health.

Based on these combined findings in rodent experiments, Ohio State University scientists envision one day incorporating a coronavirus antigen into the measles-mumps-rubella (MMR) vaccine as a way to produce COVID-19 immunity in kids.

“We were pushing to make a vaccine for infants and children with the idea that if we could incorporate the mumps COVID vaccine into the MMR vaccine, you’d have protection against four pathogens – measles, mumps, rubella and SARS-CoV-2 – in a single immunization program,” said Jianrong Li, senior author of the study and a professor of virology in Ohio State’s Department of Veterinary Biosciences and Infectious Diseases Institute.

“If infants and children could develop immunity against COVID infection with the MMR vaccine, that would be great – no extra immunization needed.” 

The research is published today (July 27, 2022) in Proceedings of the National Academy of Sciences. 

To create the antigen that stimulates immunity in this vaccine candidate, researchers used a prefusion version of the SARS-CoV-2 spike protein – the shape it is in on the surface of the virus before the virus infects a cell. The spike was locked into this form by changing six of its amino acids to prolines, an inflexible amino acid.

Experiments comparing the six-proline protein, 6P, to a spike protein with two prolines, 2P, showed that the 6P induces significantly more neutralizing antibodies in mice and golden Syrian hamsters than the 2P version. These are the first published results showing the 6P antigen is more effective than the 2P spike protein with which the existing mRNA and adenovirus-based COVID-19 vaccines were created, the researchers said.

“The 6P antigen is about 8 1/2 times better than 2P. That’s a big deal – that’s a lot more antibody production, which may become important since it looks like the virus is going to keep on evolving,” said study co-author Mark Peeples, professor of pediatrics at Ohio State and a researcher at Nationwide Children’s Hospital in Columbus. 

Li, in collaboration with Peeples and Veterinary Biosciences Professor Stefan Niewiesk, previously led development of a measles-based COVID-19 vaccine candidate, but found that the mumps virus genome is even more amenable than measles to insertion of the spike protein gene. The study used a mumps vaccine strain that has been a component of the current MMR vaccine used in children since the 1960s. Once the 6P spike protein gene is inserted into the genome, the recombinant mumps virus replicates more slowly but is genetically stable and grows well in a World Health Organization-approved cell line for vaccine production. 

In animals, the live attenuated mumps vaccine modified in this way generated high levels of the 6P protein antigen, which triggered a strong immune response. 

Researchers compared the 6P spike protein vaccine candidate’s effectiveness to a 2P version in mumps, and to mumps without the spike protein, in mice engineered to be highly susceptible to mumps infection and in golden Syrian hamsters, a standard animal model for SARS-CoV-2 studies. In all experiments, the 6P vaccine produced the best results: higher concentrations of neutralizing antibodies after vaccination and, after introduction of a SARS-CoV-2 virus challenge a few weeks later, protection from lung damage and significantly fewer viral particles in the lungs and nasal cavity.

“We didn’t test the omicron variant for this paper, but the 6P vaccine induced neutralizing antibodies and T cell activity against several variants of concern and offered complete protection from disease caused by the parent SARS-CoV-2 virus and the delta variant,” Li said. “That’s very important and suggests that if we use the 6P spike protein for vaccine protection in the future, we can enhance a vaccine’s ability to protect humans.” 

In addition, the Li laboratory developed a rapid recombination system that enables the insertion of any antigen into a mumps or measles virus in a week. “With this technique, the MMR vaccine can be rapidly updated to protect against new SARS-CoV-2 variants as they emerge,” said study first author Yuexiu Zhang, a major contributor to this technology in Li’s lab. 

Researchers vaccinated study animals by both injections and by drops into the nose, an attractive needle-free option that, the results suggested, provides superior protection against the coronavirus by inducing not only system-wide antibodies, but a broad response of specialized antibodies called IgA on mucosal surfaces of the airways as well, Li said. 

“Like most of the injected vaccines, current mRNA and adenovirus-based COVID-19 vaccines primarily induce antibodies in the bloodstream. Unfortunately, antibodies in the bloodstream do not protect the mucosal site of the airways very well, which is the initial site of infection with SARS-CoV-2. In contrast to injected vaccines, our new intranasal vaccine may be the next generation of COVID-19 vaccine because it can induce robust IgA, which directly neutralizes the SARS-CoV-2 in the nose and airways tissues,” said co-author Prosper Boyaka, professor of mucosal immunology and chair of veterinary biosciences at Ohio State.

Added Peeples, “Having those antibodies on the surface of the airways could be a real advantage compared to hoping that some of the antibodies in the bloodstream will end up in the respiratory tract. 

The study findings also suggest that existing immunity to mumps from vaccination or previous infection may slow the initial stimulation of antibodies by the mumps-based COVID-19 vaccine, but does not prevent a strong, protective antibody response. 

“That finding suggests this candidate can be used not only in infants, but also in adults who have mumps antibodies,” Li said. 

The team began work on this mumps-based vaccine almost a year ago, long before the June 17 federal approval of COVID-19 vaccines for children 6 months or older. Given the MMR’s 50-plus-year history as a safe childhood vaccine, the scientists are now evaluating ways to modify the MMR with coronavirus antigens from multiple variants.

This study was supported by startup fund and bridge funding from Ohio State’s Department of Veterinary Biosciences and College of Veterinary Medicine, a seed grant from Nationwide Children’s Hospital and grants from the National Institutes of Health. Jason McLellan of the University of Texas at Austin provided the stabilized spike genes for this study.

Additional co-authors include Mijia Lu, Eunsoo Kim, Mohamed Shamseldin, Michelle Chamblee, Jesse Hall, Adam Kenny, Xueya Liang, Jacob Yount and Purnima Dubey of Ohio State; Mahesh K C, Piyush Dravid, Sheetal Trivedi, Supranee Chaiwatpongsakorn, Satyapramod Srinivasa Murthy, Himanshu Sharma and Amit Kapoor of Nationwide Children’s Hospital; and Chengjin Ye, Jun-Gyu Park and Luis Martinez-Sobrido of the Texas Biomedical Research Institute.

The Ohio State Innovation Foundation and the Abigail Wexner Research Institute at Nationwide Children’s Hospital have filed a provisional patent application for the mumps virus-based SARS-CoV-2 vaccine.

The study focused on protection against a recently identified variant of the Hendra virus, which, along with Nipah virus, has been responsible for deadly animal and human infection outbreaks in the Eastern Hemisphere. The 2011 movie Contagion depicts a fictional viral outbreak traced to an infected pig that is modeled on the Nipah virus.

The Hendra variant, identified in two fatally diseased horses and sick bats in Australia, featured dramatic genetic changes from the original virus – which created a sense of urgency among scientists to learn how existing countermeasures stack up against the restructured pathogen.

Researchers screened and determined in cell studies that several previously developed monoclonal antibodies designed to neutralize the original virus are also effective against the variant. The team also designed an additional antibody that could join three others in a powerful cocktail that would leave the virus with minimal ability to further mutate its way out of antibody recognition. 

“These four antibodies can bind simultaneously, which is important for preventing future escaping mutants,” said co-lead study author Kai Xu, assistant professor of veterinary biosciences at The Ohio State University.

“If you have only one or two antibodies, the virus can easily develop a mechanism to escape antibody recognition. If you have more antibodies in a cocktail developed as a therapeutic, it will decrease the chances of an escape mutant by many orders of magnitude. 

The study was published online recently in Proceedings of the National Academy of Sciences. 

Both Hendra and Nipah viruses can cause fatal disease in humans, horses, pigs and other mammals, and are transmitted between humans and animals. The flying fox, a bat species, is considered the viruses’ natural host. The very similar pathogens, discovered in the 1990s in Australia and Malaysia, respectively, cause severe respiratory symptoms and brain inflammation that lead to death in up to 95% of those infected. 

“Initially people thought these viruses might not mutate so much – their genome is largely stable, so it appeared that a countermeasure like an antibody, drug or vaccine could totally prevent them. But that’s not the case – just like SARS-CoV-2, a vaccine alone can’t win the war. The virus constantly evolves to adapt to a new host,” Xu said.

In a series of experiments conducted in a virus system lacking the pathogenic gene, the researchers first found that the variant, known as HeV-g2, attaches to the same receptor as the original HeV virus to enter host cells, and with the same strength. The variant, like the original, uses two proteins to get in. 

A total of six monoclonal antibodies – three for each entry protein – that were previously developed to attach to matching “footprints” on both Hendra and Nipah viral surface proteins were found to neutralize the HeV-g2 variant nearly as well as they blocked the original viruses. In earlier studies, post-infection treatment with these antibodies protected numerous animal species against lethal doses of Hendra and Nipah viruses. 

To provide even further protection, the researchers developed an additional antibody to be combined with three others that neutralize one of the two viral proteins that gain access to host cells.

“We know after precise atomic modeling and binding studies that these four antibodies, the new one plus the three developed previously, are compatible to each other and can bind at the same time,” Xu said. “You don’t want them to compete or interfere with each other – and you want that kind of combination as a cocktail for therapeutic development.”

A resulting monoclonal antibody treatment would be used after exposure to the virus. The researchers also tested the effectiveness of an existing Hendra virus vaccine candidate in two rhesus macaques, and found in blood drawn 28 days after the last of three injections that the vaccine generated a neutralizing antibody response in the animals against the HeV-g2 variant. 

“These findings are proof of principle that antibodies are effective against the new variant and we can combine multiple antibodies for multivalent drug development,” Xu said. “And most important, we found that although the mutation is significant, the existing countermeasures are still effective.” 

Xu co-led the research with Christopher Broder of Uniformed Services University and David Veesler of the University of Washington. Yan Xu of Ohio State was a co-first author on the study. Additional co-author institutions include the Henry M. Jackson Foundation for the Advancement of Military Medicine; the U.S. Public Health Services Commissioned Corps; and the University of Sydney, Commonwealth Scientific and Industrial Research Organization, Equine Veterinary and One Health Epidemiology, and a private veterinary practice, all in Australia. 

This work was supported by the Ohio State University Comprehensive Cancer Center, Path to K Grant through the Ohio State University Center for Clinical & Translational Science, the National Institute of Allergy and Infectious Diseases, a Pew Biomedical Scholars Award, the Burroughs Wellcome Fund, the University of Washington, the National Institutes of Health, and the Australian Government Department of Agriculture, Water and the Environment.

Researchers tested neutralizing antibody levels against the BA.2 and BA.3 omicron variants and deltacron, a recombinant variant created by the exchange of genetic material between delta and omicron.

Results showed that a third mRNA vaccine dose was required to generate a high enough concentration of antibodies to neutralize BA.2 and deltacron, as well as other sublineage omicron variants, including the original, known as BA.1, and BA.1.1. Antibodies produced by just the two-dose series of mRNA vaccines were enough to neutralize BA.3 – a sign this variant is not likely to produce a new surge of omicron infections, researchers say.

These findings were published today (May 18, 2022) in a letter to the editor of the New England Journal of Medicine (NEJM), and in an article published in Cell Host & Microbe on April 25, by a group of Ohio State University researchers.

“Three doses is better for everything,” said Shan-Lu Liu, the senior author of both studies and a virology professor in the Department of Veterinary Biosciences at Ohio State.

Recent national data have suggested that the BA.2 variant constitutes about 90% of COVID-19 cases in the United States.

“People have been asking about the recombinant deltacron and also BA.3, and now we have an answer. And it’s good news,” said Liu, also associate director of the university’s Center for Retrovirus Research and a program co-director of the Viruses and Emerging Pathogens Program in Ohio State’s Infectious Diseases Institute.

“Based on the neutralization pattern, one booster shot can protect against BA.2 and deltacron, and we do not suspect that BA.3 will become predominant because it’s sensitive to neutralization by even two doses.”

In the study published in NEJM, researchers tested antibody levels in serum from 10 health care professionals at Ohio State’s Wexner Medical Center. After two vaccine doses, on average, antibody levels were 3.3-fold and 44.7-fold lower against BA.3 and deltacron, respectively, than those that neutralized the parent SARS-CoV-2 virus. After the booster, antibody levels were much higher against all variants tested, and the same antibody level reductions were 2.9-fold and 13.3-fold – showing a dramatic improvement in protection, particularly against deltacron.

Next examining blood samples from 18 patients in the ICU during the delta wave of the pandemic, the researchers found comparable levels of antibodies against the parent virus and BA.3, but 137.8-fold lower concentrations against deltacron compared to the parent virus. Blood samples from 31 hospitalized non-ICU patients during the omicron surge showed much better antibody protection against deltacron.

“The deltacron spike protein is structured similarly to other omicron variants, so people infected with omicron seem to have pretty good protection against deltacron. But people infected with delta had pretty weak protection against deltacron,” said John Evans, a PhD candidate in Ohio State’s Molecular, Cellular and Developmental Biology Program who works in Liu’s lab, and the first author of the new studies. “However, compared to the booster, infection by omicron provides much less protection against deltacron.”

The Cell Host & Microbe study tested antibody levels in serum from 48 health care professionals after their second mRNA vaccine dose and from 19 after their third mRNA vaccine doses, as well as 31 COVID-19 patients hospitalized during the omicron surge.

“For two vaccine doses, health care workers had very weak neutralizing antibody concentrations against the omicron variants, but after the booster it evened out,” Evans said. “The neutralizing antibody levels were still lower against omicron compared to the parental virus, but much more comparable across variants and much higher. So the booster provides much stronger protection and much broader protection.”

Patients with delta or omicron infection were similarly protected against their respective infectious variants, and a vaccine dose on top of infection improved protection. Among all of the health care professionals and infected patients studied, people who had received three vaccine doses had the broadest and strongest antibody protection overall.

The researchers conducted the cell-culture studies using pseudoviruses – a non-infectious viral core decorated with different SARS-CoV-2 spike proteins on the surface structured to match known mutations. The method used to detect neutralizing antibodies in the blood samples accounted for the varying concentrations of antibodies produced by individuals.

Liu and colleagues have completed additional tests of neutralizing antibody responses to the BA.2.12.1, BA.4 and BA.5 omicron sublineages, which are growing in dominance among newly reported cases. In a paper posted May 17 on the bioRxiv preprint server (and published in the New England Journal of Medicine on June 15), they described similar findings: A booster provided sufficient antibody concentrations against these three additional omicron subvariants, though to a lower extent than protection seen for BA.1 and BA.2, and previous omicron infection did not offer effective antibody protection against these variants. 

Liu’s lab is continuing this work, investigating how long antibody protection from the third booster shot lasts. Waning antibody levels seen in blood samples collected the furthest out from the third dose suggest there is a limit to the first booster’s durability, Liu said. 

“The virus is evolving, which is typical for all RNA viruses, and what we have seen is not surprising at all,” Liu said. “Especially when we have lots of variants in the population, anything can happen, including recombination between them. That’s why we still need to be very cautious. The best preparation is to get the second booster.”

This work was supported by anonymous donor funds, grants from the National Institutes of Health, the National Center for Advancing Translational Sciences, the Glenn Barber Fellowship from Ohio State’s College of Veterinary Medicine, the National Cancer Institute and the Robert J. Anthony Fund for Cardiovascular Research. Liu also acknowledged the generosity of the health professionals and patients who participated in the studies.

Additional co-authors on both papers include Cong Zeng, Panke Qu and Yi-Min Zheng of the Center for Retrovirus Research; Linda Saif of Ohio State’s Center for Food Animal Health; and Claire Carlin, Joseph Bednash, Gerard Lozanski, Rama Mallampalli, Eugene Oltz, Peter Mohler and Richard Gumina of Ohio State’s College of Medicine. Virus researchers Julia Faraone, Xue Zou and Kai Xu of Ohio State and Tongqing Zhou of the NIH were co-authors on the Cell Host & Microbe paper and the preprint.

The same Ohio State University lab found in a previous study, posted on the preprint server bioRxiv, that a third COVID-19 mRNA vaccine booster shot did produce effective levels of neutralizing antibodies against omicron. This study has not yet been peer-reviewed.

“Our new work shows that two doses of mRNA vaccine do not offer protection against omicron, and even having a breakthrough infection on top of vaccine does not help much. But our earlier study showed that the booster can really rescue the shortcomings of the two doses,” said Shan-Lu Liu, the senior author of both studies and a virology professor in the Department of Veterinary Biosciences at Ohio State.

The new research is published online as a First Release paper in the journal Science Translational Medicine.

The researchers examined antibodies in serum samples from 48 health care professionals with experimental versions of the parent virus and the alpha, beta, delta and omicron variants. Serum samples were collected pre-vaccination, three to four weeks after a first vaccine dose, three to four weeks after a second vaccine dose and six months after the second vaccine.

“There was a substantial increase in neutralizing antibodies after the second dose against every variant except the omicron variant,” said first study author John Evans, a PhD student in Ohio State’s Molecular, Cellular and Developmental Biology Program who works in Liu’s lab. “From the second dose to six months later, there was an at least five-fold drop in immunity, even against the parent virus.”

Neutralizing antibodies that block viral particles’ entry into host cells are considered the gold standard of protection against COVID-19 infection.

Twelve of the samples came from people suspected to have had a COVID-19 infection – at time points ranging from before vaccination to after two vaccine doses – based on a different kind of antibody testing. And though the findings suggested a breakthrough COVID-19 infection on top of vaccination increased immunity against most versions of the virus, antibodies from only one individual with previous infection reached levels that could put up a reasonable fight against omicron.

“Overall, nobody in this study had good immunity against omicron,” said Liu, also an investigator in the university’s Center for Retrovirus Research and a program co-director of the Viruses and Emerging Pathogens Program in Ohio State’s Infectious Diseases Institute.

The experimental viruses were what are called pseudoviruses – a non-infectious viral core decorated with different SARS-CoV-2 spike proteins on the surface structured to match known mutations in the variants studied.

The researchers used a special method to detect neutralizing antibodies in the health professionals’ blood samples to account for the varying levels of antibodies produced by individuals.

“Individuals did respond very differently to the first dose, and the same was true for the second dose,” Liu said.

Results also showed that people who received the Pfizer-BioNTech mRNA vaccine produced about two-fold lower levels of neutralizing antibodies than those who received the Moderna vaccine. Men also had significantly higher antibody levels compared to women against all variants over the post-vaccination time points.

Liu said the dramatic reduction in immunity six months after two vaccine doses and the earlier paper’s finding that a booster protects against omicron highlight how important a third shot is to avoiding infection.

“After the second vaccine dose, the neutralizing antibodies effective against omicron dropped 23-fold, but with a booster shot, immunity dropped only three- to four-fold – which is comparable to booster effectiveness previously reported against the delta variant,” he said. “Similar observations have been made by other labs.”

Additional co-authors of the Science Translational Medicine study, all from Ohio State, include Cong Zeng, Claire Carlin, Gerard Lozanski, Linda Saif, Eugene Oltz and Richard Gumina. Additional co-authors on the bioRxiv preprint include Panke Qu, Julia Faraone, Yi-Min Zheng, Joseph S. Bednash, Rama Mallampalli, Peter Mohler and Kai Xu of Ohio State and Tongqing Zhou from NIH.

This work was supported by anonymous donor funds, grants from the National Institutes of Health, the Glenn Barber Fellowship from Ohio State’s College of Veterinary Medicine, the National Cancer Institute and the Robert J. Anthony Fund for Cardiovascular Research.

Previous research led by the U.S. Department of Agriculture had shown evidence of antibodies in wild deer. This study, published today in Nature, details the first report of active COVID-19 infection in white-tailed deer supported by the growth of viral isolates in the lab, indicating researchers had recovered viable samples of the SARS-CoV-2 virus and not only its genetic traces.

Based on genomic sequencing of the samples collected between January and March 2021, researchers determined that variants infecting wild deer matched strains of the SARS-CoV-2 virus that had been prevalent in Ohio COVID-19 patients at the time. Sample collection occurred before the Delta variant was widespread, and that variant was not detected in these deer. The team is testing more samples to check for new variants as well as older variants, whose continued presence would suggest the virus can set up shop and survive in this species.

The fact that wild deer can become infected “leads toward the idea that we might actually have established a new maintenance host outside humans,” said Andrew Bowman, associate professor of veterinary preventive medicine at The Ohio State University and senior author of the paper.

“Based on evidence from other studies, we knew they were being exposed in the wild and that in the lab we could infect them and the virus could transmit from deer to deer. Here, we’re saying that in the wild, they are infected,” Bowman said. “And if they can maintain it, we have a new potential source of SARS-CoV-2 coming in to humans. That would mean that beyond tracking what’s in people, we’ll need to know what’s in the deer, too.

“It could complicate future mitigation and control plans for COVID-19.”

A lot of unknowns remain: how the deer got infected, whether they can infect humans and other species, how the virus behaves in the animals’ body, and whether it’s a transient or long-term infection.

The research team took nasal swabs from 360 white-tailed deer in nine northeast Ohio locations. Using PCR testing methods, the scientists detected genetic material from at least three different strains of the virus in 129 (35.8%) of the deer sampled.

The analysis showed that B.1.2 viruses dominant in Ohio in the early months of 2021 spilled over multiple times into deer populations in different locations.

“The working theory based on our sequences is that humans are giving it to deer, and apparently we gave it to them several times,” Bowman said. “We have evidence of six different viral introductions into those deer populations. It’s not that a single population got it once and it spread.”

Each site was sampled between one and three times, adding up to a total of 18 sample collection dates. Based on the findings, researchers estimated the prevalence of infection varied from 13.5% to 70% across the nine sites, with the highest prevalence observed in four sites that were surrounded by more densely populated neighborhoods.

White-tailed deer functioning as a viral reservoir of SARS-CoV-2 would likely result in one of two outcomes, Bowman said. The virus could mutate in deer, potentially facilitating transmission of new strains to other species, including humans, or the virus could survive in deer unmutated while it simultaneously continues to evolve in humans, and at some point when humans don’t have immunity to the strains infecting deer, those variants could come spilling back to humans.

How transmission happened initially in these deer, and how it could happen across species, are among the pending questions related to these findings. The research team speculated that white-tailed deer were infected through an environmental pathway – possibly by drinking contaminated water. Research has shown that the virus is shed in human stool and detectable in wastewater.

The white-tailed deer tested for this study were part of a population control initiative, so they are not a transmission threat.

Though there are an estimated 600,000 white-tailed deer in Ohio and 30 million in the United States, Bowman said this sampling focused on locations close to dense human populations and is not representative of all free-ranging deer.

This work was supported by the Ohio State Infectious Diseases Institute and the National Institute of Allergy and Infectious Diseases. In addition to USDA, NIAID, Ohio Wildlife Center and Cleveland Metroparks contributors, Ohio State co-authors include Vanessa Hale, Patricia Dennis, Dillon McBride, Jacqueline Nolting, Christopher Madden, Devra Huey, Margot Ehrlich, Jenessa Winston, Dubraska Diaz-Campos, Page Yaxley, Alexis McLaine, Risa Pesapane, Mark Flint, Jaylene Flint, Anastasia Vlasova, Scott Kenney, Qiuhong Wang, Linda Saif and Seth Faith.

Cell culture experiments showed that SARS-CoV-2, which causes COVID-19, limits the release of viral particles that can be inactivated by antibodies, instead staying tucked within cell walls and spreading between cells.

“It’s basically an underground form of transmission,” said lead author Shan-Lu Liu, a virology professor in the Department of Veterinary Biosciences at The Ohio State University and an investigator in the university’s Center for Retrovirus Research.

“SARS-CoV-2 can spread efficiently from cell to cell because there are essentially no blockers from the host immunity. Target cells become donor cells, and it just becomes a wave of spread, as the virus may not get out of the cells.”

Liu and colleagues found other revealing details about SARS-CoV-2: The spike protein on its surface alone enables cell-to-cell transmission, and yet the virus’s primary receptor on target cells – to which the spike binds – is not a necessary part of the cell-to-cell transmission operation. Additionally, they found that neutralizing antibodies are less effective against the virus when it spreads through cells.

The research is published today (Dec. 22, 2021) in the journal Proceedings of the National Academy of Sciences.

A major point of this study was comparing SARS-CoV-2 to the coronavirus behind the 2003 SARS outbreak, known as SARS-CoV. The findings help explain why while the first outbreak led to much higher fatality rates and lasted only eight months, we’re about to surpass the two-year mark of the current pandemic, with a majority of cases being asymptomatic, Liu said.

The comparison showed that the SARS-CoV that caused SARS in 2003 is more efficient than SARS-CoV-2 at what is called cell-free transmission, when freely floating viral particles infect target cells by binding to a receptor on their surface – but also remain vulnerable to antibodies produced by previous infection and vaccines. SARS-CoV-2, on the other hand, is more efficient at cell-to-cell transmission – which makes it harder to neutralize with antibodies.

The viruses’ differing efficiencies were first demonstrated in experiments using pseudoviruses – a non-infectious viral core decorated with both kinds of coronavirus spike proteins on the surface.

“The spike protein is necessary and sufficient for both SARS-CoV-2 and SARS-CoV cell-to-cell transmission because the only difference in these pseudoviruses were the spike proteins,” said Liu, also a program director of the Viruses and Emerging Pathogens Program in Ohio State’s Infectious Diseases Institute.

Looking more deeply into those differences, the researchers found that SARS-CoV-2 is also more capable than SARS-CoV at initiating fusion with a target cell membrane, another key step in the viral entry process. And that stronger fusion action was associated with the virus’s enhanced cell-to-cell transmission.

Paradoxically, too much cell membrane fusion leads to cell death and can actually interfere with cell-to-cell transmission, Liu also found.

The team then turned to the role of the ACE2 receptor, a protein on cell surfaces that acts as the gateway for entry of the virus that causes COVID-19. The researchers found, unexpectedly, that cells with no or low levels of ACE2 on their surfaces can be penetrated by the virus, enabling robust cell-to-cell transmission.

“There is no perfect correlation between SARS-CoV-2 infection and the level of ACE2,” Liu said. “ACE2 may be needed for initial infection, but once infection is established, the virus may not need ACE2 anymore because it can spread from cell to cell.”

Finally, in experiments testing blood samples from human COVID-19 patients against the authentic SARS-CoV-2 virus, researchers determined that the virus could evade an antibody response through cell-to-cell transmission, but that antibody neutralization of the virus in the cell-free transmission mode was effective.

“We were able to confirm cell-to-cell transmission is not sensitive to inhibition from antibodies from COVID patients or vaccinated individuals,” Liu said. “Cell-to-cell transmission’s resistance to antibody neutralization is probably something we should watch for as SARS-CoV-2 variants continue to emerge, including the most recent, Omicron. In this sense, developing effective antiviral drugs targeting other steps of viral infection is critical.”

There are still many unknowns, including the exact mechanism the virus uses to spread from cell to cell, how that may influence individuals’ responses to viral infection, and whether or not efficient cell-to-cell transmission contributes to the emergence and spread of new variants. Liu’s lab is planning additional studies using the authentic virus and human lung cells to further explore these questions.

This work was supported by grants from the National Institutes of Health and funds from an anonymous private donor to Ohio State.

Ohio State co-authors include Cong Zeng, Jack Evans, Tiffany King, Yi-Min Zheng, Eugene Oltz, Linda Saif and Mark Peeples, also a researcher at Nationwide Children’s Hospital. Sean Whelan of the Washington University School of Medicine also contributed.

GOHi and several Ohio State University partners will collaborate with the Government of Ethiopia to strengthen workforce capacity within a network of hospitals and a central laboratory as well as processes and systems with the goal to provide rapid detection, response and prevention of infectious disease threats.

As part of a five-year cooperative agreement through 2026, the grant will provide $800,000 in funding for the first year, and allows GOHi to continue its One Health-related work to bolster infection prevention and control measures, implement laboratory quality assurance to maintain high levels of accuracy and proficiency, and build capacity through training, mentoring and evaluation. Funding beyond the first year will be based on satisfactory programmatic progress and availability of funds.

GOHi’s project is part of the Global Action in Healthcare Network, one of two global networks established by the CDC to combat antimicrobial resistance and infectious diseases that will span more than 50 countries worldwide. The CDC has awarded $22 million to nearly 30 organizations around the globe.

The continued use of antimicrobials in health care facilities has resulted in the emergence of a variety of antimicrobial resistant pathogens over the past decade, which has caused the rapid spread of disease, serious health conditions and patients dying from preventable and treatable diseases.

“Antimicrobial resistance (AMR) is an increasing threat to global health security, having economic, social and political ramifications worldwide, and puts an excess burden on resource-poor countries, such as Ethiopia,” said Wondwossen Gebreyes, executive director of GOHi. “To combat AMR, it is critical to implement a collaborative One Health approach that brings together multiple disciplines working globally to address the spread of disease, promote health and emphasize the connection among humans, animals, plants and the environment.”

Since 2009, GOHi has been working extensively on a range of One Health-related projects targeting laboratory, surveillance and health care workforce capacity through applied education, training and research globally.

Over the next five years, the GOHi team and the Ethiopian government will work to ensure the continued collection and entry of quality surveillance and laboratory data to monitor trends and trigger alarms for potential outbreaks at facilities. Successful implementation of these and other activities will improve network preparedness and response to antimicrobial resistance threats.

This award is GOHi’s second from the CDC this year. In April, GOHi received $5.61 million in funding from the CDC to fight the COVID-19 pandemic and strengthen Ethiopia’s public health system capacity for small- and large-scale disease outbreaks and emergencies.

Ohio State partners on this new initiative include the colleges of Medicine (Shu-Hua Wang, Joan-Miquel Balada-Llasat and Xia Ning), Veterinary Medicine (Gebreyes and Zelalem Mekuria) and Public Health (Saira Nawaz and Jiyong Lee), as well as the Global One Health, LLC in Ethiopia (Getnet Yimer and Ebba Watola),

RSV, or respiratory syncytial virus, accounts for an estimated 14,000 deaths and 177,000 hospitalizations annually among U.S. adults over 65. RSV is problematic for older people in part because an initial infection isn’t effective at inducing a long-term immune response to the virus.

The study showed that geriatric cotton rats given ibuprofen for a week before initial infection with RSV cleared the virus more quickly than those that didn’t receive ibuprofen. When researchers re-infected those ibuprofen-treated rats a month later, the elderly animals were completely protected from the virus.

In contrast, young adult cotton rats were able to clear the virus fairly effectively and mount a suitable immune response to re-infection a month later – but got no benefit from the ibuprofen.

With ibuprofen showing effectiveness only in geriatric animals, the findings suggest that the drug improves the immune response by lowering aging-related inflammation. Though there is still a lot to learn, the Ohio State University researchers speculate that the reduced inflammation restored the function of specific immune cells needed to clear the virus.

“For a long time, people have thought that certain immune cells get burned out and can’t function properly any longer. And then we started treating against inflammation, and suddenly the old cells can do their job like young cells,” said senior study author Stefan Niewiesk, professor of veterinary biosciences at Ohio State.

“It only works in old animals, and in older people we see a lot of inflammation and related chronic disease. When we think about what the pathways are to fix RSV infection in the elderly, this study opens a door to the possibilities.”

The research appears in the November 2021 issue of the journal Virology.

Niewiesk and first author Olivia Harder, a veterinary biosciences research associate at Ohio State, cautioned that it’s far too soon to consider taking ibuprofen to prevent RSV infection. Long-term use of ibuprofen, a nonsteroidal anti-inflammatory drug (NSAID), can damage the kidneys and gastric lining and slows down blood clotting time.

In this study, the researchers compared the influence of ibuprofen pre-treatment on groups of sexually mature adult cotton rats, younger than 2 months old, and geriatric rats between 9 and 15 months old. Cotton rats are a good model for the study of RSV because the virus grows well in both the lungs and nose as it does in humans, but the rats don’t experience symptoms of illness.

The drug was added to their chow and drinking water for one week before they were first infected with RSV. The estimated ibuprofen dose was intended to resemble the 200 to 400 milligrams humans might take every four to six hours.

Though ibuprofen was linked to more rapid viral clearance in old rats, the scientists were particularly interested in the animals’ response to re-infection a month later.

“We wanted to see if they could mount an immune response,” Harder said. “In the geriatric cotton rats we had treated with ibuprofen, there was no growth of the virus with the second infection.”

In examinations of cells and tissue, Harder found that ibuprofen did not improve the antibody response in the old rats or kill the virus. The analysis identified a specific type of immune cell, called CD8+ (cytotoxic) T cells, that were critical to the geriatric rats’ ability to clear the virus because they remove infected cells. When the team used a technique to deplete CD8+ T cells in the old rats, it took the animals longer to clear the virus and any positive effects of the ibuprofen were lost – suggesting the ibuprofen had a restorative effect on those cells.

Previous research has suggested that other immune cells responsible for stimulating T cell production get sluggish in old animals, and that inflammation-related molecules are behind their slowed reactivity.

“If you take those inflammatory molecules away with ibuprofen, these immune cells would migrate normally and we’d have stimulated CD8+ T cells. That’s the idea, but we still haven’t confirmed the mechanism,” Niewiesk said. “We’re also asking what part of the inflammatory cascade does ibuprofen affect? How does inflammation look in these old animals – is it always there and then increased with infection? That’s where we’re looking at the moment.”

RSV vaccines are in development, but there is no treatment – with the exception of an expensive prophylactic antibody available to prevent severe symptoms only in at-risk infants.

Niewiesk hopes to eventually identify a more targeted agent for the aging population that could have a similar protective effect against RSV without any risks.

“This isn’t a case where someone should take ibuprofen daily for the whole winter. There are too many side effects,” he said. “But if there was something to take post-infection, after you feel sick – that would be great.”

This work was supported in part by the National Institute of Allergy and Infectious Diseases. Former graduate student Margaret Martinez also worked on the study.

Researchers traced this link in mice to gut bacteria that, after exposure to ingested cadmium, over-produced an enzyme that degrades vitamin D – effectively creating conditions that mimic vitamin D deficiency. In terms of clinical effects, the mice sensitized to a specific allergen that consumed cadmium produced high levels of antibodies against the allergen as well as immune cells that increased their respiratory symptoms.

Separate epidemiological research has shown an association in children between vitamin D deficiency and higher susceptibility to asthma and other allergy symptoms. And a Congressional report released on Sept. 29 disclosed an unexpected source of cadmium in kids, announcing that dangerous levels of toxic heavy metals, including cadmium, had been detected in several brands of baby food.

“The problem is, because cadmium doesn’t degrade easily – it has a half-life in the body of at least 15 years – if you are chronically exposed to low doses, it accumulates over time,” said Prosper Boyaka, professor and chair of veterinary biosciences at The Ohio State University and senior author of the study. “It’s also not something we can easily avoid being exposed to because it can remain in air, soil and water.”

Most people ingest the natural element cadmium, a heavy metal used for batteries and making pigments, by eating plant and animal foods that have absorbed the pollutant or drinking contaminated water. The Environmental Protection Agency lists cadmium among eight metals considered extremely toxic at small concentrations.

Boyaka and colleagues found that an experimental compound that inhibits the activated enzymes reduced the allergic response in mice that ingested cadmium.

“We proposed two possible strategies in the paper,” Boyaka said. “One is vitamin D supplementation, but that has to happen before cadmium exposure has caused a heightened allergic reaction, so the question would be when to use a supplement. We also propose targeting those enzymes as a way to prevent the heightened allergic response.”

The research was published recently in the journal Mucosal Immunology.

Environmental pollutants such as cadmium and lead are considered contributors – along with genetic predisposition and overuse of antibiotics – to the growing number of people with allergies, but the mechanisms behind that association aren’t well-understood, Boyaka said.

In this study, researchers added what is considered a “subtoxic” dose of cadmium to drinking water consumed by groups of mice for 28 days. Using a mouse model that simulates human genetic predisposition to an egg allergy, the team exposed the mice to an egg protein to test their allergic response.

Mice that drank water containing subtoxic doses of cadmium and then were exposed to the allergen had a stronger allergic reaction – in the form of internal inflammatory actions and allergy symptoms – than the response in control mice.

“Our hypothesis was that cadmium would change the microbe population in the gut because we know that dysbiosis, or a change in the microbiota, can drive allergic responses. And yes, giving those tiny, tiny doses of cadmium in the drinking water did change allergic sensitization,” Boyaka said.

The effect was known to occur in the gut, as expected, because germ-free mice, which lack intestinal microbes, that were orally exposed to cadmium did not over-produce the antibody that causes an allergic response. But the researchers determined the cadmium wasn’t killing cells or making the intestines leaky. A series of experiments linked cadmium in the gut to production of inflammatory molecules – as well as to stimulation of the two enzymes that degrade vitamin D.

“That’s the main finding – after exposure to subtoxic doses of heavy metals, the pollutants remain in soft tissue, including in the gut. And what they do is make cells more reactive. In the gut, specifically, bacteria will make certain cells produce more of the enzyme that degrades vitamin D,” Boyaka said. “That’s a connection that we did not know before.”

Boyaka’s lab is now investigating potential compounds that could be used to block the vitamin D-degrading enzymes.

This work was supported by grants from the National Institutes of Health and the National Center for Advancing Translational Sciences. The researchers conducted experiments using the Ohio State Center for Clinical and Translational Science Mass Spectrometry and Proteomics Core.

Co-authors, all from Ohio State, include Eunsoo Kim, Astrid Bonnegarde-Bernard, Stephen Opiyo, Marisa Joldrichsen, Zayed Attia, Brian Ahmer and Estelle Cormet-Boyaka.

The College of Medicine, College of Nursing, College of Optometry, College of Public Health and College of Veterinary Medicine were recognized. Ohio State is the only academic institution in the country to have five colleges honored with the award.

“The recognition bestowed upon these five colleges is a testament to our collective commitment to diversity and inclusion,” said Melissa Gilliam, executive vice president and provost at Ohio State. “The entire university community benefits from this recognition as we strive to live up to our land-grant university values.”

The Health Professions HEED Award process consists of a comprehensive and rigorous application that includes questions relating to the recruitment and retention of students and employees, leadership support for diversity, and other aspects of campus diversity and inclusion. Ohio State diversity and inclusion efforts include: 

College of Medicine: The College of Medicine is executing an anti-racism action plan that includes a review of medical curriculum to remove racism and bias. In addition, the college appointed vice chairs of diversity, equity and inclusion in all 26 departments and implemented best practices for inclusive faculty searches, which includes creating a standing search committee within each department whose members will complete implicit bias mitigation training.

College of Nursing: In May 2021, the Office of Diversity, Equity and Inclusion at the College of Nursing held virtual graduate entry welcome panels for incoming graduate entry students of all backgrounds. The panels were designed to enhance incoming graduate students’ understanding of psychosocial, academic, clinical, professional practice and research aspects of the nursing student experience.

College of Optometry: With an aim of cultivating interest in optometry as a career and creating a pipeline of students who exhibit inclusive thinking and behavior, the College of Optometry offers the I-DOC – Improving Diversity in Optometric Careers – summer program for underrepresented diverse students. During this program, students learn about health disparities in optometry, explore different careers in optometry and review how to be successful learners and applicants. 

College of Public Health: In 2020, the College of Public Health launched a trio of scholarships designed to increase diversity and equitable access to public health education: the Ruth Ella Moore, Dr. Amy Acton Future Leaders of Public Health and the Multicultural Public Health Hood scholarships. The scholarships will be awarded annually with the intention of championing those who are first-generation or trailblazing students, those who have overcome significant challenges and those students who seek to advance Black Americans. 

College of Veterinary Medicine: The College of Veterinary Medicine has a strategic focus on recruiting underrepresented minorities and first-generation students by reducing barriers to enter the profession, providing engaging learning opportunities and creating an academic community that fosters inclusion. This includes the creation of several new scholarships and the use of a virtual recruitment platform that provides an opportunity to connect with historically Black colleges and universities for virtual admissions sessions.

Ohio State will be featured in the December 2021 issue of INSIGHT into Diversity magazine. 

As the word of these discoveries spread in the scientific community, Gebreyes, a professor of veterinary preventive medicine, was invited to help researchers in Latin America and Eastern Africa, and later in Asia and Europe, with investigations of multidrug-resistant organisms and establishing molecular epidemiology capacity.

His body of molecular epidemiology research, and the broad worldwide partnerships that followed, were cited today as the National Academy of Medicine (NAM) announced Gebreyes’ election to the academy’s 2021 class of inductees, considered one of the highest honors in the fields of health and medicine.

Gebreyes was recognized by the academy for his leadership in molecular epidemiology and global health and for providing fundamental insight into how animal agricultural and environmental systems influence public health, community development and livelihood worldwide.

“I’m very humbled. But the work starts now – it’s a place for service and more action,” he said. “I’m looking forward to serving the collective scientific community and communities in general.”

The NAM was founded in 1970 as the Institute of Medicine and is one of three academies that make up the National Academies of Sciences, Engineering and Medicine in the United States. The academies are private, nonprofit institutions that work outside of government to provide objective advice on matters of science, technology and health.

The 2021 inductees represent the most diverse class of new members to date, composed of approximately 50% women and 50% racial and ethnic minorities, according to National Academy of Medicine President Victor J. Dzau. New members are elected by their peers in recognition of outstanding achievement and their contributions to health and society.

Also the executive director of Ohio State’s Global One Health initiative (GOHi), Gebreyes has been a driving force behind Ohio State’s global impact for years in the area of One Health, a strategy to more fully understand and address the links between animal, human and plant health and the environment.

A significant part of GOHi’s work involves research, training and outreach capacity-building in partner countries – an initiative that traces back to Gebreyes’ initial international collaborations on studies of antimicrobial resistance.

“We noticed that there was a severe lack of science and investigation and skilled personnel to do this kind of molecular work,” he said. “I started to teach researchers how to do molecular epidemiology investigations in many countries, and to build capacity on how to do molecular epidemiology work on drug resistance.”

By 2010, Gebreyes received the first National Institutes of Health Fogarty International Center research training grant focusing on antimicrobial resistance in Ethiopia, Kenya and Tanzania – a program that remains active today.

“We have trained 28 PhDs so far through the program in molecular epidemiology, and they are now the people in their countries doing diagnostics of COVID-19 because the same tools are used,” he said.

GOHi involves faculty and students in colleges and institutes across Ohio State and has added basic and applied research and community outreach to the training and capacity-building core mission. External partners and funders include the Centers for Disease Control and Prevention, the U.S. Agency for International Development, the Bill and Melinda Gates Foundation and dozens of agencies in collaborating countries. The initiative is widely recognized for its global leadership and effectiveness, and Gebreyes now serves in an advisory capacity to other U.S. universities working to establish similar consortia.

“I enjoy tackling grand challenges, and the need for broader global capacity-building remains a priority,” he said. “We have learned from COVID-19 that the world is severely underprepared to address an infectious disease pandemic.”

Gebreyes is also currently an American Council on Education fellow at Michigan State University where, in addition to apprenticing in higher education leadership, he is collaborating on a project aimed at ensuring the U.S. higher education system of the future proactively addresses societal challenges, especially those targeted by the United Nations Sustainable Development goals and that remain critical in Ohio and other states.

“Perhaps now more than ever, we understand how interconnected the world really is. With his scientific knowledge and extraordinary international partnerships, Dr. Gebreyes will no doubt be an important contributor to the National Academy of Medicine’s deliberations and policy recommendations,” said Grace Wang, executive vice president for research, innovation and knowledge at Ohio State. “We are so proud that he is being recognized for his work to improve global public health.”

Gebreyes’ election brings the total number of current Ohio State National Academies members to 33, including one professor who is a member of two academies. Gebreyes is one of 10 National Academy of Medicine members at Ohio State.

The key to their newly discovered power: enhancing the signals sent within immune cells to unleash one of the most potent fighters in the innate antiviral response, proteins called type I interferons.

Experiments in cell cultures showed that the cell-signaling activities of these proteins, from the SERINC family, helped protect cells from infection by HIV, Ebola and Zika viruses. The researchers are currently testing whether this function is also effective against SARS-CoV-2, the virus that causes COVID-19.

It’s one thing for a protein to act directly against a specific virus, as SERINC5 has been shown to do by incorporating itself into the HIV particle during viral production. But it’s another thing entirely to reveal that a protein has an enhancing effect on an essential antiviral cell signaling pathway upon infection of host cells, said senior study author Shan-Lu Liu, professor of virology in the Department of Veterinary Biosciences at The Ohio State University.

“Viruses can get around direct antiviral effects,” said Liu, also an investigator and associate director in Ohio State’s Center for Retrovirus Research. “But if this protein can also modulate key pathways without acting directly on the virus, then a virus may have limited capacity to counteract it.

“If this family of molecules can do this in animals and humans, then you may think about whether it could be used in a broad antiviral therapy.”

The research is published today (Sept. 14, 2021) in the journal Science Signaling.

Liu has studied what he calls the “evolutionary arms race” between HIV and the human immune system for years, and led a 2019 study that described how an HIV protein called Nef can get the upper hand in that fight – specifically by interfering with SERINC proteins in virus-producing cells.

In this study, Liu and colleagues took a closer look at SERINC5’s functions in target cells during the next viral infection stage. They found that in addition to boosting the signals that produce type I interferons, SERINC5 has the same effect on signals that generate another component of the early antiviral response – NF-kB inflammatory cytokines.

SERINC5 doesn’t modulate this signaling system on its own – it joins forces with other proteins called MAVS and TRAF6. But some of its exact behaviors are still a mystery, Liu said.

In cell cultures in which viral infection had been initiated, the researchers observed that SERINC5 relocates itself from the cell surface to just outside the membrane around the mitochondria, a cell’s power center – an unusual move. Once there, SERINC5 teamed up with MAVS and TRAF6 to form a big, multifaceted complex – a likely explanation for why its role is so influential on what happens next in the signaling pathway.

“The aggregation of these proteins means they need each other – and this is an exciting finding,” said Liu, also co-director of the Viruses and Emerging Pathogens program in Ohio State’s Infectious Diseases Institute. “A big complex like this can recruit additional molecules, enhancing the efficiency of the signal transduction pathway.”

These recruited molecules are part of a cascade of messages that lead to type I interferon production and NF-kB inflammatory cytokines from virus-infected cells, both of which play a crucial role in limiting viral infection in the early stage. In experiments utilizing multiple types of human and animal cell cultures, this mechanism was seen to inhibit infection of cells by HIV, Ebola and Zika viruses.

In contrast, when cells were modified so they didn’t produce the SERINC5 protein, these viruses took over cells and replicated more – suggesting how important SERINC’s role is in the process that potentiates the primary antiviral response of host cells.

This work was conducted in two members of the SERINC family, SERINC3 and SERINC5, but Liu speculated that other versions may have similar effects. But there is still a lot to learn – such as what triggers SERINC5 to relocate itself from the cell surface to mitochondria, and what its normal cellular functions are when there is no virus around.

As for whether the protein’s functions can suppress the COVID-19 virus, Liu is optimistic.

“My speculation is the SERINC protein would have an effect because we know type I interferons are so important to controlling a virus in the early stage, and this molecule can enhance the type I interferon response,” he said. “Finding one protein that can modulate a key pathway that has a broad antiviral effect – well, you can imagine this protein has a broad antiviral function.”

This work was supported by the National Institutes of Health and the National Cancer Institute (NCI).

Co-authors include Cong Zeng, Tianliang Li, Jingyou Yu, Yi-Min Zheng, Jacob Yount and Haitao Wen, all of Ohio State, and Abdul Waheed and Eric Freed of the NCI HIV Dynamics and Replication Program.

The strategy involves dampening the activity of an enzyme produced by some white blood cells when they’re responding to the vaccine challenge. When highly active, this enzyme breaks down not just the pathogen – its job – but also degrades pieces of cells that participate in the immune response.

Research in mice showed that an experimental COVID-19 vaccine containing a compound to inhibit the enzyme stimulated a robust antibody response that included immunity in the nose and mouth, ultimately providing extra protection for airways and the gastrointestinal tract.

“Our approach is to improve ‘border control.’ The benefits are broad because in addition to providing protection in the bloodstream like most vaccines do, we also have excellent protection in the doors and windows of the body that communicate with the outside,” said senior study author Prosper Boyaka, professor and chair of the Department of Veterinary Biosciences at The Ohio State University.

“If we protect the mucosal area where the pathogen enters, then even if you don’t reach total immunity there, you limit the amount of pathogen that enters the body so the antibodies inside are more efficient at clearing the infection.”

The experimental vaccine was produced by packaging a segment of the SARS-CoV-2 (the virus that causes COVID-19) spike protein as an antigen with the common vaccine ingredient aluminum salts and an enzyme inhibitor. The findings suggest this affordable design could be particularly helpful in developing countries, where cold storage needed for existing vaccines is a challenge, said Boyaka, also an investigator and program director in Ohio State’s Infectious Diseases Institute.

The study was published online Aug. 5 in Proceedings of the National Academy of Sciences.

There is an irony to the use of aluminum salts (also known as alum) in about 70% of the world’s vaccines: While alum’s presence actually enhances the immune response, it also recruits the white blood cells that secrete the enzyme, called elastase.

Alum is inexpensive to obtain or produce and can be stored at room temperature, and is effective at promoting development of a bloodstream-based antibody response to vaccination. But it doesn’t do much for cell-mediated immunity that improves protection against viruses and bacteria that use cells to reproduce, and can’t generate a useful number of antibodies in the body’s portals of entry for most pathogens: the nose, mouth and genitourinary tract.

The researchers found that suppressing elastase in a vaccine containing alum had the dual benefits of broadening and speeding up the antibody response in the bloodstream and triggering the specific types of antibodies needed for immune protection of mucous membranes.

“We found a way to have the cells come and help the immune response to develop and the enzyme to break down the pathogen, but we don’t want that response to be so high that it goes out of control. So we’re just putting a brake on the activity those enzymes would have,” Boyaka said. “And we found if you apply that strategy, you can induce a response in the airways even if the vaccine is not given through the airway.”

The experimental vaccine enhanced the magnitude of mouse antibodies, which reacted to the same section of the spike protein in the vaccine that antibodies in plasma from COVID-19 patients attach to, as well as generating antibodies in mucosal areas. Immunized mice lacking the gene for the enzyme developed high-affinity antibodies as well.

To further test the concept, the researchers found the enzyme-suppressing compound used in the study triggered production of specialized inflammation-regulating cells in cultures of human immune cells and pig spleen cells, showing that this strategy could improve vaccine immune responses in other species – including people.

Boyaka’s team envisions that a future injected vaccine containing an elastase inhibitor could expand SARS-CoV-2 vaccination availability across the world and even be used to boost existing vaccines.

“COVID will stay with us for some time, unfortunately, with the new variants,” he said. “What we need to do is have a portfolio of options that we could use depending on the health environment.

“Reprogramming the immune response induced by an injected vaccine containing alum is a way to make the vaccine more efficient for what we need. This could be a cheap and simple approach that can benefit people in developing countries.”

This work was supported by grants from the National Institutes of Health and an Ohio State Office of Research COVID-19 seed grant. A patent application has been filed spanning this research; the overall patent portfolio includes an additional U.S. issued patent.

Co-authors, all from Ohio State, include Eunsoo Kim, Zayed Attia, Rachel Woodfint, Cong Zeng, Sun Hee Kim, Haley Steiner, Rajni Kant Shukla, Namal Liyanage, Shristi Ghimire, Jianrong Li, Gourapura Renukaradhya, Abhay Satoskar, Amal Amer, Shan-Lu Liu and Estelle Cormet-Boyaka.

In the study, scientists defined the atomic-level architecture of the connection between a protein on the surface of a parasite-infected red blood cell when it binds to a receptor on the surface of an immune cell.

When that protein-receptor connection is made under normal circumstances, the infected red blood cell, hijacked by the disease-causing parasite, de-activates the immune cell – meaning the body won’t fight the infection. A drug designed to fit into that space could block the interaction, allowing the immune system to get to work clearing away the pathogen.

In a previous study, a team including the Ohio State University and National Institutes of Health scientists who led this research did similar work with another immune cell receptor that the protein, called RIFIN, binds to in its bid to suppress the immune response.

Through a genome-wide analysis of the parasite that causes malaria, the scientists found RIFIN exerts the same type of immune-suppressing function in various species of Plasmodium infecting humans, gorillas and chimpanzees. This suggests it is a mechanism that has not changed over the course of evolution – meaning this function is critical to the parasite’s success and therefore an attractive target for intervention.

The researchers envision either a vaccine or a chemical compound, or both, could be developed to disable this function, reducing the risk of severe malaria cases that require hospitalization and rapid treatment.

“RIFIN targets two receptors to down-regulate immune function so the parasite can evade immune surveillance and survive. If we can lift the immunosuppression, the human immune system can take care of the rest,” said Kai Xu, assistant professor of veterinary biosciences at Ohio State and co-lead author of the study. “Inhibition of the immune response is one of the major reasons severe malaria infection is so hard to deal with.”

Xu co-led the research with Peter Kwong of the National Institute of Allergy and Infectious Diseases (NIAID) Vaccine Research Center.

The study is published in Nature Communications.

This work focuses on species of Plasmodium that cause the worst cases of malaria – many of the 200 million people infected annually have mild symptoms, but severe cases can cause respiratory distress and organ failure. More than 400,000 people die of the disease each year. There are several drugs used to treat malaria, but current medications are losing effectiveness because the parasites have developed resistance to them.

Humans are infected through the bite of a mosquito carrying the parasite. Once in the human body, the parasites transform themselves in the liver so they can then infect red blood cells, reproduce and release toxic factors, which leads to clinical symptoms of disease.

The members of the RIFIN family of parasitic proteins – of which there are 200-plus – can do lots of things to exacerbate the infection once the parasite has reached red blood cells. A small subset of them bind to two receptors, LAIR1 and LILRB1, on B-, T- and NK cell surfaces to keep those immune cells dormant.

Capturing the protein-receptor interactions with X-ray crystallography in enough detail to define the precise structure at the binding site can be tricky because they happen so quickly and are dynamic. The researchers observed the connections as they naturally happen, but a bit of serendipity provided them with an even better option. It turns out that antibodies induced in some people who have had malaria contain genes from the LAIR1 receptor, and by being part of a parasite-specific antibody, the LAIR1 segment develops a very high attraction to RIFIN. Using those unusual antibody structures to observe the LAIR1 segment’s attachment to RIFIN gave the team a much, much closer look at the structure of their bond.

From here, the researchers plan to focus their efforts on the 20 or so RIFIN family members that are attracted to and bind with the two immune cell receptors.

“RIFIN is a large and diverse parasitic protein family. However, the subset of RIFIN molecules that bind to LAIR1 and LILRB1 is less diversified and shares common features, so we only focus on that small subset,” Xu said. “We want to generate a drug that can specifically target the receptor-binding interface on RIFIN, blocking one of the important immune escape mechanisms of the parasite. That’s the future direction.”

This work was funded by the NIAID Vaccine Research Center, a GenScript Innovation grant and the Frederick National Laboratory for Cancer Research. The researchers also used the U.S. Department of Energy Office of Science’s Advanced Photon Source.

In lab studies, researchers identified two groups of molecules that were effective against virus variants. Using different mechanisms, nanobodies in each group bypassed mutations and disabled the virus’s ability to bind to the receptor that lets it enter host cells.

Though vaccination is enabling the resumption of some pre-pandemic activities in parts of the world, SARS-CoV-2 is rapidly working its way around vaccines by mutating itself. In this study, the nanobodies neutralized three emerging variants: Alpha, Beta and Gamma.

“Companies have already started introducing the variants of concern into the construct of booster shots of the existing vaccines,” said Kai Xu, assistant professor of veterinary biosciences at The Ohio State University and a co-lead author of the research. “But the virus is constantly mutating, and the speed of mutation may be faster than we can capture. Therefore, we need to utilize multiple mechanisms to control the virus spread.”

An accelerated article preview of the study is published online in Nature.

Nanobodies are antibodies derived from immunization of camelid mammals – such as camels, llamas and alpacas – that can be re-designed into tiny molecules that mimic human antibody structures and functions.

For this work, the researchers immunized llamas to produce single-chain antibodies against SARS-CoV-2. They also immunized “nanomice,” transgenic mice with a camelid gene that had been engineered by research fellow Jianliang Xu in the lab of Rafael Casellas, senior investigator at the National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS), to generate nanobodies similar to those produced by camelids.

The team enhanced the nanobodies’ power by immunizing the animals first with the receptor binding domain (RBD), a part of the viral surface spike protein, and following with booster shots containing the entire spike protein.

“By using this sequential immunization strategy, we generated nanobodies that can capture the virion by recognizing the receptor binding domain with very high affinity,” Xu said.

The scientists tested different nanobodies’ neutralization capacity, mapping the surface of the RBD, conducting functional and structure analyses, and measuring the strength of their affinity to narrow the candidate molecules from a large library to six.

The coronavirus is highly infectious because it binds very tightly to the ACE2 receptor to gain access to lung and nasal cavity cells in humans, where it makes copies of itself to infect other cells. The receptor binding domain on the spike protein is fundamental to its success in attaching to ACE2.

“That RBD-ACE2 interface is on the top of the receptor binding domain – that region is the primary target for the protective human antibodies, generated by vaccination or previous infection, to block the viral entry,” Xu said. “But it is also a region frequently mutated in the variants.”

The way mutants have emerged so far suggests long-term reliance on current vaccines will eventually be compromised, the researchers say, because antibody effectiveness is affected significantly by those mutants at the interface.

“We found that certain nanobodies can recognize a conserved region of the receptor binding domain, a hidden location that is too narrow for human antibodies to reach,” Xu said. And attaching at this location, even though it is some distance away from where RBD connects to ACE2, still accomplishes what is intended – blocking SARS-CoV-2 from entering a host cell.

The other group of nanobodies, attracted to the RBD-ACE2 interface, while in their original form could not neutralize certain variants. However, when the researchers engineered this group to be homotrimers – three copies linked in tandem – the nanobodies achieved potent neutralization of the virus. Altering the structure of the nanobodies that attached to the conserved region of RBD in the same way enhanced their effectiveness as well.

There is much more research ahead, but the findings suggest nanobodies could be promising tools to prevent COVID-19 mortality when vaccines are compromised, Xu said.

“Our future plan is to further isolate antibodies specifically against emerging variants for therapeutic development, and to find a better solution for vaccines by learning from those antibodies,” he said.

An HIV vaccine researcher at the NIH before joining Ohio State, Xu collaborated with multiple labs on this research. Jianliang Xu and Rafael Casellas of NIAMS, and Peter Kwong of the National Institute of Allergy and Infectious Diseases, are also equal contributors on the study. In addition to numerous NIH agencies, co-author institutions include Rockefeller University, the Aaron Diamond AIDS Research Center at Columbia University and the Frederick National Laboratory for Cancer Research.

This work was supported by NIAMS, the National Cancer Institute, NIH Helix Systems, NIAID and the Frederick National Laboratory for Cancer Research.

The 35,000-square-foot state-of-the-art building provides a rare opportunity to advance clinical training for Ohio State’s College of Veterinary Medicine students. The goal of this “spectrum of care” educational model is to ensure that veterinary graduates entering private practice have the competence and confidence to provide more in-clinic treatment and to offer a variety of treatments for pets that belong to pet parents across a wide-ranging socioeconomic spectrum.

Ohio State veterinary medicine students will gain knowledge and mastery of a broad array of diagnostic, therapeutic, business and communication skills. This change to the college’s educational model was driven by input and feedback from faculty, students, practitioners and alumni. 

“The partnership between Ohio State and the Stanton Foundation runs deep,” said President Kristina M. Johnson during the clinic’s ribbon cutting ceremony.

“Sparked by its namesake 90 years ago, this relationship is anchored in our shared mission of uplifting those around us. And it is helping to transform the way our students learn, the environments in which they train and the opportunities they have to hone their skills – the benefits of which directly support our communities by helping countless pets and companion animals.”

The college has engaged veterinary partners across the state and beyond in the development of this concept since planning began, and those partners will also be actively involved in the coaching of students – another way the college hopes to provide a much broader private practice experience for students, ultimately producing practice-ready graduates who are highly sought-after by employers upon graduation. 

Traditionally, veterinary students at Ohio State have received much of their clinical training primarily by observing complex medical and surgical cases evaluated and treated in the Veterinary Medical Center by specialists using sophisticated and extensive diagnostic tests, imaging studies and treatment procedures. Although this traditional educational model contributes significantly to a student’s knowledge of how to manage cases, weaving the spectrum of care philosophy into each phase of the students’ four-year DVM curriculum emphasizes hands-on clinical opportunities across a broad range of cases, including more training in business, effective communication skills and other non-technical competencies. 

Few veterinary colleges provide an immersive clinical skills training program combined with a hands-on general practice program.

Approximately 80% of Ohio State’s veterinary medicine graduates enter general practice. The opening of the clinic provides an opportunity for every student to gain a breadth of experience in a realistic general practice setting starting in their first semester and culminating in their capstone clinical training during their fourth year. Students will be involved in all aspects of patient care and client service at the new facility and will be coached by faculty who are experienced private practitioners.

“The spectrum of care concept aims to address the growing problem of affordability of veterinary care by providing a continuum of acceptable care that considers available evidence-based medicine while remaining responsive to client expectations and financial limitations, thereby successfully serving an economically diverse clientele,” said Rustin Moore, dean of the College of Veterinary Medicine and Ruth Stanton Chair in Veterinary Medicine. 

The Frank Stanton Veterinary Spectrum of Care Clinic is funded by a gift from the Stanton Foundation, which has supported the college’s goal to prepare even more confident and competent graduates for several years. A transformational gift in 2016 made it possible for the college to construct a state-of-the-art veterinary clinical and professional skills center, expand summer student externships and hire additional faculty and staff. 

“Frank Stanton was the longtime president of CBS, creator of the Kennedy-Nixon debates, lifelong dog lover and a proud alumnus of The Ohio State University. After his Corgi, Foxo, received critical care at a prominent east coast veterinary hospital, he realized that many pet owners could never afford the care that Foxo had received,” said Steve Kidder, spokesperson for the Stanton Foundation. 

“As an astute businessman, he also recognized that philanthropy alone could not solve this problem. He thus began a search for a more imaginative and sustainable approach that would help millions, not thousands, of animals. The Stanton Foundation is gratified that it has found in The Ohio State University College of Veterinary Medicine a partner to begin translating his vision into reality.”

Added Roger B. Fingland, executive director and chief medical officer of the Veterinary Health System and the Frank Stanton Chair, “The Stanton Foundation has joined us along this path in almost every step, because visions and dreams aligned. This would not have happened without that alignment.

“We cannot even begin to adequately express our gratitude for what the Stanton Foundation has done to change the lives of so many pets and people – but what we can say with great humility, the utmost confidence, and with profound appreciation to the Stanton Foundation, is that Frank Stanton changed the world.”

The infection is called human monocytic ehrlichiosis, and is one of the most prevalent and potentially life-threatening tick-borne diseases in the United States. The disease initially causes flu-like symptoms common to many illnesses, and in rare cases can be fatal if left untreated.

Most antibiotics can’t build up in high enough concentrations to kill the infection-causing bacteria, Ehrlichia chaffeensis, because the microbes live in and multiply inside human immune cells. Commonly known bacterial pathogens like Streptococcus and E. coli do their infectious damage outside of hosts’ cells.

Ohio State University researchers created nanobodies intended to target a protein that makes E. chaffeensis bacteria particularly infectious. A series of experiments in cell cultures and mice showed that one specific nanobody they created in the lab could inhibit infection by blocking three ways the protein enables the bacteria to hijack immune cells.

“If multiple mechanisms are blocked, that’s better than just stopping one function, and it gives us more confidence that these nanobodies will really work,” said study lead author Yasuko Rikihisa, professor of veterinary biosciences at Ohio State.

The study provided support for the feasibility of nanobody-based ehrlichiosis treatment, but much more research is needed before a treatment would be available for humans. There is a certain urgency to coming up with an alternative to the antibiotic doxycycline, the only treatment available. The broad-spectrum antibiotic is unsafe for pregnant women and children, and it can cause severe side effects.

“With only a single antibiotic available as a treatment for this infection, if antibiotic resistance were to develop in these bacteria, there is no treatment left. It’s very scary,” Rikihisa said.

The research is published this week in Proceedings of the National Academy of Sciences.

The bacteria that cause ehrlichiosis are part of a family called obligatory intracellular bacteria. E. chaffeensis not only requires internal access to a cell to live, but also blocks host cells’ ability to program their own death with a function called apoptosis – which would kill the bacteria.

“Infected cells normally would commit suicide by apoptosis to kill the bacteria inside. But these bacteria block apoptosis and keep the cell alive so they can multiply hundreds of times very rapidly and then kill the host cell,” Rikihisa said.

A longtime specialist in the Rickettsiales family of bacteria to which E. chaffeensis belongs, Rikihisa developed the precise culture conditions that enabled growing these bacteria in the lab in the 1980s, which led to her dozens of discoveries explaining how they work. Among those findings was identification of proteins that help E. chaffeensis block immune cells’ programmed cell death.

The researchers synthesized one of those proteins, called Etf-1, to make a vaccine-style agent that they used to immunize a llama with the help of Jeffrey Lakritz, professor of veterinary preventive medicine at Ohio State. Camels, llamas and alpacas are known to produce single-chain antibodies that include a large antigen binding site on the tip.

The team snipped apart segments of that binding site to create a library of nanobodies with potential to function as antibodies that recognize and attach to the Etf-1 protein and stop E. chaffeensis infection.

“They function similarly to our own antibodies, but they’re tiny, tiny nano-antibodies,” Rikihisa said. “Because they are small, they get into nooks and crannies and recognize antigens much more effectively.

“Big antibodies cannot fit inside a cell. And we don’t need to rely on nanobodies to block extracellular bacteria because they are outside and accessible to ordinary antibodies binding to them.”

After screening the candidates for their effectiveness, the researchers landed on a single nanobody that attached to Etf-1 in cell cultures and inhibited three of its functions. By making the nanobodies in the fluid inside E. coli cells, Rikihisa said her lab could produce them at an industrial scale if needed – packing millions of them into a small drop.

She collaborated with co-author Dehua Pei, professor of chemistry and biochemistry at Ohio State, to combine the tiny molecules with a cell-penetrating peptide that enabled the nanobodies to be safely delivered to mouse cells.

Mice with compromised immune systems were inoculated with a highly virulent strain of E. chaffeensis and given intracellular nanobody treatments one and two days after infection. Compared to mice that received control treatments, mice that received the most effective nanobody showed significantly lower levels of bacteria two weeks after infection.

With this study providing the proof of principle that nanobodies can inhibit E. chaffeensis infection by targeting a single protein, Rikihisa said there are multiple additional targets that could provide even more protection with nanobodies delivered alone or in combination. She also said the concept is broadly applicable to other intracellular diseases.

“Cancers and neurodegenerative diseases work in our cells, so if we want to block an abnormal process or abnormal molecule, this approach may work,” she said.

This study was supported by the National Institutes of Health.

Additional co-authors, all from Ohio State, include Wenqing Zhang, Mingqun Lin, Qi Yan, Khemraj Budachetri, Libo Hou, Ashweta Sahni, Hongyan Liu and Nien-Ching Han.

Tests in mice infected with high doses of influenza showed that the treatment could improve lung function in very sick mice and prevent progression of disease in mice that were pre-emptively treated after being exposed to the flu.

The hope is that it may also help humans infected with the flu, and potentially other causes of acute respiratory distress syndrome (ARDS) such as SARS-CoV-2 infection.

Specific cells in mice are less able to make key molecules after influenza invades the lungs, reducing their ability to produce a substance called surfactant that enables lungs to expand and contract. The shortage of surfactant is linked to ARDS, an illness so serious that it typically requires mechanical ventilation in an ICU.

Researchers bypassed the blocked process in mice by re-introducing the missing molecules alone or in combination as an injected or oral treatment. The results: normalized blood oxygen levels and reduced inflammation in mouse lungs – effects that could make a person well enough for hospital discharge.

“The most important and impressive thing in this study is the fact that we have benefits even when we treat late in the disease process. If we could develop a drug based on these findings, you could take somebody who’s going to have to go on a ventilator and stop that completely,” said Ian Davis, professor of veterinary biosciences at The Ohio State University and senior author of the study. “There’s nothing out there now that can do this for ARDS that will bring them back to that degree, and certainly not for flu.”

ARDS can also result from infections, cancer, trauma and many other ailments. Though this therapy has been tested in the context of the flu, Davis said its reliance on fixing a broken cell function in the host rather than killing the virus suggests it has potential to treat virtually any lung injury.

The study was published online recently in the American Journal of Respiratory Cell and Molecular Biology.

The experimental treatment consists of molecules known as liponucleotides, which are essential for making surfactant in the lungs. Davis analyzed lung cells from flu-infected mice and determined that the pathway to surfactant production was disrupted, with one of the two necessary liponucleotides completely undetectable.

“The thinking before was that the reason there was less surfactant in mice with flu-related ARDS was because cells are dying. This defect is in some ways better – if cells are dying, there’s not much you can do, but if there’s a problem with the cell’s metabolism, maybe you can fix it,” Davis said.

And fix it – in mice – he did, developing therapies containing the missing liponucleotide molecule alone or combined with one or two others.

Davis and colleagues inoculated mice with high doses of H1N1 influenza and then treated some mice with liponucleotides once daily for five days and others just a single time five days after exposure. The mice receiving daily treatment were protected from getting seriously ill, and the very sick mice treated on the fifth day, whose severe blood-oxygen loss and lung inflammation had cause ARDS, showed significant improvement.

“Obviously that’s what you need in someone with severe influenza – we want to take someone who is already in the ICU and help them get out faster, or head going to the ICU off at the pass,” Davis said.

Liponucleotides don’t kill the flu virus – which is the point.

“I’ve always been interested in finding new therapies to treat lung injury,” he said. “The problem with anti-viral drugs is you probably need a different drug for every virus. Also, many viruses can quickly mutate to become resistant to these drugs.

“Our approach is to fix the patient. Once the virus has caused the injury – the inflammation – it doesn’t really matter if it stays or goes away.”

There’s still a lot to learn. The agents have a strong anti-inflammatory effect, but don’t fully restore the surfactant-production process – and Davis isn’t sure why that is. Studies so far have been based on findings in a single type of lung cell, but the scientists haven’t confirmed that those cells are the ones responding to the therapy – any number of other cells in the immune system, blood vessels or heart could also play a role.

Despite the unknowns, Davis said that because the missing liponucleotides naturally exist in mammals, including humans, they are considered safe and unlikely to cause side effects, even if they go unused in the body.

The Ohio State Innovation Foundation has filed patents covering the scope of Davis’ discoveries, which may also extend to patients suffering from other forms of lung damage that cause inflammation and a drop in blood oxygen levels.

This work was supported by the National Heart, Lung and Blood Institute.

Co-authors include Lucia Rosas, Lauren Doolittle, Lisa Joseph, Hasan El-Musa and Judy Hickman-Davis of Ohio State’s College of Veterinary Medicine; Michael Novotny from the Cleveland Clinic; and Duncan Hite from the University of Cincinnati.

Since 2009, GOHi has been on the front lines in Ethiopia working with in-country partners to strengthen capacity using a One Health approach — one that brings together multiple disciplines working globally to address the spread of disease, promote health and emphasize the connection among humans, animals, plants and the environment.

As new diseases emerge, the need for health system preparedness across the globe is vital for nations to prevent spread of pathogens, detect and report epidemics, and respond to and mitigate the spread of those epidemics. The current COVID-19 pandemic urgently underscored these needs. Under-preparedness in one country is a global risk to all.

“This award exemplifies the critical importance of focusing on public health, which is a global issue and very timely,” said Grace Wang, executive vice president for research, innovation and knowledge. “By harnessing the capability of Ohio State’s world-class research faculty, we are working to find innovative solutions to global challenges and are pleased to partner with the Centers for Disease Control and Prevention and Ethiopia’s public health system.”

With this award, GOHi aims to address three specific focus areas in order to achieve International Health Regulation standards and benchmarks; strengthen surveillance, laboratory and workforce capacity; improve data management; and develop a well-linked response network for disease outbreaks and public health emergencies.

“Right now, the world needs a major effort to strengthen surveillance, laboratory and workforce capabilities,” said Wondwossen Gebreyes, GOHi executive director. “The GOHi consortium on campus along with global partners are committed to tackling the world’s toughest health challenges at the interface of humans, animals, plants and the environment, including COVID-19. The impact of this work will save lives while we continue to fight this pandemic. It will also have a lasting impact on prevention and control of future zoonotic viral and drug resistant bacterial infections among others.”

GOHi, with participation from Ohio State’s College of Medicine, College of Public Health and College of Veterinary Medicine and additional faculty support from the College of Arts and Sciences and College of Food, Agricultural, and Environmental Sciences, will partner with the Ethiopian Public Health Institute and in-country Regional Health Bureaus to accomplish the project.

“It is very exciting to receive this award, which has come at the right time, when we need it the most,” said Getnet Yimer, regional director, Global One Health Eastern Africa Office. “We are committed to continue working with all partners and the government agencies to sustain our gains and achieve beyond the planned milestones.”

Over the next five years, the project will expand the number of laboratories with the ability to test and report Influenza-like illnesses and severe acute respiratory illnesses, like COVID-19; train health professionals and laboratory staff on better specimen collection and transportation techniques; improve data collection and reporting and implementation of appropriate mitigation measures for severe respiratory illnesses based on that data; support equipment procurement; and improve quality management throughout the laboratory network to ensure consistent, reliable quality testing.

The ultimate goal will be to enhance and expand the Ethiopian public health system to more comprehensively and efficiently manage the multiple elements that contribute to epidemics of global (national and international) concern.

This project serves as an example of institutional teamwork that advances the university’s commitment as a global institution, engaging in meaningful and beneficial partnerships to gain and share knowledge and find sustainable solutions to the world’s most complex issues.

Scientists attribute the vaccine candidate’s effectiveness to strategic production of the antigen to stimulate immunity: using a specific snippet of the coronavirus spike protein gene, and inserting it into a sweet spot in the measles vaccine genome to boost activation, or expression, of the gene that makes the protein.

Even with several vaccines already on the market, researchers say this candidate may have advantages worth exploring – especially related to the measles vaccine’s established safety, durability and high-efficacy profile.

“The measles vaccine has been used in children since the 1960s, and has a long history of safety for children and adults,” said Jianrong Li, senior author of the study and a professor of virology in The Ohio State University Department of Veterinary Biosciences.

“We also know the measles vaccine can produce long-term protection. The hope is that with the antigen inside, it can produce long-term protection against SARS-CoV-2. That would be a big advantage, because right now we don’t know how long protection will last with any vaccine platforms.”

The Ohio State Innovation Foundation has exclusively licensed the technology to Biological E. Limited (BE), a Hyderabad, India-based vaccine & pharmaceutical company.

The research is published online today (March 9, 2021) in the journal Proceedings of the National Academy of Sciences.

The coronavirus that causes COVID-19 uses the spike protein on its surface to bind to its target cells in the nose and lungs, where it makes copies of itself and releases them to infect other cells. Like all vaccines, this candidate initiates the production of antibodies that recognize the new protein as foreign, training the immune system to attack and neutralize the spike protein if SARS-CoV-2 ever enters the body.

Li created the COVID-19 vaccine using a live attenuated measles virus as a vehicle with colleagues Mijia Lu, a postdoctoral researcher in Li’s laboratory and first author on the paper, and co-authors Stefan Niewiesk, Ohio State professor of veterinary biosciences, and Mark Peeples, professor of pediatrics at Ohio State and a researcher at Nationwide Children’s Hospital in Columbus.

For this work, the researchers tested seven versions of the spike protein to find the most effective antigen. They landed on a stabilized “prefusion” version of the protein – the shape the protein is in before it infects a cell.

The scientists inserted the prefusion spike protein gene containing manufacturing instructions into a segment of the measles vaccine genome to generate high expression of the protein, reasoning that the more SARS-CoV-2 spike protein produced, the better the immune response.

The team tested the vaccine candidate in several animal models to gauge its effectiveness, and found that the vaccine induced high levels of neutralizing antibodies against SARS-CoV-2 in all of the animals.

Some may think most humans’ immunity to measles, thanks to decades of widespread vaccination, would render its status as a coronavirus vaccine vehicle useless. To quell those concerns, researchers gave cotton rats a measles vaccine and showed that a second immunization with the measles-based SARS-CoV-2 vaccine candidate could induce a strong neutralizing antibody response to the coronavirus.

Genetically modified mice produced helper T cells – a type of white blood cell – in response to the vaccine, another important way the body fights infection, and severe disease in particular.

“The orientation of T helper cells induced by a vaccine is an important predictor of protection, and this vaccine mainly induces Th1 cells, which enhances the safety and efficacy of the vaccine,” said co-author Amit Kapoor, associate professor of pediatrics at Ohio State and a researcher at Nationwide Children’s Hospital.

Golden Syrian hamsters, which are susceptible to contracting COVID-19, received the vaccine and were then injected with the coronavirus. The vaccinated hamsters were protected from lung infection and other sickness symptoms indicated through weight loss.

“When we looked at the amount of neutralizing antibody induced in the hamster, it was actually higher than from people who had been infected with COVID, suggesting the vaccine may be better than SARS-CoV-2 infection in inducing protective immunity. That was our goal,” Peeples said.

The researchers have confidence in the platform not only because the measles vaccine is safe, effective and affordable to produce, but because several experimental measles-based vaccines against other viruses are in development. A vaccine against chikungunya virus, spread by mosquitos, has been shown to be safe, well-tolerated and good at provoking an immune response in a Phase 2 clinical trial.

And even with a variety of COVID-19 vaccines now available in the United States and other countries, there is still a lot to learn about which are the safest and most effective for specific populations, such as children and pregnant women, and which vaccines are the most economical to produce.

“We can make vaccines much more quickly now than in the past. But if we had to do it the traditional way this time, we wouldn’t have a vaccine protecting us in this short amount of time,” Niewiesk said. “The mRNA vaccines in use now were made in record time. And they protect against disease and are safe. Although not quite as fast, we were able to make this vaccine much more quickly than the original measles vaccine.

“We don’t yet know how long the mRNA vaccines will protect or how much they will cost. In the meantime, an alternative vaccine that should protect for a long time, is easy to manufacture and cheap seems like a good idea.”

This study was supported by startup funds and bridge funds from Ohio State’s Department of Veterinary Biosciences and College of Veterinary Medicine, a seed grant from Nationwide Children’s Hospital and grants from the National Institutes of Health.

Additional co-authors are Yuexiu Zhang, Anzhong Li, Olivia Harder, Cong Zeng, Xueya Liang, Shan-Lu Liu and Prosper Boyaka of Ohio State’s Department of Veterinary Biosciences; Piyush Dravid, Sheetal Trivedi, Mahesh KC, Supranee Chaiwatpongsakorn, Masako Shimamura, Asuncion Mejias and Octavio Ramilo of the Research Institute at Nationwide Children’s Hospital; and Ashley Zani, Adam Kenney, Chuanxi Cai and Jacob Yount of the Department of Microbial Infection and Immunity in Ohio State’s College of Medicine.

Scientists have developed protein fragments – called peptides – that fit snugly into a groove on the SARS-CoV-2 Spike protein that it would normally use to access a host cell. These peptides effectively trick the virus into “shaking hands” with a replica rather than with the actual protein on a cell’s surface that lets the virus in.

Previous research has determined that the novel coronavirus binds to a receptor protein on a target cell’s surface called ACE2. This receptor is located on certain types of human cells in the lung and nasal cavity, providing SARS-CoV-2 many access points to infect the body.

For this work, Ohio State University scientists designed and tested peptides that resemble ACE2 enough to convince the coronavirus to bind to them, an action that blocks the virus’s ability to actually get inside the cell.

“Our goal is that any time SARS-CoV-2 comes into contact with the peptides, the virus will be inactivated. This is because the virus Spike protein is already bound to something that it needs to use in order to bind to the cell,” said Amit Sharma, co-lead author of the study and assistant professor of veterinary biosciences at Ohio State. “To do this, we have to get to the virus while it’s still outside the cell.”

The Ohio State team envisions delivering these manufactured peptides in a nasal spray or aerosol surface disinfectant, among other applications, to block circulating SARS-CoV-2 access points with an agent that prevents their entry into target cells.

“With the results we generated with these peptides, we are well-positioned to move into product-development steps,” said Ross Larue, co-lead author and research assistant professor of pharmaceutics and pharmacology at Ohio State.

The study is published in the January issue of the journal Bioconjugate Chemistry.

SARS-CoV-2, like all other viruses, requires access to living cells to do its damage – viruses hijack cell functions to make copies of themselves and cause infection. Very rapid virus replication can overwhelm the host system before immune cells can muster an effective defense.

One reason this coronavirus is so infectious is because it binds very tightly to the ACE2 receptor, which is abundant on cells in humans and some other species. The Spike protein on the SARS-CoV-2 surface that has become its most recognizable characteristic is also fundamental to its success in attaching to ACE2.

Recent advances in crystallizing proteins and microscopy have made it possible to create computer images of specific protein structures alone or in combination – such as when they bind to each other.

Sharma and his colleagues closely examined images of the SARS-CoV-2 Spike protein and ACE2, zooming in on precisely how their interactions occur and what connections are required for the two proteins to lock into place. They took notice of a spiral ribbon-like tail on ACE2 as the focal point of the attachment, which became the starting point for designing peptides.

“Most of the peptides we designed are based on the ribbon contacting the Spike,” said Sharma, who also holds a faculty appointment in microbial infection and immunity. “We focused on creating the shortest possible peptides with the minimum essential contacts.”

The team tested several peptides as “competitive inhibitors” that could not only securely bind with SARS-CoV-2 Spike proteins, but also prevent or lower viral replication in cell cultures. Two peptides, one with the minimum contact points and another larger one, were effective at reducing SARS-CoV-2 infection in cell studies compared to controls.

Sharma described these findings as the beginning of a product-development process that will be continued by the team of virologists and pharmaceutical chemists collaborating on this work.

“We are taking a multipronged approach,” Sharma said. “With these peptides, we have identified the minimal contacts needed to inactivate the virus. Going forward we plan to focus on developing aspects of this technology for therapeutic purposes.

“The goal is to neutralize the virus effectively and potently, and now, because of the emergence of variants, we’re interested in assessing our technology against the emerging mutations.”

This research was supported by Ohio State institutional startup funds.

Sharma and Larue are inventors on a provisional patent application covering this technology. Other co-authors, from Ohio State’s colleges of Pharmacy, Medicine and Veterinary Medicine, are Enming Xing, Adam Kenney, Yuexiu Zhang, Jasmine Tuazon, Jianrong Li, Jacob Yount and Pui-Kai Li.

Graphic generated using ChimeraX software and published protein structure.

This year’s honorees are scientists in the colleges of Arts and Sciences, Medicine and Veterinary Medicine.

The AAAS Fellowship, recognizing scientifically or socially distinguished efforts to advance science or its applications, is one of the most prestigious honors a U.S. scientist can receive. Fellows are elected by their academic peers.

“AAAS Fellows are honored for achievements in a specific discipline and dedication to the principle of improving the world through science,” Ohio State President Kristina M. Johnson said. “These faculty embody what Ohio State’s research enterprise is all about – making an impact through scholarship.”

Ohio State’s newest Fellows are:

Janet Box-Steffensmeier, professor of political science. For distinguished contributions to political methodology, including duration analysis and time series, and service to the advancement of political methodology and study of American politics.

Elizabeth Cooksey, professor of sociology. For distinguished contributions to the field of social demography, with specializations in life course transitions, adolescent sexual behavior, and the development of youth and children.

Debbie Guatelli-Steinberg, professor of anthropology. For distinguished contributions to biological anthropology, particularly using dental hard tissues to advance understanding of growth and biological relationships in non-human primates and fossil hominins.

Yuri Kovchegov, professor of physics. For foundational contributions to the theoretical understanding of parton saturation effects in quantum chromodynamics and their manifestation in high-energy collisions with strongly interacting particles.

Zhengyu Liu, professor of geography. For distinguished contributions to our understanding of climate dynamics, particularly in pioneering data-model experiments to explore the physics of present and past climate variability.

Shan-Lu Liu, professor of veterinary biosciences. For distinguished contributions to our understanding of virus-host interaction and viral pathogenesis, as well as impact on scientific communication, diversity and international collaboration.

Paul Martini, professor of astronomy. For distinguished contributions to the development of astronomical instrumentation, and the evolution of black holes and galaxies.

Jay Myung, professor of psychology. For distinguished applied and basic research on computational cognition, Bayesian cognitive modeling, optimal experimental design, adaptive design optimization, model selection and evaluation, and neural networks.

John Olesik, senior research scientist in earth sciences. For distinguished contributions to the field of analytical chemistry, particularly in optical spectroscopy and mass spectrometry.

Nandini Trivedi, professor of physics. For her contributions to the theoretical understanding of quantum matter, characterized by innovative use of quantum Monte Carlo techniques and close experimental collaborations.

Daniel Wozniak, professor of microbial infection and immunity. For contributions to defining patho-adaptive processes and evolution of bacteria during infection in the context of biofilms and animal models of chronic infection.

New Fellows will be inducted during a virtual ceremony on Feb. 13, 2021.

Scientists at The Ohio State University have developed a new lab testing procedure for the detection of antibodies against SARS-CoV-2 that gives results more quickly than existing assays and specifically identifies so-called “neutralizing” antibodies – those that protect by blocking infection of cells.

“With many assays currently in use, we can detect antibodies, but that doesn’t tell us if they’re neutralizing antibodies. We only know the level of antibodies someone has,” said Shan-Lu Liu, professor in the Ohio State College of Veterinary Medicine’s Department of Veterinary Biosciences and the senior author of a new journal article describing the assay.

“Some antibodies might be protective, some might not be protective, and some might even enhance infection – we know with this type of coronavirus and some other viruses, some antibodies can even do harm,” he said. “Our assay examines whether antibodies are potentially protective, which means they prevent a patient from reinfection and block viral replication. That’s the outcome of infection that we want people to have.”

In analyses of blood samples from several different populations that had tested positive for COVID-19, the researchers found with this new assay that, overall, ICU patients had produced the highest concentration of neutralizing antibodies, and convalescent plasma donors and health care workers had the lowest antibody levels.

“So the more severe the disease, the higher the antibody levels produced. And what this tells us is there is a wide spectrum of different antibody levels after infection,” said Liu, also an investigator in the Center for Retrovirus Research and co-director of the Viruses and Emerging Pathogens Program in Ohio State’s Infectious Diseases Institute. “We’re in a pandemic now, but eventually we’ll be able to see not just how many people were infected, but also the outcome. Our assay could be used to tell whether antibodies have been developed in individuals who have had contacts with SARS-CoV-2.”

The research is published online in the journal JCI Insight.

Liu and colleagues developed what is called a “pseudotype” virus neutralizing antibody assay, in which an HIV vector and core is coated with the SARS-CoV-2 spike protein to detect antibodies against the coronavirus. The team applied a new approach by selecting a different form of light-producing enzyme that can be detected conveniently in culture media containing the virus-infected cells. That choice saved several steps, and time, in the detection process without losing accuracy and sensitivity to the target virus.

Co-first authors Cong Zeng, a postdoctoral researcher, and Jack Evans, a student in the molecular, cellular and developmental biology graduate program, completed the majority of the work to develop the assay.

With the new analytical tool in hand, scientists in the College of Veterinary Medicine collaborated with Gerard Lozanski and others in the College of Medicine to analyze 221 patient blood samples to validate the effectiveness of the assay and verify that the detection test could be scaled up for widespread screening.

The samples were derived from 104 hospitalized COVID-19 patients, 49 of whom were in intensive care; 42 health care professionals who had tested positive for COVID-19; 38 convalescent plasma donors (recovered patients who donated plasma for potential therapeutic use in very sick COVID-19 patients); and 37 control samples from patients who had been hospitalized with respiratory conditions before September 2019.

Results showed that, in general, hospitalized patients – and ICU patients in particular – had the highest concentrations, or titers, of neutralizing antibodies in their systems. However, over 14% of those who had been hospitalized had no or very low levels of antibodies.

Of the health care professionals, 40% were negative for neutralizing antibodies and 36% had low concentrations. And more than half of the convalescent blood donors had concentrations of antibodies that were too low to qualify them as donors for treatment of COVID-19 patients, Liu said.

The assay detected no SARS-CoV-2 antibodies in the samples from people who had been sick with other types of respiratory diseases.

The test accuracy was further validated by verifying in a lab setting that the antibodies detected in the COVID-19 patient blood samples did in fact neutralize the authentic SARS-CoV-2 virus.

It won’t be long before the assay is put to a larger test. The effectiveness, sensitivity and specificity of the assay were important factors in Ohio State’s successful application for a $10 million National Cancer Institute grant awarded last month for studies of the long-term impact of COVID-19 on first responders, health care workers and the general population. Two of the co-authors on the JCI Insight paper will be co-principal investigators of the first-responders study: Eugene Oltz, professor and chair of microbial infection and immunity in the College of Medicine, and Linda Saif, Distinguished University Professor in the College of Veterinary Medicine and the Food Animal Health Research Program in the College of Food, Agricultural and Environmental Sciences.

“We will use this assay to conduct a serological study of first responders in Columbus, among others,” Liu said.

Additional applications could include screening for protective qualities in lab-manufactured monoclonal antibodies designed for therapeutic purposes and neutralizing antibody production in vaccine candidates.

Patent applications have been filed covering the assay and its use in a variety of applications. Inventors named on the applications are Liu, Zeng, Evans, and co-authors Panke Qu and Yi Min Zheng.

“We are pleased to have developed a neutralization assay which provides a sensitive, specific and simple method for determining the neutralizing antibody titers in patient serum or plasma with results being achieved in 24 hours – without requiring access to a biosafety level-3 facility,” Liu said.

“We’d like to make it widely available as quickly as possible because we think it’s a rapid and simple system compared to other systems.”

Additional co-authors, all at Ohio State, are Rebecca Pearson, Richard Robinson, Luanne Hall-Stoodley, Jacob Yount, Sonal Pannu and Rama Mallampalli.

The research was supported by a fund provided by an anonymous donor to Ohio State’s College of Veterinary Medicine, grants from the National Institutes of Health, a Glenn Barber Fellowship and an Ohio State COVID-19 Seed Grant.