Microwave Radiation as a Method for Killing Covid and Other Viruses?

 You can find the article here, and you can find more from the Air Force Research Lab here. 

Introduction

During the Sars-Cov-E-2 pandemic (COVID-19), numerous methods were attempted to reduce viral transmission, including temperature variation in both hot and cold weather (Who), alcohol (BBC),  and bleach (Who). Despite many of these ideas, which were the result of misunderstood science, falling short of decreasing viral transmission and destroying the virus, several methods which showed previous interesting results were brought back into the limelight to be reanalyzed. Of these, the use of non-thermal microwave radiation to kill COVID-19 was explored, despite previous results indicating microwave radiation performing poorly against viral infection (Epstein and Cook 1951). In recent years, evidence has shown that non-thermal radiation inactivation may host a proposed mechanism involving the electromagnetic coupling of microwave radiation to mechanical dipole resonances, meaning microwaves can cause mechanical stress if at the right frequency (Lui et al 2009). This effect, known as the structure-resonant energy transfer (SRET), has been seen in previous explorations which have covered SRET and thermal effects on pathogen inactivation (Xiao et al. 2022).  Researchers hypothesized that high-powered microwaves may cause more mechanical damage to viruses, but studies indicated that inactivation of viruses from microwaves was less than 75% (Cantu et al. 2023). In this study, four frequencies of microwave which were noted as being “of interest” for inactivation of Bovine Coronavirus (BCoV) were explored for BCoV in aerosols. Previous studies by this group (Hoff et al. 2023) showed roughly 74% inactivation of BCoV in aerosol when exposed to 5.6 GHz. In order to obtain a more comprehensive evaluation of microwave-virus interactions, this study hypothesized that exposing aerosolized BCoV to a lower frequency of microwave, namely 4.0 GHz would result in weaker inactivation compared to 5.6 GHz. 

Methods

Cohick et al. used a custom built experimental apparatus which allowed them to pump aerosolized BCoV down a tube which had consistent radiation. This set-up included aerosol being generated using a nebulizer. Once generated, the aerosol would flow down a polycarbonate flow tube into a biosampler.

Figure 1) Custom apparatus used to explore radiation effects on aerosolized viruses, including the collision nebulizer (left) and the biosampler (right) and the radiation input (center). 

Findings

 46 experiments were carried out (20 with RF exposure and 26 control/no-RF exposure).  Each experiment took 15 minutes to complete. After the virus was aerosolized and irradiated, virus survival rates were analyzed using a TCID50 assay, which is a method of determining the infectious virus titer by calculating the dilution required to produce a known cellular effect, such as cell death. Their results indicated there was no statistically significant reduction in BCoV survival rate while under 4.0 GHz radiation in comparison to BCoV under no radiation. There was also no statistically significant increase in the standard deviation when comparing the results under RF exposure and control. 

Figure 2) BCoV survival rate for irradiated samples and control samples. Median and interquartile ranges are depicted, alongside raw data.

Conclusion

Given previous results showing that BCoV had a strong reduction in survivability under higher frequencies but a lessening reduction in survivability under smaller frequencies (Cantu et al. 2023) this result is consistent.  Furthermore, this trend is noted as aligning with observations in other SRET work involving the neutralization of viruses. Cohick et al. plan to continue experiments on aerosolized BCoV under high frequencies and exposures of BCoV virus to shorter RF pulse lengths. They hope to create a more comprehensive understanding of how frequency-based radiation may affect the survivability of COVID-19.

Citations

Cohick, Z. W., Hoff, B. W., Revelli, D., Cox, J., Irshad, H., Snider, A., Arndt, A., Enderich, D. A., McCohaha, J. W., Schrock, J. A., Ibey, B. L., Thomas, R. J., Luginsland, J. W., Roach, W. P., & Shiffler, D. A. (2026). Evaluation of 4.0 ghz RF exposure effects on bioaerosols containing bovine coronavirus. Bioelectromagnetics, 47(1). https://doi.org/10.1002/bem.70040 

BBC. (2020, March 5). Coronavirus: Don’t use vodka to Sanitise Hands. BBC News. https://www.bbc.com/news/business-51763775 

WHO. (2022, January 19). Covid-19 mythbusters. World Health Organization. https://www.who.int/emergencies/diseases/novel-coronavirus-2019/advice-for-public/myth-busters 

Epstein, M. A., and H. F. Cook. 1951. “The Effects of Microwaves on the Rous No. 1 Fowl Sarcoma Virus.” British Journal of Cancer 5, no. 2: 244–251. https://doi.org/10.1038/bjc.1951.25.

Liu, F., B. Rittmann, S. Kuthari, and W. Zhang. 2023. “Viral Inactivation Using Microwave‐Enhanced Membrane Filtration.” Journal of Hazardous Materials 458: 131966. https://doi.org/10.1016/j. jhazmat.2023.131966.

Xiao, Y., L. Zhao, and R. Peng. 2022. “Effects of Electromagnetic Waves on Pathogenic Viruses and Relevant Mechanisms: A Review.” Virology Journal 19, no. 1: 161, 2022/10/12. https://doi.org/10.1186/s12985-022-01889-w. 

Cantu, J. C., J. W. Butterworth, K. S. Mylacraine, et al. 2023. “Evaluation of Inactivation of Bovine Coronavirus by Low‐Level Radiofrequency Irradiation.” Scientific Reports 13, no. 1: 9800. https://doi.org/10.1038/s41598-023-36887-7.

Hoff, B. W., Z. W. Cohick, B. S. Tilley, et al. 2023. “Observed Reductions in the Infectivity of Bioaerosols Containing Bovine Coronavirus Under Repetitively Pulsed RF Exposure.” IEEE Transactions on Biomedical Engineering 70, no. 2: 640–649. https://doi.org/10.1109/TBME.2022. 3199333.

ZFP36LR: The Latest Anti-HIV Inhibitor Makes its Debut

 You can find the original article here, and the First Hospital of China Medical University here.

Introduction

Human Immunodeficiency Virus, or HIV, is a well-known group of viruses that has eluded sterilizing treatment for over four decades. It can be transmitted sexually (usually via unprotected intercourse resulting in contact with bodily fluids), but also vertically (from mother to child) and via blood transfusion (1). HIV infection first presents, usually in the weeks following initial infection, in an acute (immediate) phase via flu-like disease, including fever, malaise (non-specific discomfort), and rash. This is followed by a long-term asymptomatic phase, where the patient usually experiences no disease for years following infection (2). This phase is usually the result of seroconversion, wherein the patient’s CD4+ T Cells create antibodies (small molecules designed to attach to and stop the activity of the outer components of viruses) capable of eliminating the virus. However, over the course of years, the virus, which persists in low numbers, gradually eliminates the patient’s CD4+ T cells and adapts to the antibodies by changing its outer protein shape (1). This causes a gradual increase in the amount of virus the patient has, and puts them at risk for opportunistic infection (when other viruses, bacteria, fungi, or parasites take advantage of the patient's weakened defenses to infect them) (1). Once the concentration of CD4+ T cells falls below a certain concentration (200 cells per milliliter of blood), the patient is considered to have Acquired Immunodeficiency Syndrome, or AIDS (2). Death typically results from the patient being unable to fight off other infections due to the elimination of their immune cells. 

Despite forty years of attempting to create a vaccine capable of reducing HIV’s burden amongst patients, no such treatment has (yet) materialized (3). Thus, the attention of the scientific enterprise has shifted toward attempting to fully understand how HIV disarms our immune cells and the ways our immune system attempts to fight back. Such studies have resulted in the creation of antivirals, small molecules that prevent the virus from replicating. Combinations of these antivirals have served as effective and life-changing treatments (1). Whereas HIV infection was once a death sentence, it has become a manageable condition. 

The first line of defense our immune system has against any viral infection is its interferon (IFN) response (1). In the case of HIV-1, the interferon response is mediated by a specific class of molecules called type I interferons (IFN-Is). These molecules, upon the detection of a virus, force our cells to undergo a huge amount of internal changes, which place them into a sort of “antiviral mode”. These changes are mediated by Interferon-stimulated genes (ISGs), which change the concentration of our cells’ internal components in a way that mitigates viral spread or replication (1). The sheer number of ISGs and their effects on our cells and immune system are so numerous that science has yet to exhaustively identify them. This paper, from the State Key Laboratory of the First Hospital of China Medical University, uncovers the novel role of an ISG-induced inhibitor: ZFP36L2. 

A Novel ISG-induced inhibitor: ZFP36L2 Mitigates HIV-1 Infection of Cells

Figure 1A, D: These results show that ZFP36L2 can prevent HIV-1 viruses from forming in immune cells.

The authors first tested the effect of adding ZFP36L2 to regular cells (A) or immune cells (D) to see if it had any effect on HIV-1 virus formation. They measured p24, a component of HIV-1’s outer shell, to measure the amount of virus being made. As can be seen above, adding ZFP36L2 causes a massive decrease in p24 concentration, indicating that it can potentially stop HIV-1 replication in T Cells. HIV-1 typically causes death because its long-term replication in CD4+ T cells leads to their elimination. This leaves the patient extremely vulnerable to parasites, fungi, bacteria, and other viruses that are normally dealt with by these cells. Thus, if ZFP36L2 can stop HIV-1 from replicating in T Cells, it may be possible to use it to prevent these opportunistic infections and other damage caused by the virus. In subsequent tests, the authors found that ZFP36L2 specifically targets “primate lentiviruses” (a class of viruses that infect humans and primates, of which HIV-1 is one member), which means that it may be possible to use it to fight HIV-1 without triggering the entire immune system (which can also be dangerous). Finally, the authors find through yet more experiments that ZFP36L2 is indeed an interferon-stimulated gene-induced inhibitor, becoming activated by the presence of IFNβ.

ZFP36L2: How does it work?

After the authors established that ZFP36LR is an ISG-induced inhibitor capable of reducing HIV replication in immune cells, they sought to understand more fully, and to the molecular level, how the protein worked. At this scale (the scale of proteins), activity is mainly determined by shape. You can think of proteins as highly complex puzzle pieces capable of interacting with other proteins, which can change what they can do. Thus, the authors ran further experiments to figure out which proteins ZFP36L2 interacted with, as doing so could provide insights on how to use designed molecules to activate ZFP36L2, and thereby fight HIV infections. In their first set of experiments, the authors find that when ZFP36L2 is given to cells, there are remarkable changes to the movement of genetic materials (Fig. 2G). When HIV-1 is ready to build new viruses, it needs to deliver its genetic code from the nucleus, where they’re created, to the cytoplasm, where they are translated into proteins that form the viruses. However, when ZFP36L2 is present, this material remains in the nucleus, except for fully spliced transcripts. In general, before human cells can “translate” genetic material into proteins, they must first be cut and fashioned into specific configurations. These fully configured materials pass through unaffected, but partially configured materials (including HIV-1’s genetic code) remain stuck in the nucleus. Thus, it seems that ZFP36LR prevents HIV-1 from passing down its genetic information, which is needed to make successful viruses. Notably, all the arrested materials contain a Rev Response Element, or RRE. This is, essentially, a genetic “pattern” that the HIV-made protein Rev can interact with (that is, it has a specific shape). These points lead the authors toward exploring Rev’s interactions with ZFP36L2.

Fig 3B,D: These graphs show the level of genetic material being made without and with ZFP36L2, without (B) and with (D) the aforementioned RRE.

The above results show that, when RRE’s are present (such as on the HIV’s genetic code) and ZFP36L2 is present, the amount of genetic material being created massively decreases. This indicates that ZFP36LR uses Rev to interact with RREs and prevents them from leaving the nucleus, thereby preventing HIV-1 from acquiring copies of its genetic code. Later experiments confirmed that ZFP36LR directly interacts with Rev.

 

ZFP36LR: Can it Really Treat Patients in the Real World?

Having found whether and how ZFP36LR can mitigate HIV-1 infection of immune cells is astonishing. However, these experiments, like many, were performed using highly modified cells in conditions wholly alien to those inside a patient. Thus, while a relationship between ZFP36LR and HIV infection can be established, it is still difficult to definitively say whether activation of ZFP36LR could be used as a future antiviral against HIV-1 in patients. At the same time, doing such experiments on living humans comes with major ethical and moral concerns, as these are highly experimental and thereby risky techniques. Thus, the authors decided to use an ex vivo model. When scientists perform experiments like these, they tend to use immortalized cells (cells that have been modified or otherwise mutated to replicate indefinitely) in highly artificial environments. These are known as in vitro experiments. Performing experiments on a patient would be in vivo. Ex vivo, then, occupies a sort of middle ground. In this case, the authors extracted CD4+ T cells from HIV patients and performed experiments on them in an artificial environment. These cells will be nearly identical to those in patients, so the results from such experiments will be more applicable in clinical settings. At the same time, these experiments come at no risk to any would-be test subjects. Thus, with an ex vivo model, the authors tested to see the effect of adding IFNβ to HIV-1 replication and ZFP36LR concentrations.

Fig. 6I, J: These graphs show the concentrations of virus (I) and genetic material encoding ZFP36LR (J) with and without IFNβ.

As can be seen above, when IFNβ is added to the patients’ cells, the amount of virus massively decreases, while the concentration of genetic material creating ZFP36LR massively increases. When ZFP36LR creation is blocked, this decrease in HIV-1 concentration disappears, indicating that activating IFNβ in patient cells is capable of activating ZFP36LR and mitigating HIV-1 replication. 

Conclusion

The set of experiments the authors conducted demonstrates that ZFP36LR is a protein created by an ISG, which is activated by IFNβ, and interacts with HIV-1’s genetic code to prevent it from replicating. More excitingly, the ex vivo experiments indicate that IFNβ activation can successfully mitigate HIV-1 infection of immune cells within patients through this mechanism, which has massive implications for antiviral design. IFNβ is unfortunately a pleiotropic protein (a protein with many roles and functions), so it’s not feasible to simply activate it in HIV-1 patients. Doing so would have any number of side effects, given its central role in the wider antiviral responses of our cells. ZFP36LR, by contrast, seems to be specific to targeting HIV-1, so future antivirals could come in the form of IFNβ analogues (molecules that mimic the shape of IFNβ, but that only activate ZFP36LR). As the battle against HIV-1 wages on, studies like these uncover ever more targets against HIV-1, expanding the options for treatment for HIV-1-infected patients. Although a sterilizing treatment for HIV-1 continues to elude science, discoveries such as these continue to improve the outcomes of millions living with HIV. 

References

Gilroy, S.A., (2025, January 3). HIV Infection and AIDS Clinical Manifestation. Medscape, 

https://emedicine.medscape.com/article/211316-clinical.  

(No Authors), (2025, January 14). About HIV. Center for Disease Control, 

https://www.cdc.gov/hiv/about/index.html. 

Shim, I., Rogowski, L., & Venketaraman, V. (2025). Progress and Recent Developments in HIV 

Vaccine Research. Vaccines, 13(7), 690. https://doi.org/10.3390/vaccines13070690.  

Pang, H., Cui, H., Yin, X. et al., (2026). ZFP36L2 is an interferon β -induced inhibitor that 

restricts the nuclear export of HIV-1 transcripts. Nature Communications,  https://doi.org/10.1038/s41467-026-71474-0.

Examining the Environment to Identify Viruses in Livestock

 With the increased  reliance on livestock and animal products in today’s world, humans and animals are in closer proximity than ever. This closeness however, also makes this connection susceptible to be corrupted by the spread of viral disease. This spread from animals to humans is called zoonotic infection and is how many of the most serious viral outbreaks began such as influenza and coronavirus. Livestock are particularly vulnerable to the spreading of these viruses as they are often clustered close together, promoting viral transmission that can eventually make its way to human hosts. Resources for monitoring viruses within livestock populations have, however, been a challenge as traditional monitoring has had to be done directly on the animals which can pose risks to both the animals and the researchers. Thus, scientists are looking to develop new ways to surveil virus transmission in non-invasive ways.

One way researchers have looked to solve this problem is that rather than testing the animals themselves, they could look at samples of aggregated materials from many animals such as manure, air samples, and wastewater to track viruses within animal populations. Methods of monitoring pathogens from aggregated samples have already proven effective in some cases and researchers wanted to further test this theory to see if aggregate samples could appropriately serve as reliable indicators of virus transmission in  populations of cattle and pigs.

The researchers began by setting up sample collection sites at cow and pig farms in Barcelona as well as at several slaughterhouses. They collected 105 samples across eleven months between slurry, manure, wastewater, and air samples from the locations. The samples were separated into different pools based on whether they were from pigs or cows and by the season they were collected. Viruses were then extracted from each sample by slowly removing organic and biological particles from the sample until viral DNA could be isolated.

In the swine samples, the researchers found evidence of 56 total viral species and saw that the viruses seemed to have seasonal variation with each virus seeming to have a preferred peak season of high abundance that matches with known viral seasonality indicating a success of using aggregated samples to test for viruses.

The researchers then compared their findings with the samples taken at cattle farms and from cattle wastewater samples. They found that since cows are housed at a lower density than pigs, there was a sharp decrease in pathogen detection since viruses could not be passed as easily when the animals are farther apart. The researchers did however notice the same pattern of seasonality for the 63 viral species that appeared in their screens, many of which appeared in a specific sample type alone showing the best ways to monitor those specific viruses. 

Based on their results, the researchers conclude that an aggregated sample approach is an effective way to monitor animal viruses in livestock populations. Based on their results and their alignment with previous direct studies on viral presence in livestock populations, the authors believe that aggregate sampling could prove to be of significant use, especially in the prevention of zoonotic infections that could infect humans. While this method can make it difficult to determine which individuals in a population are infected due to the collection methods, the overall results from this study indicate that there are some new approaches coming to virus monitoring and the prevention of virus spread.

In order to be even more effective with these new techniques for virus monitoring, we could complement them through making advances in viral detection technology that might allow us to track viruses and their mutations so we can be even more prepared to face viral threats as they develop before they infect the larger population. We could also look into applying this technique to non-livestock animals which would help to prevent the spread of infection globally. This paper demonstrates a major shift coming for viral monitoring and detection which could ultimately lead help us to be safer and more prepared for emerging viral infections

Literature Cited:

Rusiñol, M., Martínez-Puchol, S., Ribeiro, D., Verdaguer, J., Torrejón-Llorens, O., Itarte, M., Estarlich-Landajo, I., Mejías-Molina, C., Juliachs-Torroella, G., Girones, R., Ramírez, G. A., Baliellas, J., Bofill-Mas, S., & Fernández-Cassi, X. (2026). Livestock aggregated samples for monitoring viruses infecting animals and potentially zoonotic viral pathogens. One health (Amsterdam, Netherlands), 22, 101340. https://doi.org/10.1016/j.onehlt.2026.101340

A Rabies-Based Vaccine? The Emerging Vaccination Landscape.

 Over the last couple of years, many scientists and epidemiologists have been watching the spread of a new virus, the H5N1 avian influenza. While initially developed in birds, there has just been a recent development where it has now begun to spread within populations of cattle, and even humans have now been infected. This new flu is extremely worrying as though the infected humans only seem to have mild symptoms, previous H5N1 epidemics have had severe mortality rates. Thus, scientists are currently looking for some possible vaccinations that work for humans and animals, should this virus become a major public threat.

One possible vaccine avenue that scientists are interested in studying is through the use of dual rabies virus vaccines or RABV that have been known to protect against other viral pathogens. To test this theory, a lab at Thomas Jefferson University developed a RABV-based H5 dual vaccine or RABV-H5 that has antigens which would protect against both the avian influenza as well as rabies virus. 

In order to develop this vaccine, the researchers inserted a synthesized version of the avian flu into the genetic sequence of rabies virus. The proper functioning of both parts of the new vaccine sequence were then analyzed and confirmed that the vaccine sequence had effective functioning of both viruses and that the influenza sequence had successfully integrated into the rabies virus sequence.

Figure 1A.

Now that the vaccine had been made, the researchers wanted to determine how effective this vaccine was. The researchers tested the vaccine on mice using different methods to vaccinate them. They used both an inactivated, or killed, form of the RABV-H5 as the vaccine as well as a live version of RABV-H5 and vaccinated the mice either with an injection in the muscle or by applying droplets of vaccine to the nose. 

Figure 1E.

After vaccination, the researchers looked for antibodies to the H5 which would serve as a sign that the vaccine worked and led to immunity of the virus. They found that in all cases, the mice had antibodies to H5 and found that vaccinating with a muscle injection at both the primary and secondary vaccination steps, the mice had higher antibody levels which indicates that these mice would have the strongest viral immunity.

After the success of the initial vaccination test, and following the same vaccination procedure paired with non-vaccinated control mice, mice were given a lethal dose of a mouse adapted version of the avian influenza virus. The researchers found that mice given the vaccine showed fewer signs of illness including weight loss and less virus seen on their lungs and had a much higher survival rate, including a one hundred percent chance of survival when given the vaccine intramuscularly both times or intramuscularly the first time and then a live version of the RABV-H5 vaccine delivered intranasally as a booster. This result stands in stark contrast to the mice that were unvaccinated and succumbed to the virus within six days, and even the mice that were vaccinated in alternate ways that only had a forty to sixty percent survival rate.

Seeing this success, the researchers did the exact same experiment, but this time with the authentic version of the avian influenza. Similar results followed from this experiment as vaccinated mice had a high survival rate and saw limited effects from the virus while unvaccinated mice succumbed to the virus at six to seven days.

With the success of the vaccine against both the mouse adapted version of the virus and the authentic virus itself, the researchers also wanted to look at how the vaccine would perform against a similar virus strain that is currently in circulation.  The researchers found that the vaccine did seem to have some effect as it reduced weight loss in the vaccinated mice and led to high survival rates against a reduced form of the virus, but as the researchers expected, the vaccine had no effect on the lethal dose of the virus.

This paper illustrates the wide world of vaccines and the unlikely places that they can come from. The use of vaccine platforms such as RABV to build new vaccines in quick order so as to prevent disease spread should not be underestimated as these platforms most likely hold the key to preventing the spreading of emerging viruses and combatting viruses before they become a major human concern. The virus created and tested in this paper also offers a glimpse at emerging vaccine technology such as the ability to vaccinate for multiple viruses with a single vaccine and also shows the increasing need to develop vaccines with both humans and animals in mind to prevent a virus from spreading from animals to humans.

While this study shows a lot of exciting potential for vaccines against new and emerging infectious diseases, this study leaves open the future possibilities of this vaccine as the next steps would be evaluating its effectiveness in humans after the successes seen in mice. While there may be future viral threats that could begin a new epidemic, this emerging vaccine technology could help us to prevent that from happening.

Literature Cited:

Paran, N., Wirblich, C., Albrecht, R., Zabihi-Diba, L., Tarquinio, A., Kurup, D., Solomides, C. C., García-Sastre, A., & Schnell, M. J. (2026). Immunogenicity and efficacy of a rabies-based vaccine against highly pathogenic influenza H5N1 virus. Emerging microbes & infections, 15(1), 2620221. https://doi.org/10.1080/22221751.2026.2620221

Combatting Against Cancer Using Virus: Oncolytic Virus M1 Reinvigorates T-cell immunity against glioblastoma

Glioblastoma multiforme (GBM), usually arising from glial precursor cells or neural stem cells, is the most common and lethal primary brain cancer in adults. It has very limited treatment options and a median life expectancy less than 2 years after diagnosis. Systemic and local immunosuppression induced by GBMs largely contributes to malignancy aggressiveness and resistance to multiple immunotherapies, such as immune checkpoint blockade (ICB) therapy, which is powerful for primary care of other types of solid tumors. 

Oncolytic virotherapy has attracted growing awareness as a potential transformative cancer therapy in recent years. It involves delivery of a genetically modified version of virus into the host and it would selectively destroy tumor cells while leaving neighboring healthy cells undamaged, overcoming a common limitation of chemotherapy. This targeted destruction is either achieved through direct oncolysis ("onco-" meaning cancer-related, "-lysis" meaning burst) or immune-mediated attack. Oncolytic therapy using alphacirus M1(OVM), which was originally isolated from Culex mosquitoes, recently emerged as a potential glioma therapeutic approach. The virus is delivered through intravenous administration and the main advantages are OVM's ability to selectively replicate in tumor cells overexpressing matrix remodeling-associated 8 (the marker allowing them to recognize and attach) and efficiency in penetrating through the blood brain barrier, which is a dense layer of blood vessels and cells that protect our brain from harmful substances but makes drug delivery difficult at the same time. Even though we might be worried that using virus as medium can be dangerous by itself at the first glance, the toxic genes are being taken out in advance and it's previously shown to be very safe: GBM patients showed high tolerance towards OVM in previous clinical trials and repeated intravenous administration of OVM is non-pathogenic for nonhuman primates (Zhang et al., 2016).

Despite of its huge therapeutic potential, the knowledge regarding how OVM initiates adaptive immunity against gliomas and how the interactions between OVM and our immune system would influence the therapeutic effects of ICB is still lacking. Hence the researchers are seeking to address these two knowledge gaps.

First, they confirmed the efficacy of OVM in reducing glioma progression. The immunocompromised mice were injected intracranially with GBM cell lines (GL261-Luciferase, GL261 and CT2A) to induce a glioma in the brain on day 0 and followed by daily tail intravenous injection of OVM or vehicle from day 5 to day 9. Vehicle includes the buffer solutions that researchers use to deliver OVM but containing no OVM, acting as the control condition. GL261-Luciferase is a cell line where it's infected with lentivirus whose genome was engineered to contain the luciferase gene so that wherever the tumor develops, it would glow. A stronger luminescence intensity (red) indicates the progression of glioma while bluish color indicates little tumor growth. It was found that OVM significantly inhibited the growth of glioma and prolonged the survival of glioma-bearing mice. 

Moving on to investigate the immune response, the researchers found that OVM successfully induced immunogenic cell death and reverse the GBM-induced immunosuppression both locally and systemically. Immunogenic cell death is characterized by the release of damage-associated molecular patterns (DAMPs) by tumor cells, which are molecules that normally stay inside the cell and becomes 'danger signals' recognized by immune cells when being released outside, driving adaptive-immunity elimination in tumor microenvironment. They examined the release of DAMPs, revealing significant CRT (externalized calreticulin, a multifunctional protein) exposure and elevated extracellular ATP (adenosine triphosphate, energy source for our cells) in the supernatant collected from glioma cell lines with OVM treatment. This indicates the occurrence of immunogenic cell death induced by OVM. 

Additionally, the number of CD4+ and CD8+ T cells increased significantly in peripheral blood of glioma-bearing mice following intravenous OVM treatment. Spleen atrophy and loss of splenic T-cell population were also recovered. Similarly, in the tumor local environment, they also found rapid increase in tumor-specific CD8+ T cells in the spleen. The measurement is achieved by orthotopical implantation of GL261 glioma cells expressing OVA (full-length ovalbumin, processed into SIINFEKL) and H-2Kb-SIINFEKL tetramer that binds to T cell receptors where H-2Kb is the mouse version of MHC class I protein. The recognition of glioma-derived-antigen-MHC complex by T cells allows the accurate tracking of tumor-specific T cells (Hsu et al., 2025). Hence, with OVM infection, both local and systemic immunity are boosted and recovered.

  

Before diving deeper into the mechanism, the researchers digressed a bit and addressed a main concern about the efficacy of OVM: Spleen is the major body 'filter' consisting of large number of immune cells that's found previously able to eliminate delivered oncolytic viruses and OVM is often 'trapped' inside the spleen of various host animals. Therefore, the researchers removed the spleen (splenectomy) from mice to find out the role of spleen in OVM treatment. To their surprise, opposite to the greater reduction in glioma progression that they have expected, the antitumor activity of OVM treatment was completely abrogated after removal. Splenectomy-OVM mice almost have the same low survival probability as vehicle control and no expansion of T-cell population in the peripheral blood and tumor microenvironment.

Consequently, the researchers tried to find out the specific cell population in the spleen that's mediating the reversal of immunosuppression and antitumor activity. They first employed a broad RNA sequencing and computational screening and found that B cells have the highest abundance and showed strongest predicted interactions with T cells after OVM treatment. To move beyond correlation, a series of validation experiment were conducted: In vitro co-culture experiments demonstrated that B cells from OVM-treated mice significantly enhanced activation of T cells but not other splenic immune populations and this activation is dose-dependent. Further, after blocking the B cells with anti-CD19 antibody, OVM administration failed to prolong the survival of glioma-bearing mice and increase the infiltration of T cell populations in the blood and tumor microenvironment. 

To elucidate the mechanism by which splenic B cells reverse immunosuppression, the researchers compared the interactions of B cells with CD8+ T cells in the OVM-treated group and the vehicle control group. They found that OVM treatment leads to much more frequent physical contacts between B and T cells, visualized through live cell imaging.

To determine the functional significance of this cell-to-cell contact, they created two conditions where the cells can directly interact with each other and where they are forced to be separated from each other by a membrane (transwell chamber). CD8+ T cell proliferation is measured by CFSE dilution (carboxyfluorescein succinimidyl ester,  a fluorescence dye that's split between new-born cells for every division) and CD8+ T cell activation is measured by GZMB+ (granzyme B, the 'weapon' T cells produce to kill target cancer cells). Using these two tracking tools, they discovered that T cell proliferation and activation is induced only when B cells and T cells that can freely contact with each other but not in transwell-separated groups. These results together suggests that the direct contact between B and T cells is essential for T-cell expansion and activation required for OVM-mediated immune enhancement.

In order to validate this finding in vivo, researchers targeted the Major Histocompatibility Complex class I (MHC-I), a critical component of the structural interface between B and T cells. B cells communicate and 'educate' T cells by presenting a fragment of tumor on MHC-I to activate them, which process is known as antigen presentation. Blocking MHC-I with a specific antibody completely abolished B-cell-induced CD8+ T cell proliferation and activation. Collectively, these results confirm that the direct physical interface is crucial for immunity restoration by OVM treatment. Later the researchers identified the specific subset of B cells that's responsible for this interaction to be Bst2+ B cells by computational screening and in-vivo evidence: Bst+B cells significantly prolonged the survival of B-cell-deficient, GL261-OVA-bearing mice after OVM stimulation while Bst2-B cells and B cells from vehicle control group did not.

The interface between B and T cells involves B cells 'presenting' a peptide derived from cancer cells or other sick cells on MHC-I protein, which is then recognized by T cell receptors (TCR). 

Finally, to bridge the above mechanistic findings with clinical application, the researchers tested if OVM treatment can help overcome immunosuppression that previously has prevent immune checkpoint blockade (ICB) therapy from functioning in glioblastoma patients. ICB therapy works by removing the "brake" on the T cells. Our bodies have these natural immune checkpoint proteins that's naturally used to prevent excessive inflammation from happening that would otherwise harm other healthy cells. But at the same time, this also grant tumor cells a tool to escape from immune responses. ICB drugs block these inhibitory signals and boost the working capacity of our immune system. 

PD-L1 and PD-1 are the immune checkpoint proteins that 'put a brake' on the T cell when bound. If either of them are blocked, which is what the researchers and ICB drugs do, T cells are free to attack the tumor cells. 

To test the synergy between ICB and OVM therapy, immunocompromised mice was injected intracranially on day 0 and from day 5 to day 9, the mice is divided into different groups and treated daily intravenously with different combinations of immune checkpoint protein antibodies, including isotype antibody as control, PD-1 and PD-L1 antibodies, with or without OVM. The researchers revealed that, comparing with other 5 combinations, OVM treatment and PD-1 inhibitor together yielded the greatest recruitment of CD8+ T cells into tumor microenvironment, suppression of intracranial glioma growth as well as restoration of splenic size and weight. Systematically, massive expansions of effector (activated) CD8+ and CD4+ T cells in peripheral bloodstream are also observed. Therefore, OVM synergizes with PD-1 inhibitors for glioma treatment by simultaneously reshaping systemic immunity and the local immune environment in the brain.

isotype= control isotype antibody; PD-1 and PD-L1 Ab= PD-1 and PD-L1 antibodies (ICB therapy); Vehicle= control for OVM treatment; OVM= oncolytic viral therapy. 

This study provides profound mechanistic details for how emerging OVM treatment reverses the immunosuppressive environment of GBM, which is incurable and has grim prognosis. Most significantly, it establishes OVM as a transformative catalyst for ICB therapy, which is historically ineffective for GBM. The findings also illuminate a remarkable capability of OVM bypassing the 'toxic', immunosuppressed tumor microenvironment in the brain, and re-directing the site of immune cell communication to the 'clean sanctuary', the spleen. Within this protected environment, the recovery of B- and T-cell physical interface structures allows for robust antigen presentation, subsequent T cell activation, and eventually elimination of the tumor cells. The study has a strong advantage in its comprehensive validation of each step. For instance, when assessing the efficacy of the therapies, they would use several measures like the size of the spleen, tumor growth and immune cells activation level. Also, when confirming the determining role of splenic B cells, they combined both computational tools and biological checks, including knockout and correlation experiments. Moving forward, these discoveries open a new frontier for translational medicine. While mouse models provide an essential foundation, future research should evaluate OVM in non-human primates and eventually human clinical trials to characterize its safety profile in more complex in vivo environment. Long-term studies are also preferable because so far only the decrease in size of tumor is. reported but future relapse is still possible, especially given the ability of tumors to develop resistance mutations. Furthermore, the mechanism behind OVM-directed B-cell migration to the spleen and T-cell migration to the brain can be elucidated to identify potential signaling molecule targets.

Citations:

For images:

https://www.researchgate.net/figure/The-origin-of-glioblastoma-During-normal-embryonic-development-and-in-the-adult-brain_fig2_371833392

https://vyriad.com/science/oncolytic-virus-platforms/
https://www.cancerresearch.org/immunotherapy-by-treatment-types/oncolytic-virus-therapy
https://www.nature.com/articles/s41592-020-01031-0
https://www.cancer.gov/about-cancer/treatment/types/immunotherapy/checkpoint-inhibitors

https://rcastoragev2.blob.core.windows.net/13974d8439d3384abb2302e11f2da069/PMC9563749.pdf

For academic literature:
the primary article:
‌Han, Yu, et al. “Oncolytic Virus M1 Reinvigorates CD8+ T-Cell Immunity against Glioblastoma through B-Cell-Dependent Antigen Cross-Presentation in the Spleen.” Cellular & Molecular Immunology, 4 Mar. 2026, www.nature.com/articles/s41423-026-01396-w/figures/8, https://doi.org/10.1038/s41423-026-01396-w. Accessed 26 Mar. 2026.

Others:

‌National Cancer Institute. “Https://Www.cancer.gov/Publications/Dictionaries/Cancer-Terms/Def/Blood-Brain-Barrier.” Www.cancer.gov, 2 Feb. 2011, www.cancer.gov/publications/dictionaries/cancer-terms/def/blood-brain-barrier.

‌Hsu, Chung-Yao, et al. “Polymerised Superparamagnetic Antigen Presenting Cell Lymphocyte Capture for Enriching Tumour Reactive T-Cells and Neoantigen Identification.” Nature Communications, vol. 16, no. 1, 2 June 2025, www.nature.com/articles/s41467-025-60321-3, https://doi.org/10.1038/s41467-025-60321-3. Accessed 26 Mar. 2026.

“Up Close: How Immune Checkpoint Inhibitors Revolutionize Cancer Treatment | What’s up at Upstate | SUNY Upstate.” Www.upstate.edu, www.upstate.edu/whatsup/2023/100523-up-close-how-immune.php.

Zhang, Haipeng, et al. Naturally Existing Oncolytic Virus M1 Is Nonpathogenic for the Nonhuman Primates after Multiple Rounds of Repeated Intravenous Injections. Vol. 27, no. 9, 1 Sept. 2016, pp. 700–711, https://doi.org/10.1089/hum.2016.038. Accessed 1 Aug. 2023.

‌“Defined Immune Response Tracking in Mice: OVA MHC Tetramers – Caltag Medsystems.” Caltagmedsystems.co.uk, Mar. 2024, www.caltagmedsystems.co.uk/information/defined-immune-response-tracking-in-mice-ova-mhc-tetramers/. Accessed 29 Apr. 2026.

‌“Oncolytic Virus Therapy and Its Side Effects.” Cancer.org, 2023, www.cancer.org/cancer/managing-cancer/treatment-types/immunotherapy/oncolytic-virus-therapy.html.

‌

‌