Biology subjects

Banyard, A.

Publications and source records attributed to Banyard, A..

5 recordsLinked to original sources

bioRxiv2026

In situ programming of intratumoural stem cell-like memory CD8+ T cells enables durable antitumour immunity in immunosuppressive tumours

Tumours with an immunosuppressive microenvironment often respond poorly to radiotherapy (RT) and immune checkpoint inhibitors, and durable clinical responses remain rare. These failures are not solely explained by insufficient immune activation, but instead reflect an inability of current therapies to establish CD8 T-cell infiltration and states required for long-term tumour control. Although stem cell-like memory CD8 T cells (Tscm) are critical for durable antitumour immunity, they are typically rare within solid tumours, and strategies to selectively augment the expansion of intratumoural Tscm within immunosuppressive tumours are lacking. Here, we show that intratumoural immune modulation via pattern recognition receptor (PRR) activation, using a double-stranded RNA-based agonist (BO-112), promotes Tscm expansion and, in combination with radiation, enhances antitumour immunity in poorly immunogenic lung and bladder cancer models. PRR activation enabled selective in situ expansion of intratumoural Tscm, rather than global CD8 T-cell activation. Therapeutic efficacy required pre-existing intratumoural CD8 T cells and was associated with qualitative reprogramming of the immune landscape, including depletion of exhausted CD8 T cells and regulatory T cells. Transcriptomic analyses showed that PRR activation preferentially enriched programmes associated with Tscm formation and self-renewal, whereas durable effector differentiation emerged only following combined RT-PRR agonist treatment, consistent with antigen-driven execution of the expanded progenitor pool. A Tscm-associated gene signature derived from these preclinical datasets correlated with improved survival across independent human lung and bladder cancer cohorts. Together, our data identify intratumoural Tscm as a therapeutically targetable immune-control axis and establish in situ programming of Tscm as a route to durable antitumour immunity.

cancer biology↗

bioRxiv2023

Serological evidence of high pathogenicity virus infection in Eidolon helvum fruit bats in Nigeria

The Eidolon helvum fruit bat is the most widely distributed fruit bat in Africa and is known to be a reservoir for several pathogenic viruses that can cause disease in humans. To assess the risk of zoonotic spillover, we conducted a serological survey of 304 serum samples from E. helvum bats that were captured for human consumption in Makurdi, Nigeria. Using pseudotyped viruses, we screened the samples for neutralising antibodies against viruses from the Coronaviridae, Filoviridae, Orthomyxoviridae and Paramyxoviridae families. We report the presence of neutralising antibodies against henipavirus lineage GH-M74a virus (odds ratio 6.23; p<0.001), Nipah virus (odds ratio 4.04; p=0.00031), bat influenza H17N10 virus (odds ratio 7.25; p<0.001) and no significant association with Ebola virus (odds ratio 0.56; p=0.375) in the bat cohort. The data suggest a potential risk of zoonotic spillover including the possible circulation of highly pathogenic viruses in E. helvum populations. These findings highlight the importance of maintaining sero-surveillance of E. helvum to monitor changes in virus prevalence and distribution over time and across different geographic locations. Article summary lineThe detection of neutralizing antibodies against henipavirus GH-M74a virus, Nipah virus, and H17N10 virus in Eidolon helvum bat sera from Nigeria using pseudotyped viruses suggests a potential risk of zoonotic spillover.

microbiology↗

bioRxiv2020

Large extracellular vesicles can be characterised by multiplex labelling using imaging flow cytometry

Extracellular vesicles (EVs) are heterogeneous in size (30nm-10{micro}m), content (lipid, RNA, DNA, protein) and potential function(s). Many isolation techniques routinely discard the large EVs at the early stages of small EV or exosome isolation protocols. We describe here a standardised method to isolate large EVs and examine EV marker expression and diameter using imaging flow cytometry. MethodsWe describe step-wise isolation and characterisation of a subset of large EVs from the medulloblastoma cell line UW228-2 assessed by fluorescent light microscopy, transmission electron microscopy (TEM) and TRPS. Viability of parent cells was assessed by Annexin V exposure by flow cytometry. Imaging flow cytometry (Imagestream Mark II) identified EVs by direct fluorescent membrane labelling with Cell Mask Orange (CMO) in conjunction with EV markers. A stringent gating algorithm based on side scatter and fluorescence intensity was applied and expression of EV markers CD63, CD9 and LAMP 1 assessed. ResultsUW228-2 cells prolifically release EVs of up to 6 {micro}m. We show that the Imagestream Mark II imaging flow cytometer allows robust and reproducible analysis of large EVs, including assessment of diameter. We also demonstrate a correlation between increasing EV size and co-expression of markers screened. ConclusionsWe have developed a labelling and stringent gating strategy which is able to explore EV marker expression (CD63, CD9 and LAMP1) on individual EVs within a widely heterogeneous population. Taken together data presented here strongly support the value of exploring large EVs in clinical samples for potential biomarkers, useful in diagnostic screening and disease monitoring.

cell biology↗

bioRxiv2018

A simian-adenovirus-vectored rabies vaccine suitable for thermostabilisation and clinical development for low-cost single-dose pre-exposure prophylaxis

BackgroundEstimates of current global rabies mortality range from 26,000 to 59,000 deaths per annum. Although pre-exposure prophylaxis using inactivated rabies virus vaccines (IRVs) is effective, it requires two to three doses and is regarded as being too expensive and impractical for inclusion in routine childhood immunization programmes.\n\nMethodology/ Principal FindingsHere we report the development of a simian-adenovirus-vectored rabies vaccine intended to enable cost-effective population-wide pre-exposure prophylaxis against rabies. ChAdOx2 RabG uses the chimpanzee adenovirus serotype 68 (AdC68) backbone previously shown to achieve pre-exposure protection against rabies in non-human primates. ChAdOx2 differs from AdC68 in that it contains the human adenovirus serotype 5 (AdHu5) E4 orf6/7 region in place of the AdC68 equivalents, enhancing ease of manufacturing in cell lines which provide AdHu5 E1 proteins in trans.\n\nWe show that immunogenicity of ChAdOx2 RabG in mice is comparable to that of AdC68 RabG and other adenovirus serotypes expressing rabies virus glycoprotein. High titers of rabies virus neutralizing antibody (VNA) are elicited after a single dose. The relationship between levels of VNA activity and rabies glycoprotein monomer-binding antibody differs after immunization with adenovirus-vectored vaccines and IRV vaccines, suggesting routes to further enhancement of the efficacy of the adenovirus-vectored candidates. We also demonstrate that ChAdOx2 RabG can be thermostabilised using a low-cost method suitable for clinical bio-manufacture and ambient-temperature distribution in tropical climates. Finally, we show that a dose-sparing effect can be achieved by formulating ChAdOx2 RabG with a simple chemical adjuvant. This approach could lower the cost of ChAdOx2 RabG and other adenovirus-vectored vaccines.\n\nConclusions/ SignificanceChAdOx2 RabG may prove to be a useful tool to reduce the human rabies death toll. We have secured funding for Good Manufacturing Practice-compliant bio-manufacture and Phase I clinical trial of this candidate.\n\nAuthor summaryRabies was, after smallpox, the second human disease for which an efficacious vaccine was developed, by Pasteur in 1885. Although it is eminently preventable, with highly efficacious vaccines available for both humans and animals, it still causes considerable mortality in low and middle-income countries. It is a particular problem in areas with the weakest healthcare and veterinary infrastructure, where achieving prompt post-exposure vaccination or high-coverage dog vaccination are challenging.\n\nHere, we report the development of a new candidate rabies vaccine, designed to enable low-cost single-dose pre-exposure human rabies prophylaxis in such settings. ChAdOx2 RabG is based upon a simian adenovirus-vectored candidate previously shown to achieve protection after a single dose in non-human primates, now modified to allow clinical-grade bio-manufacture. We show that it induces a potent immune response in mice, that this response can be further enhanced by clinically-relevant adjuvant, and that we can stabilise it such that it can withstand temperatures of up to 45 {degrees}C for a month. We will be performing a clinical trial of this candidate in the near future.

immunology↗

bioRxiv2017

Generation of Arctic-like Rabies Viruses Containing Chimeric Glycoproteins Enables Serological Potency Studies.

Rabies viruses have the highest case fatality rate of any known virus and are responsible for an estimated 60,000 deaths each year. This is despite the fact that there are highly efficacious vaccines and postexposure prophylaxis available. However, while it is assumed these biologics provide protection against all rabies virus isolates, there are certain subdivisions of RABV lineages, such as within the Arctic-like RABV (AL rabies virus lineage, where data is limited and thus the potency of existing biologics has not been thoroughly assessed. By fusing the Arctic-like rabies virus envelope glycoprotein ecto- and transmembrane domains with the vesicular stomatitis virus cytoplasmic domain, a high titre (7.7 x 105 - 6.1 x 106 RLU/ml) pseudotyped virus was generated that was subsequently used in a pseudotyped virus neutralisation assay. These results showed that Arctic-like rabies viruses are neutralised to human, canine and feline vaccines and human post-exposure prophylaxis and this was not influenced by the swapping of the cytoplasmic domains (CVS-11 vs CVS-11etmVSVc; r = 0.99, p < 0.0001). This study supports the concept that rabies virus vaccines and newly identified mAbs are able to neutralise rabies virus variants that cluster in a monophyletic clade, referred to as phylogroup I lyssaviruses.

microbiology↗