Abstract
Nipah virus (NiV) is a deadly zoonotic pathogen with a high mortality rate and no approved vaccines, creating an urgent need for new solutions. We developed a novel, thermostable, orally administered NiV vaccine using a human serotype 5 adenovirus (AdHu5) vector to express NiV glycoproteins G and F. This vaccine, delivered via an enteric-coated capsule using the OraPro platform, eliminates the cold chain and allows for self-administration. In a Syrian hamster model, a prime-and-pull strategy—an intramuscular prime followed by an oral booster—provided 100% protection against a lethal NiV challenge, significantly outperforming intramuscular administration alone, which offered only 66% protection. The oral booster also led to higher serum neutralizing antibody titres while reducing anti-vector immunity, potentially enabling repeated use of the same vector. Histopathological analysis showed superior protection in orally boosted animals, with minimal lung lesions. This study highlights the potential of oral vaccines for addressing emerging infectious diseases.
Data availability
Data presented in this study are described in the Supplementary Materials, or available upon request. Nucleotide sequences are available at GenBank accession numbers PX246253, PX246254 and PX246255 at https://www.ncbi.nlm.nih.gov/genbank/.
Code availability
R code used for logistic regression is available in the Supplementary Materials.
References
Soni, M., Kumar, V., Singh, M. P., Shabil, M. & Sah, S. Nipah virus resurgence: a call for preparedness across states. Infect. Med. 3 (4), 100145 (2024).
Olatunji, G., Kokori, E., Abdulrahmon, M. A. & Aderinto, N. Addressing the recurrent Nipah Virus outbreaks: A call for vigilance, collaboration, and preparedness. New Microb. New Infect. 54, 101184 (2023).
Gazal, S. et al. Nipah and hendra viruses: Deadly zoonotic paramyxoviruses with the potential to cause the next pandemic. Pathog. Basel Switz. 11 (12), 1419 (2022).
Singh, R. K. et al. Nipah virus: epidemiology, pathology, immunobiology and advances in diagnosis, vaccine designing and control strategies - a comprehensive review. Vet. Q. 39 (1), 26–55 (2019).
Playford, E. G. et al. Safety, tolerability, pharmacokinetics, and immunogenicity of a human monoclonal antibody targeting the G glycoprotein of henipaviruses in healthy adults: a first-in-human, randomised, controlled, phase 1 study. Lancet Infect. Dis. 20 (4), 445–454 (2020).
Moon, S. Y. et al. Immunogenicity and neutralization of recombinant vaccine candidates expressing F and G glycoproteins against Nipah virus. Vaccines 12 (9), 999 (2024).
Huang, X. et al. Nipah virus attachment glycoprotein ectodomain delivered by type 5 adenovirus vector elicits broad immune response against NiV and HeV. Front. Cell Infect. Microbiol. https://doi.org/10.3389/fcimb.2023.1180344/full (2023).
Lu, M. et al. Both chimpanzee adenovirus-vectored and DNA vaccines induced long-term immunity against Nipah virus infection. NPJ Vaccines 8 (1), 1–12 (2023).
van Doremalen, N. et al. A single-dose ChAdOx1-vectored vaccine provides complete protection against Nipah Bangladesh and Malaysia in Syrian golden hamsters. PLoS Negl. Trop. Dis. 13 (6), e0007462 (2019).
Lu, M. et al. Single-dose intranasal AdC68-vectored vaccines rapidly protect Syrian hamsters against lethal Nipah virus infection. Mol. Ther. J. Am. Soc. Gene Ther. 33 (7), 3270–3285 (2025).
Woolsey, C. et al. Recombinant vesicular stomatitis virus-vectored vaccine induces long-lasting immunity against Nipah virus disease. J. Clin. Invest. 133 (3), e164946 (2023).
Chen, S. et al. Ferritin nanoparticle-based Nipah virus glycoprotein vaccines elicit potent protective immune responses in mice and hamsters. Virol. Sin. S1995-802X (24), 00144–00145 (2024).
Zhou, D. et al. An attachment glycoprotein nanoparticle elicits broadly neutralizing antibodies and protects against lethal Nipah virus infection. NPJ Vaccines 9 (1), 158 (2024).
Welch, S. R. et al. Single-dose mucosal replicon-particle vaccine protects against lethal Nipah virus infection up to 3 days after vaccination. Sci. Adv. 9 (31), eadh4057 (2023).
Pedrera, M. et al. Evaluation of the immunogenicity of an mRNA vectored Nipah virus vaccine candidate in pigs. Front. Immunol. 15, 1384417 (2024).
Findlay-Wilson, S. et al. Establishment of a Nipah virus disease model in hamsters, including a comparison of intranasal and intraperitoneal routes of challenge. Pathog. Basel Switz. 12 (8), 976 (2023).
Vela Ramirez, J. E., Sharpe, L. A. & Peppas, N. A. Current state and challenges in developing oral vaccines. Adv. Drug Deliv. Rev. 114, 116–131 (2017).
Bernstein, D. I. et al. Successful application of prime and pull strategy for a therapeutic HSV vaccine. NPJ Vaccines. 4 (1), 1–10 (2019).
Shin, H. & Iwasaki, A. A vaccine strategy that protects against genital herpes by establishing local memory T cells. Nature 491 (7424), 463–467 (2012).
Drew J. Virus. US12016949B2, https://patents.google.com/patent/US12016949B2/en?oq=Drew+OraPro+US12016949 (2024).
Bacon, A. et al. Generation of a thermostable, oral Zika vaccine that protects against virus challenge in non-human primates. Vaccine S0264-410X (23), 00198 (2023).
Bochkov, Y. A. & Palmenberg, A. C. Translational efficiency of EMCV IRES in bicistronic vectors is dependent upon IRES sequence and gene location. BioTechniq. 41 (3), 283–284 (2006).
Kim, J. H. et al. High cleavage efficiency of a 2A peptide derived from porcine teschovirus-1 in human cell lines, zebrafish and mice. PLoS ONE 6 (4), e18556 (2011).
Bossart, K. N. et al. Receptor binding, fusion inhibition, and induction of cross-reactive neutralizing antibodies by a soluble G glycoprotein of Hendra virus. J. Virol. 79 (11), 6690–6702 (2005).
Martin, L. H., Calabi, F., Lefebvre, F. A., Bilsland, C. A. & Milstein, C. Structure and expression of the human thymocyte antigens CD1a, CD1b, and CD1c. Proc. Natl. Acad. Sci. 84 (24), 9189–9193 (1987).
Witting, S. R., Vallanda, P. & Gamble, A. L. Characterization of a third generation lentiviral vector pseudotyped with Nipah virus envelope proteins for endothelial cell transduction. Gene Ther. 20 (10), 997–1005 (2013).
Luo, X. et al. Establishment of a neutralization assay for Nipah virus using a high-titer pseudovirus system. Biotechnol. Lett. 45 (4), 489–498 (2023).
Berkhout, B. A fourth generation lentiviral vector: Simplifying genomic gymnastics. Mol. Ther. 25 (8), 1741–1743 (2017).
Pernet, O., Pohl, C., Ainouze, M., Kweder, H. & Buckland, R. Nipah virus entry can occur by macropinocytosis. Virology 395 (2), 298–311 (2009).
Scallan, C. D., Tingley, D. W., Lindbloom, J. D., Toomey, J. S. & Tucker, S. N. An adenovirus-based vaccine with a double-stranded RNA adjuvant protects mice and ferrets against H5N1 avian influenza in oral delivery models. Clin. Vaccine Immunol. CVI. 20 (1), 85–94 (2013).
Chan, H. Y. et al. Comparison of IRES and F2A-based locus-specific multicistronic expression in stable mouse lines. PLoS ONE 6 (12), e28885 (2011).
Kuzmich, A. I., Vvedenskii, A. V., Kopantzev, E. P. & Vinogradova, T. V. Quantitative comparison of gene co-expression in a bicistronic vector harboring IRES or coding sequence of porcine teschovirus 2A peptide. Russ. J. Bioorg. Chem. 39 (4), 406–416 (2013).
Streeter, P. R., Berg, E. L., Rouse, B. T., Bargatze, R. F. & Butcher, E. C. A tissue-specific endothelial cell molecule involved in lymphocyte homing. Nature 331 (6151), 41–46 (1988).
Shoji, M., Yoshizaki, S., Mizuguchi, H., Okuda, K. & Shimada, M. Immunogenic comparison of chimeric adenovirus 5/35 vector carrying optimized human immunodeficiency virus clade C genes and various promoters. PLoS ONE 7 (1), e30302 (2012).
Forrest, B. D., LaBrooy, J. T., Robinson, P., Dearlove, C. E. & Shearman, D. J. Specific immune response in the human respiratory tract following oral immunization with live typhoid vaccine. Infect. Immun. 59 (3), 1206–1209 (1991).
Brandtzaeg, P. Do salivary antibodies reliably reflect both mucosal and systemic immunity?. Ann. N. Y. Acad. Sci. 1098, 288–311 (2007).
Farstad, I. N. et al. Human intestinal B-cell blasts and plasma cells express the mucosal homing receptor integrin alpha 4 beta 7. Scand. J. Immunol. 42 (6), 662–672 (1995).
Zak, D. E. et al. Merck Ad5/HIV induces broad innate immune activation that predicts CD8+ T-cell responses but is attenuated by preexisting Ad5 immunity. Proc. Natl. Acad. Sci. USA 109 (50), E3503-3512 (2012).
Chang, Y. Y., Enninga, J. & Stévenin, V. New methods to decrypt emerging macropinosome functions during the host-pathogen crosstalk. Cell Microbiol. 23 (7), e13342 (2021).
Krzyzaniak, M. A., Zumstein, M. T., Gerez, J. A., Picotti, P. & Helenius, A. Host cell entry of respiratory syncytial virus involves macropinocytosis followed by proteolytic activation of the F protein. PLoS Pathog. 9 (4), e1003309 (2013).
Plotkin, S. A. Recent updates on correlates of vaccine-induced protection. Front. Immunol. https://doi.org/10.3389/fimmu.2022.1081107 (2023).
Escudero-Pérez, B., Lawrence, P. & Castillo-Olivares, J. Immune correlates of protection for SARS-CoV-2, Ebola and Nipah virus infection. Front. Immunol. 14, 1156758 (2023).
Travieso, T., Li, J., Mahesh, S., Mello, J. D. F. R. E. & Blasi, M. The use of viral vectors in vaccine development. NPJ Vaccines 7 (1), 75 (2022).
Abrignani, M. G. et al. COVID-19, vaccines, and thrombotic events: A narrative review. J. Clin. Med. 11 (4), 948 (2022).
Nicolai, L. et al. Thrombocytopenia and splenic platelet-directed immune responses after IV ChAdOx1 nCov-19 administration. Blood 140 (5), 478–490 (2022).
Sejvar, J. J. et al. Long-term neurological and functional outcome in Nipah virus infection. Ann. Neurol. 62 (3), 235–242 (2007).
Liew, Y. J. M. et al. The immunobiology of Nipah virus. Microorganisms 10 (6), 1162 (2022).
Afkhami, S., Yao, Y. & Xing, Z. Methods and clinical development of adenovirus-vectored vaccines against mucosal pathogens. Mol. Ther. Methods Clin. Dev. 3, 16030 (2016).
Crawford, K. H. D. et al. Protocol and reagents for pseudotyping lentiviral particles with SARS-CoV-2 spike protein for neutralization assays. Viruses 12 (5), 513 (2020).
Ferrara, F. & Temperton, N. Pseudotype neutralization assays: From laboratory bench to data analysis. Methods Protoc. 1 (1), 8 (2018).
Acknowledgements
The study was supported by Innovate UK contract 10027019 and iosBio Ltd.
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JD, AB, EJB designed the programme of work. JD, PB designed the vectors, MS, AB, GE, LC, LE, SD, SFW, SF, RH, EK, IRT, JS conducted the experiments. MS, CL, JS, IRT, AB, SD analysed data and prepared figures. CL, EJB, SD and JD wrote the manuscript. All authors reviewed the final manuscript.
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PB, CL, GE, LC, AB and EJB are employees of iosBio Ltd. MS and JD are former employees of iosBio Ltd. PB, AB, EJB, JD and CL hold share options or shares of iosBio Ltd. LE, SD, SFW, SF, EK, IRT, JS declare no conflict.
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Stewart, M., Bone, P., Bacon, A. et al. Protective immunity in hamsters from an oral Nipah vaccine correlates with pseudovirus neutralising antibody titre. Sci Rep (2026). https://doi.org/10.1038/s41598-026-40205-2
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DOI: https://doi.org/10.1038/s41598-026-40205-2
