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Study of the protective properties of immunodominant proteins of orthopoxviruses in various methods of immunization

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1. Title Title of document Study of the protective properties of immunodominant proteins of orthopoxviruses in various methods of immunization
2. Creator Author's name, affiliation, country Denis I. Zrelkin; National Research Center of Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya; Россия
2. Creator Author's name, affiliation, country Olga V. Zubkova; National Research Center of Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya; Россия
2. Creator Author's name, affiliation, country Tatiana A. Ozharovskaia; National Research Center of Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya; Россия
2. Creator Author's name, affiliation, country Olga Popova; National Research Center of Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya; Россия
2. Creator Author's name, affiliation, country Daria V. Voronina; National Research Center of Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya; Россия
2. Creator Author's name, affiliation, country Polina P. Goldovskaya; National Research Center of Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya; Россия
2. Creator Author's name, affiliation, country Irina V. Vavilova; National Research Center of Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya; Россия
2. Creator Author's name, affiliation, country Anastasia S. Chugunova; National Research Center of Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya; Россия
2. Creator Author's name, affiliation, country Inna V. Dolzhikova; National Research Center of Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya; Россия
2. Creator Author's name, affiliation, country Dmitry N. Shcherbinin; National Research Center of Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya; Россия
2. Creator Author's name, affiliation, country Dmitry V. Shcheblyakov; National Research Center of Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya; Россия
2. Creator Author's name, affiliation, country Denis Y. Logunov; National Research Center of Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya; Россия
2. Creator Author's name, affiliation, country Alexander L. Gintsburg; National Research Center of Epidemiology and Microbiology named after Honorary Academician N.F. Gamaleya; Россия
3. Subject Discipline(s)
3. Subject Keyword(s) monkeypox virus; vaccinia virus; recombinant adenovirus; immunogenicity; protective activity
4. Description Abstract

Introduction. In 2022, the World Health Organization declared monkeypox a public health emergency. The monkeypox virus (MPV) is part of the Orthopoxvirus genus within the Poxviridae family. During replication, orthopoxviruses produce two distinct forms of viral particles: the extracellular enveloped virion (EEV), released via exocytosis, and the intracellular mature virion (IMV), released through cell lysis. These forms differ in surface proteins composition, influencing their immunogenicity and infectivity.

Aim. To evaluate the immunogenic and protective activity of nine surface antigens of vaccinia virus.

Materials and methods . Recombinant human adenoviruses type 2 (rAd2) carrying surface antigens of vaccinia virus were obtained using homologous recombination in bacteria, followed by adenoviral particle assembly in HEK293 cells. The immunogenic and protective properties of these adenoviruses were tested in BALB/c mice. The presence of antibodies to the vaccinia virus was assessed using ELISA, and survival rates were evaluated in a lethal infection model after intranasal challenge with the vaccinia virus strain Western Reserve.

Results. The most immunogenic and protective antigens of the vaccinia virus within rAd2 were glycoprotein B5 of the EEV and membrane-associated protein H3 of the IMV, both showing 100% protective efficacy after intranasal immunization.

Conclusion. Using a panel of recombinant adenoviruses carrying genes of vaccinia virus surface proteins, it was shown that optimal protection is achieved using a combination of enveloped and mature virion antigens. This method could be used for development of new multivalent preparations against various viral infections.
5. Publisher Organizing agency, location Central Research Institute for Epidemiology
6. Contributor Sponsor(s)
7. Date (DD-MM-YYYY) 30.03.2025
8. Type Status & genre Peer-reviewed Article
8. Type Type Research Article
9. Format File format
10. Identifier Uniform Resource Identifier https://microbiol.crie.ru/jour/article/view/18755
10. Identifier Digital Object Identifier (DOI) 10.36233/0372-9311-643
10. Identifier eLIBRARY Document Number (EDN) ONKJVE
11. Source Title; vol., no. (year) Journal of microbiology, epidemiology and immunobiology; Vol 102, No 1 (2025)
12. Language English=en en
13. Relation Supp. Files Fig. 1. Scheme for the production of recombinant adenoviruses expressing surface antigens of the vaccinia virus. L1 — myristoylated protein; H3 — membrane-associated protein p35 IMV; D8 — transmembrane carbonic anhydrase-like protein; D13 — IMV membrane protein; A14 — structural transmembrane protein p16 IMV; A17 — transmembrane protein IMV (morphogenesis factor); A27 — membrane protein IMV; B5 — type I membrane glycoprotein; A33 — type II membrane glycoprotein. (208KB)
Fig. 2. Immunogenicity of recombinant adenoviruses expressing surface antigens of the vaccinia virus after single (a) and double (b) intramuscular immunization. The figures show the confidence at p < 0.05; *p < 0.0001. C — control. (136KB)
Fig. 3. Immunogenicity of recombinant adenoviruses expressing surface antigens of the vaccinia virus after two intranasal immunizations. The figure shows the confidence at p < 0.05; *p < 0.0001. C– — control. (60KB)
Fig. 4. The protective efficacy of recombinant adenovirus vectors expressing the surface antigens of the vaccinia virus after intramuscular (a, b) or intranasal (c, d) immunization in a mouse model of lethal infection. a, c — weight dynamics (the graph shows the average value and the standard error of the average value); b, d — survival. C– — control. (187KB)
Fig. 5. The protective efficacy of recombinant adenovirus vectors rAd2-B5 and rAd2-H3 after combined immunization (im+in) in a mouse model of lethal infection. a — weight dynamics; b — survival. C — control. (74KB)
14. Coverage Geo-spatial location, chronological period, research sample (gender, age, etc.)
15. Rights Copyright and permissions Copyright (c) 2025 Zrelkin D.I., Zubkova O.V., Ozharovskaia T.A., Popova O., Voronina D.V., Goldovskaya P.P., Vavilova I.V., Chugunova A.S., Dolzhikova I.V., Shcherbinin D.N., Shcheblyakov D.V., Logunov D.Y., Gintsburg A.L.
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