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Optimization of storage conditions for Puumala virus based vaccine

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1. Title Title of document Optimization of storage conditions for Puumala virus based vaccine
2. Creator Author's name, affiliation, country Anna N. Vetrova; Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of the Russian Academy of Sciences; Sechenov First Moscow State Medical University (Sechenov University); Россия
2. Creator Author's name, affiliation, country Svetlana S. Kurashova; Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of the Russian Academy of Sciences; Россия
2. Creator Author's name, affiliation, country Rostislav D. Teodorovich; Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of the Russian Academy of Sciences; Россия
2. Creator Author's name, affiliation, country Yulia V. Popova; Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of the Russian Academy of Sciences; Россия
2. Creator Author's name, affiliation, country Ekaterina A. Blinova; Central Research Institute of Epidemiology of The Federal Service on Customers’ Rights Protection and Human Well-being Surveillance; Россия
2. Creator Author's name, affiliation, country Pavel A. Nabatnikov; Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of the Russian Academy of Sciences; Россия
2. Creator Author's name, affiliation, country Evgeny А. Tkachenko; Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of the Russian Academy of Sciences; Россия
2. Creator Author's name, affiliation, country Tamara K. Dzagurova; Chumakov Federal Scientific Center for Research and Development of Immune-and-Biological Products of the Russian Academy of Sciences; Россия
3. Subject Discipline(s)
3. Subject Keyword(s) hemorrhagic fever with renal syndrome; Puumala virus; hantavirus vaccine; neutralizing antibodies; immune response; human serum albumin
4. Description Abstract

We report the effects of storage conditions and human serum albumin addition to the vaccine composition on the immunogenic properties of an experimental vaccine against hemorrhagic fever with renal syndrome. To obtain an experimental vaccine, the Puumala virus, propagated in Vero cells culture, was concentrated, inactivated with beta-propiolactone, and purified by chromatography on the Capto™Core 700 multimodal sorbent (GE Healthcare). The target component in the vaccine was 2±0.2 × 106 of viral RNA copies/ml. The humoral immune response to the vaccine was determined by measuring the neutralizing antibodies in the blood serum of immunized Syrian hamsters (Mesocricetus auratus). It was revealed that the Puumala virus inactivated with beta-propiolactone induces a pronounced humoral immune response, which indicates preservation of relevant immunogenic epitopes. Аccording to our study, viral RNA is more stable during storage of the inactivated vs. intact virus. It was confirmed that full virus inactivation occurred after 24 hours of storage at 37°C. A significant decrease in the virus immunogenicity during the thermal inactivation was noted, which confirms the need for careful selection of storage conditions for thermolabile Hantaviruses. A direct correlation was observed between viral titer and viral RNA copy number/ml. Adding human serum albumin stabilized viral infectivity during storage: Puumala virus titer remained at the same level for 3 months of storage at 6±2°C, whereas without albumin, the titer decreased to undetectable level. The addition of 0.1% human serum albumin to the inactivated virus stabilized its immunogenic properties during long-term storage. It was shown that vaccine preparations with human serum albumin (0.1–1%) show higher dose-dependent stability upon repeated freezing assessed by detecting viral RNA level and their immunogenicity. It can be assumed that human serum albumin provides better preservation of of viral genetic material during storage, as well the protein epitopes responsible for the protective immune response in the form of neutralizing antibodies induction, undergo less pronounced conformational changes in the presence of albumin. These data support the addition of human serum albumin to the vaccine formulation.

5. Publisher Organizing agency, location SPb RAACI
6. Contributor Sponsor(s)
7. Date (DD-MM-YYYY) 24.04.2023
8. Type Status & genre Peer-reviewed Article
8. Type Type Short Communication
9. Format File format PDF (Rus),
10. Identifier Uniform Resource Identifier https://iimmun.ru/iimm/article/view/2116
10. Identifier Digital Object Identifier (DOI) 10.15789/2220-7619-OOS-2116
11. Source Title; vol., no. (year) Russian Journal of Infection and Immunity; Vol 13, No 2 (2023)
12. Language English=en ru
13. Relation Supp. Files Figure 1. A relation between the stability of non-inactivated Puumala virus (PUU) and storage conditions (314KB)
Figure 2. A stability analysis for Puumala virus inactivated by beta-propiolactone (I-PUU) (163KB)
Figure 3. A relation between the immunogenicityof beta-propiolactone inactivated hantavirus vaccine and storage conditionsNote. The neutralizing antibody titer (n = 6) were determined in Syrian hamsters blood sera after dual immunization in undiluted form: storage for 3 months at 6±2°C without additives (I- PUU); storage for 3 months at 6±2°C with 0.1% HSA (I-PUU × 0.1) or after freezing 5 times (I-PUU-5f × 0.1), or after freezing 40 times (I-PUU-40f × 0.1); storage for 6 days at 37°C (I-PUU-37). The neutralizing antibodies titer (NAb) was determined by the focus reduction neutralization test (FRNT50) using Vero cells. (76KB)
14. Coverage Geo-spatial location, chronological period, research sample (gender, age, etc.)
15. Rights Copyright and permissions Copyright (c) 2023 Vetrova A.N., Kurashova S.S., Teodorovich R.D., Popova Y.V., Blinova E.A., Nabatnikov P.A., Tkachenko E.А., Dzagurova T.K.
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