Effect of the ati Gene Deletion on the Pathogenicity and Immunogenicity of the Vaccinia Virus

S N Yakubitskiy; A A Sergeev; K A Titova; I S Shulgina; E V Starostina; M B Borgoyakova; L I Karpenko; S N Shchelkunov

doi:10.32607/actanaturae.17872

Effect of the ati Gene Deletion on the Pathogenicity and Immunogenicity of the Vaccinia Virus

Acta Naturae. 2023 Jul-Sep;15(3):82-90. doi: 10.32607/actanaturae.17872.

Authors

S N Yakubitskiy¹, A A Sergeev¹, K A Titova¹, I S Shulgina¹, E V Starostina¹, M B Borgoyakova¹, L I Karpenko¹, S N Shchelkunov¹

Affiliation

¹ State Research Center of Virology and Biotechnology VECTOR, Rospotrebnadzor, Koltsovo, Novosibirsk region, 630559 Russian Federation.

Abstract

Among the nonvirion proteins of the vaccinia virus (VACV), a 94-kDa long protein is most abundantly present; the protein is a truncated form of the 150-kDa A-type inclusion (ATI) protein of the cowpox virus encoded by the ati gene. This VACV protein does not form intracellular ATIs, being as it is a major immunogen upon infection/immunization of humans or animals with the VACV. Antibodies specific to this protein are not virus-neutralizing. The present study focused on the effect of the production of this nonstructural major immunogenic VACV protein on the manifestation of pathogenicity and immunogenicity of the virus in the BALB/c mouse model of infection. In order to introduce a targeted deletion into the VACV LIVP genome, the recombinant integration/deletion plasmid pΔati was constructed and further used to generate the recombinant virus LIVPΔati. The pathogenicity of the VACV LIVP and LIVPΔati strains was studied in 3-week-old mice. The mice were intranasally infected with the viruses at a dose of 107 pfu; 50% of the animals infected with the parent LIVP strain died, while infection with the LIVPΔati strain led to the death of only 20% of the mice. Intradermal vaccination of mice aged 6- weeks with the LIVPΔati virus statistically significantly increased the production of VACV-specific IgG, compared to that after intradermal vaccination with VACV LIVP. Meanwhile, no differences were noted in the cell-mediated immune response to the vaccination of mice with VACV LIVP or LIVPΔati, which was assessed by ELISpot according to the number of splenocytes producing IFN-γ in response to stimulation with virus-specific peptides. Intranasal infection of mice with lethal doses of the cowpox virus or the ectromelia virus on day 60 post-immunization with the studied VACV variants demonstrated that the mutant LIVPΔati elicits a stronger protective response compared to the parent LIVP.

Keywords: ati gene; immunogenicity; intradermal injection; orthopoxviruses; protectivity; vaccinia virus.