Abstract
Smallpox eradication and lack of adequate animal model for smallpox infection underlies a necessity to assess immunogenic and protective properties of genetic engineering-created live attenuated smallpox vaccines in several animal models of orthopoxviral infections. Here we compared immunogenic and protective properties of the recombinant vaccinia virus (VACV) LIVP-GFP intradermally (i.d.) inoculated to mice, guinea pigs and rabbits. LIVP-GFP immunization in all animal species was applied at dose of 2 × 104 or 2 × 106 PFU. Control animals were injected with saline. Blood sampling was performed on day 28 after virus LIVP-GFP or saline inoculation. Blood samples were taken intravitally from the retro-orbital venous sinus in mice, heart in guinea pigs or marginal ear vein in rabbits. Serum samples were isolated by precipitating blood cells via centrifugation. The serum anti-VACV IgG titers were determined by ELISA. On day 30 post-immunization animals were intranasally challenged with lethal dose of host specific orthopoxvirus species. Mice were infected by cowpox virus (CPXV) strain GRI-90 at dose 68 LD50, guinea pigs – by VACV GPA at dose 56 LD50, rabbits — by VACV HB-92 at dose 100 LD50. All animals in control group died afterwards, whereas all animals immunized by attenuated recombinant virus LIVP-GFP at dose 2 × 106 PFU survived. In case of the LIVP-GFP immunization at dose 2 × 104 PFU, 88% of mice, 67% of rabbits and 50% of guinea pigs survived after being challenged with species-specific CPXV, VACV HB-92, and VACV GPA. ELISA data for the blood serum samples revealed a correlation between level of VACV-specific antibodies and level of protection in animal species. Based on the data obtained, it could be concluded that all three “animal–orthopoxvirus” models allow to provide with a proper evaluation of immunogenicity and protectiveness for generated modern attenuated vaccines against smallpox and other orthopoxviral human infections. Upon that, it was shown that BALB/c mouse strain was the most convenient investigational host species.
Highlights
Вирус осповакцины (Vaccinia virus, vaccinia virus (VACV)) стал живой противооспенной вакциной, с помощью которой впервые удалось ликвидировать особо опасное инфекционное заболевание человека [5]
Data for animal groups immunized with virus LIVPGFP or groups unimmunized with LIVP-GFP and uninfected or infected by corresponding lethal species-specific orthopoxvirus
Определенные методом ИФА титры vaccinia virus (VACV)-специфичных антител в сыворотке крови мышей, морских свинок и кроликов, инфицированных VACV LIVP-GFP в дозах 104 БОЕ (А) и 106 БОЕ (Б) Figure 2
Summary
ФБУН Государственный научный центр вирусологии и биотехнологии «Вектор» Роспотребнадзора, р.п. В данной работе проведено сравнение иммуногенных и протективных свойств рекомбинантного вируса осповакцины (VACV) LIVP-GFP при внутрикожном (в/к) введении мышам, морским свинкам и кроликам. Иммунизирующие дозы вируса LIVP-GFP для всех видов животных составляли 2 × 104 или 2 × 106 БОЕ. Забор проб крови у животных проводили прижизненно через 28 суток после введения вируса LIVP-GFP или физиологического раствора: у мышей — из ретроорбитального венозного синуса, у морских свинок — из сердца, у кроликов — из краевой вены уха. Мышей заражали вирусом оспы коров (CPXV) GRI-90 в дозе 68 ЛД50, морских свинок — VACV GPA в дозе 56 ЛД50, кроликов — VACV НВ-92 в дозе 100 ЛД50. При иммунизирующей дозе LIVP-GFP 2 × 104 БОЕ выжило 88% мышей после заражения CPXV, 67% кроликов после инфекции VACV НВ-92 и 50% морских свинок после заражения VACV GPA.
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