RESUMEN
To study the immunogenicity of recombinant adeno-asscociated virus type 1 expressing HIV-1 gp120 gene (rAAV2/1-gp120) in BALB/c mice and Rhesus macaques. The gp120 gene derived from Chinese HIV-1 isolates was constructed into rAAV2/1 and rAd5 vectors. Firstly, the immunogenicity of rAAV2/1-gp120 was compared with rAd5-gp120 in BALB/c mice when used once or twice in 3 weeks interval. Then the monkeys were immunized with rAAV2/1-gp120 once. The HIV-1 specific IgG levels and neutralization activity to pseudotyped HIV-1 virus were tested using ELISA and neutralization assay, and the cellular immune responses were analyzed by IFN-gamma enzyme-linked immunospot (ELISPOT) and in vivo CTL assays. Compared with rAd5-gp120 immunized mice, mice immunized with rAAV2/1-gp120 once in duced stronger gp120-specific IgG and were sustained for at least 21 weeks. rAd5-gp120 immunized mice generated stronger cellular immune responses than rAAV2/1-gp120 in spleen and draining lymph node. But only moderate gp120-specific in vivo CTL activity was observed in both rAAV2/1-gp120 and rAd5-gp120 immunized mice. Four of five monkeys vaccinated with rAAV2/1-gp120 generated gp120 specific IgG, the titer ranged from 1:100 to 1:400 with end-point dilution. Gp120 specific IgG could be detected 4 weeks after immunization and reached the peak at 10 weeks after immunization. No neutralization activity against pseudotyped HIV-1 virus expressing NL4-3 Env antigen was detected. In Conclusion, rAAV2/1-gp120 induced high level of HIV-1 specific IgG antibody and moderate cellular immune responses. No neutralizing antibody was elicited. It indicates that the env gene and immunization strategy should be optimized to elicit neutralizing antibody against HIV-1 in further studies.
Asunto(s)
Anticuerpos Anti-VIH/inmunología , Proteína gp120 de Envoltorio del VIH/inmunología , VIH-1/inmunología , Proteínas Recombinantes de Fusión/inmunología , Vacunas contra el SIDA/administración & dosificación , Vacunas contra el SIDA/inmunología , Adenoviridae/genética , Animales , Células COS , Chlorocebus aethiops , Citotoxicidad Inmunológica/inmunología , Dependovirus/genética , Ensayo de Inmunoadsorción Enzimática , Vectores Genéticos/genética , Proteína gp120 de Envoltorio del VIH/genética , VIH-1/genética , Inmunización/métodos , Inmunoglobulina G/inmunología , Macaca mulatta , Ratones , Ratones Endogámicos BALB C , Proteínas Recombinantes de Fusión/administración & dosificación , Proteínas Recombinantes de Fusión/genética , Factores de TiempoRESUMEN
OBJECTIVE: To construct DNA and recombinant adenovirus vector vaccines containing an env gene from the prevalent subtype B strain in China and try to use them for therapeutic and prophylactic vaccines. METHODS: The candidate plasmid DNA vaccine pVR-gp160 and recombinant adenovirus vaccine rAdV-gp160 were constructed separately. BALB/c mice were immunized with these two vaccines in different administration schemes. HIV-1 Gp120-specific cellular responses and antibody levels were detected by ELISPOT and ELISA respectively. RESULTS: DNA vaccine alone and combined vaccines in a DNA prime/rAdV-gp160 boost vaccination regimen induced high level of Gp120-specific cellular responses. While low level of Gp120-specific antibodies were elicited in all groups. CONCLUSION: DNA and rAdV vaccines could efficiently express Gp160 protein and activate specific cellular responses.
Asunto(s)
Vacunas contra el SIDA/inmunología , Adenoviridae/inmunología , Genes env/inmunología , Vectores Genéticos/inmunología , VIH-1/inmunología , Plásmidos/inmunología , Vacunas de ADN/inmunología , Vacunas contra el SIDA/genética , Adenoviridae/genética , Animales , China , Vectores Genéticos/genética , Anticuerpos Anti-VIH/genética , Anticuerpos Anti-VIH/inmunología , Proteína gp120 de Envoltorio del VIH/genética , Proteína gp120 de Envoltorio del VIH/inmunología , Proteínas gp160 de Envoltorio del VIH/genética , Proteínas gp160 de Envoltorio del VIH/inmunología , VIH-1/genética , Ratones , Ratones Endogámicos BALB C , Plásmidos/genética , Vacunas de ADN/genética , Vacunas Sintéticas/genética , Vacunas Sintéticas/inmunologíaRESUMEN
OBJECTIVE: To compare the immunogenicity of rAAV2/1 and rAd5 expressing HIV-1 gag in BALB/c mice. METHODS: BALB/c mice were immunized with rAAV2/1-gag or rAd5-gag once or twice. HIV-1 specific cellular immune responses were analyzed by in vivo CTL and intracellular cytokine staining assays. HIV-1 Gag specific antibodies were tested by ELISA. RESULTS: Mice immunized with rAd5-gag once induced stronger Gag specific cellular immune responses and similar level of Gag specific antibody compared with rAAV2/1-gag. Mice immunized with rAd5-gag reached the peak immune responses more rapidly than rAAV2/1-gag. However, mice immunized with rAAV2/1-gag twice elicited better Gag specific IgG. CONCLUSION: rAd5-gag induced strong HIV-1 specific cellular and antibody responses, and rAAV2/1-gag induced high level of HIV-1 specific IgG and moderate cellular immune responses.