负载Ad5-HIVgag/env的DC疫苗在小鼠体内的免疫原性研究

Immunogenicity of dendritic cells pulsed with Ad5-HIVgag/env in mice

目的 用Ad5-HIV gag/env负载小鼠树突状细胞（Dendritic cell,DC）,探索其在小鼠体内诱导的HIV Gag/Env特异性细胞免疫应答.方法 贴壁法分离纯化BALB/c小鼠DCs,并用细胞因子诱导DC细胞成熟,采用流式细胞术鉴定其纯度.利用重组腺病毒Ad5-HIV gag/env感染DC细胞,制备DC疫苗.用所制备DC疫苗免疫小鼠,在免疫后的不同时间点,用ELISPOT方法检测小鼠体内HIV特异性细胞免疫应答水平.结果 成功培养出Ad5-HIVgag/env负载的DC疫苗;免疫荧光以及流式细胞术检测到Gag/Env抗原可在DC内有效表达;上述DC疫苗免疫小鼠后能诱导高水平HIV特异性细胞免疫反应.结论 Ad5-HIV gag/env负载的树突状细胞疫苗可以在小鼠体内诱导较强的HIV特异性细胞免疫反应.

Objective To evaluate HIV Gag/Env specific cellular immune responses induced in mice immunized with mouse dendritic cells pulsed with Ad5-HIVgag/env.Methods BALB/c mouse DCs were generated by culture of adherent bone marrow cells in the presence of several cytokines.The purity of matured DCs was determined by flow cytometry.DC vaccines were prepared by loading them with replication-deficient recombinant adenovirus Ad5-HIVgag or Ad5-HIVenv.BALB/c mice were immunized by intramuscular injection of above DC vaccines alone or in combination,at different time points post immunization HIV specific cellular immune responses detected by ELISPOT assay.Results Mouse DC vaccines loaded with Ad5-HIVgag/env were generated successfully.Expression of HIV Gag or Env antigens in DC vaccines could be detected by immunofluorescence assay and flow cytometry;and high levels of HIV-specific cellular immune responses were induced in mice immunized with above DC vaccines Conclusions Dendritic cells loaded with Ad5-HIVgag/env could elicit potent HIV specific cellular immune responses in mice.