采用流式细胞仪检测艾滋病病毒1型膜蛋白小鼠免疫后IFN-γ的表达

Detection of IKN-r Expression of Mice by Flow Cytometry Primed with DNA and Boosted with Kecombinant Vaccinia virus Expressing Gpl20 gene of Human Immunodeficiency Virus

目的通过流式细胞技术检测艾滋病病毒1型(HIV-1)Gp120膜蛋白免疫小鼠后的IFN-γ的表达。方法 构建表达HIV-1gp120基因的复制型重组痘苗病毒。与DNA疫苗联合免疫小鼠,用流式细胞仪检测小鼠脾淋巴细胞中IFN-γ的表达。结果 获得表达中国HIV-1流行株gp120基因的重组痘苗病毒rVVgp120、rVVM304,能正确表达Gp120蛋白。与DNA疫苗p120-VRC、pM304-VRC联合免疫小鼠后,用流式细胞仪检测到小鼠脾淋巴细胞能特异性的表达IFN-γ。结论 用流式细胞检测技术成功检测到免疫小鼠的脾淋巴细胞特异性的表达IFN-γ,用此方法可方便快捷地检测小鼠的细胞免疫效果。

Objective To detect IFN-y expression of mice immunized with DNA and recombinant vaccinia expressing gp120gene of human immunodeficiency virus by using flow cytometry. Method Replication-competent recombinant vaccinia virus rVVgp120 and rVVM304 expressing gp120 and M304 gene was constructed respectively. Its immunogenicity was evaluated by testing IFN-y levels of mice primed with DNA vaccines p120-VRC, pM304-VRC and boosted with recombinant vaccinia through flow cytometry. Results The replication-competent recombinant vaccinia can virus express gp120 protein correctly. The mice primed with DNA vaccines and boosted with recombinant vaccinia virus elicited HIV-1 specific IFN-y.Conclusion Flow cytometry can detect mouse IFN-y immunized with DNA and recombinant vaccinia virus expressing gp120 gene of human immunodeficiency virus. It is a rapid method to detect the cellular immune responses.