HIV疫苗小鼠免疫原性检测方法的初步建立

Development of A Method for Determination of Immunogenicity of HIV Vaccine in Mice

目的初步建立HIV疫苗小鼠免疫原性检测方法,并对HIV疫苗进行免疫原性评价。方法选择一种HIVDNA疫苗(D)、2种重组痘苗载体HIV疫苗(M和R),分别免疫BALB/c小鼠,6周后分离血清和脾淋巴细胞,采用ELISOPT和胞内因子染色法检测细胞因子分泌情况,MTS法检测淋巴细胞增殖反应,ELISA法检测血清中抗-HIVIgG和IgA抗体水平。结果D、M和R疫苗均可诱导小鼠产生广泛的免疫应答,ELISPOT法可检出脾细胞分泌IFNγ、IL-2、IL-4和IL-6;胞内因子染色可检出IFNγ、IL-2和IL-4的分泌;D和M组脾淋巴细胞增殖试验阳性数高于R组;D、M和R组血清抗-HIVIgG抗体分别有0/5、1/5和5/5小鼠阳转,各组均未检出血清抗-HIVIgA。结论已初步建立了HIV疫苗小鼠免疫原性检测方法。

Objective To preliminarily develop a method for determination of immunogenicity of HIV vaccine in mice and evaluate the immunogenicity. Methods BALB / c mice were immunized with one kind of HIV DNA vaccine（D）and two kinds of recombinant vaccinia virus-based HIV vaccines（M and R）respectively, and their sera and splenic lymphocytes were separated 6 weeks later. The secretions of cytokines in splenic lymphocytes were determined by ELISOPT and intracellular cytokine staining, and the proliferation of lymphocytes by MTS method. The anit-HIV IgG and IgA levels in sera were determined by ELISA. Results All the vaccines induced a broad of immune responses in mice. ELISPOT proved the secretions of IFNγ, IL-2, IL-4 and IL-6, and intracellular cytokine staining proved the secretions of IFNγ, IL-2 and IL-4 in splenic lymphocytes. The numbers of splenic lymphocytes positive in proliferation test of mice immunized with vaccines D and M were larger than that with vaccine R. Anti-HIV IgG were detected in 0 / 5, 1 / 5 and 5 / 5 of mice immunized with vaccines D, M and R respectively, while no anti-HIV IgA were detected. Conclusion A method for determination of immunogenicity of HIV vaccine in mice was preliminarily developed.