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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 establish an immunogenicity test for HIV vaccine in mice and to evaluate the immunogenicity of HIV vaccine. Methods One HIV DNA vaccine (D) and two recombinant vaccinia carrier HIV vaccines (M and R) were selected and BALB / c mice were immunized respectively. Serum and splenic lymphocytes were isolated after 6 weeks. ELISOPT and intracellular factor staining Cytokine secretion, MTS assay lymphocyte proliferation response, ELISA detection of serum anti-HIV IgG and IgA antibody levels. Results D, M and R vaccines could induce a broad immune response in mice. IL-2, IL-4 and IL-6 secreted by spleen cells were detected by ELISPOT. IFNγ, IL -2 and IL-4 secretion; the positive numbers of splenic lymphocyte proliferation test in group D and M were higher than those in group R; the serum anti-HIVIgG antibodies in groups D, M and R were 0/5, 1/5 and 5/5 respectively Rats were positive in rotation, serum anti-HIVIVA was not detected in all groups. Conclusion The immunogenicity of HIV vaccine was initially established.