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目的利用AE亚型的膜蛋白,研发抗HI V-1 AE重组型的疫苗并进行免疫原性评价。方法构建表达中国流行株97CNGX2F(HI V-1 AE2F)包膜蛋白gp140的重组痘苗病毒rVVT140AE。使用表达gp140的DNA疫苗和rVVT140AE用初免-加强方式免疫小鼠。免疫结束后第2周处死小鼠,分别检测特异性抗体滴度和特异性T细胞反应。结果本疫苗所活化的特异性抗体滴度在3 200和51 200之间,几何平均值为11 143;总T细胞免疫反应强度为(1 918±442)SFCs/106脾细胞,其中针对env1、env2、env3、env4肽池的免疫反应强度分别为(1 280±330)SFCs/106脾细胞,(66±16)SFCs/106脾细胞,(163±34)SFCs/106脾细胞和(409±96)SFCs/106脾细胞,均显著高于空载体对照组(<10 SFCs/106脾细胞)。结论本实验构建的HI V-1 AE2F株包膜蛋白gp140重组痘苗病毒疫苗可作为针对我国HI V-1流行株的候选疫苗免疫原。
OBJECTIVE To develop a membrane antigen of AE subtype and develop an anti-HI V-1 AE recombinant vaccine for immunogenicity evaluation. Methods Recombinant vaccinia virus rVVT140AE, which expressed the envelope protein gp140 of Chinese popular strain 97CNGX2F (HI V-1 AE2F), was constructed. Mice were immunized with prime-boost using a DNA vaccine expressing gp140 and rVVT140AE. Mice were sacrificed at the second week after immunization, and specific antibody titers and specific T cell responses were tested. Results The titer of specific antibody activated by this vaccine was between 3 200 and 51 200 with a geometric mean of 11 143. The total T cell immunoreactivity was (1 918 ± 442) SFCs / 106 splenocytes, of which env1, The immunoreactivity of env2, env3 and env4 peptide pools were (1 280 ± 330) SFCs / 106 splenocytes, (66 ± 16) SFCs / 106 splenocytes, (163 ± 34) SFCs / 106 splenocytes and 96) SFCs / 106 splenocytes were significantly higher than the empty vector control group (<10 SFCs / 106 splenocytes). Conclusion The HI V-1 AE2F strain envelope protein gp140 recombinant vaccinia virus vaccine constructed in this study can be used as a candidate vaccine immunogen for HI V-1 epidemic strains in our country.