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目的 研究乙型肝炎表面抗原(HBsAg)不同突变体对细胞免疫的影响。方法 将野生型和变异型HBsAg基因重组质粒NS2 Swt、NS2 S12 6、NS2 S133、NS2 S14 1、NS2 S14 5分别转染CHO细胞,72h收获细胞上清。采用ELISA法检测各组细胞上清preS2 蛋白的表达量。将这些细胞上清刺激PHA活化的人淋巴细胞,MTS法检测不同变异株抗原对淋巴细胞增殖活性以及淋巴细胞分泌IFN γ、IL 10和IL 2的影响。结果 变异和野毒株(wt)各组细胞上清preS2 蛋白的表达量基本一致。变异和wt重组HBsAg刺激T细胞后,其上清MTS显色后的A4 90 值均高于空白组和pCI neo组,说明HBsAg中的蛋白可以促进T细胞增殖;T12 6S氨基酸变异HBsAg能够刺激IFN γ分泌增加;M133T氨基酸变异刺激IL 10分泌增加。结论 这4种乙型肝炎病毒(HBV)变异株感染人体后,机体对它们的细胞免疫反应可能不会有明显的增强或减弱,但也不能忽视T12 6S和M133T变异抗原改变了某些细胞因子的表达,可能对机体细胞免疫造成的潜在影响。
Objective To study the effect of different mutants of hepatitis B surface antigen (HBsAg) on cellular immunity. Methods The recombinant plasmids of NS2 Swt, NS2 S12 6, NS2 S133, NS2 S14 1 and NS2 S14 5 of wild type and variant HBsAg gene were transfected into CHO cells, and the supernatant of cells were harvested at 72 hours. The expression of preS2 protein in supernatant of each group was detected by ELISA. The supernatant of these cells stimulated PHA-activated human lymphocytes, MTS assay of different mutant antigens on lymphocyte proliferation activity and lymphocyte secretion of IFN γ, IL 10 and IL 2. Results The expression of preS2 protein in the supernatant of the mutated and wild-type strain (wt) groups was basically the same. After stimulation with T cells by mutated and recombinant HBsAg, the A490 values of MTS after supernatants were higher than that of blank group and pCI neo group, indicating that HBsAg protein can promote T cell proliferation; T12 6S amino acid mutation HBsAg can stimulate IFN γ secretion increased; M133T amino acid mutation stimulates IL 10 secretion increased. CONCLUSION: The four hepatitis B virus (HBV) mutants may not significantly enhance or diminish their cellular immune response after being infected in humans. However, we can not ignore the fact that the T12 6S and M133T variant antigens change certain cytokines The potential impact on cellular immunity of the body.