成人接种乙型肝炎疫苗后细胞免疫应答的动态变化

Study on the kinesis of cellular immunity in adults vaccinated with recombinant hepatitis B vaccine

目的探讨人体接种乙型肝炎（乙肝）疫苗后细胞免疫的动态变化,了解疫苗介导的细胞免疫应答与体液免疫应答的关联。方法8例成人按0、1、2月程序接种乙肝疫苗,于首针免疫后3、8、21、34和65 d采静脉血,分离外周血单个核细胞（PBMC）,对PBMC进行细胞分选,获得纯度95%以上的CIM^＋和CD8^＋ T淋巴细胞,应用酶联免疫斑点法（ELISPOT）测定PBMC、CD4^＋和CD8^＋ T淋巴细胞体外刺激后所产生的细胞因子IFN-γ、IL-2和IL-4的斑点数（SFC）,并收集乙肝疫苗免疫前（0 d）和免疫后3、8、21、30、34、60、65和150 d血样,测定乙肝表面抗体（抗-HBs）。同时设立阴、阳性对照各2例。结果人接种乙肝疫苗后,CD4^＋、CD8^＋ T淋巴细胞产生不同细胞因子趋势和强度不同： IFN-γ（由CD8^＋、CIM^＋ T淋巴细胞产生）出现较早,且较为稳定,而CD4^＋ T淋巴细胞产生的IL-2和IL-4出现较晚,但其升高与第2、3针乙肝疫苗免疫有关;免疫后IL-4阳转率与抗-HBs阳转率呈显著正相关,IL-2、IL-4的SFC与抗-HBs滴度也呈显著正相关。结论成人接种乙肝疫苗后可早期检测出以IFN-γ为主的细胞免疫,IL-4和IL-2阳转与抗-HBs升高有关。

Objective To evaluate the kinesis of cellular immunity in adults who were vaccinated with yeast recombinant hepatitis B（rHB） vaccine and the correlation between cellular and humoral immune responses induced by the vaccine. Methods Eight adults were vaccinated with rHB vaccine according to 0, 1,2 month schedule. The peripheral blood mononuclear cells（PBMCs） were collected at the 3, 8, 21, 34 and 65 days after the first dose. The high purity of CD4^＋ and CD8^＋ T cells obtained by sorting from PBMCs were re.stimulated with recombinant hepatitis B surface antigens（rHBsAg） or peptides. The spot forming cell（SFC） of IFN-γ, IL-2 and IL-4 of CD4^＋ and CD8^＋ T cells were detected by enzyme-linked immunospot（ELISPOT）. Results The characteristics of IFN-γ,IL-2 and IL-4 of CD4^＋ and CD8^＋ T cells appeared different after immunization with rHB vaccine. IFN-γ of CD8^＋ and CD4^＋ T cells could be detected early with stable SFC, while the IL-2 and IL-4 of CD4^＋ T cells appeared late but increased after the second and third dose of vaccination. The positive rate of IL-4 of CD4^＋ T cells were significantly correlated with the positive rate of anti-FIBs, while the SFCs of IL-4 and IL-2 of CD4^＋ T cells were also significantly related to the titers of anti-HBs. Conclusion IFN-γ could be detected early after rHB vaccination in adults, and the positive rates of IL-4 and IL-2 were correlated with that of anti-HBs.