儿童新甲型流感疫苗远期特异性CD8＋记忆T细胞的表型特征

摘要

目的：探讨儿童新甲型（ H1 N1）流感疫苗接种后远期外周血疫苗特异性CD8＋记忆T细胞的特征。方法：选择31例于2009年12月～2010年1月接种甲型H1N1流感疫苗的3～6岁儿童，抽取外周静脉血5 ml，分离淋巴细胞加入甲型H1 N1流感疫苗进行培养，并设对照组，用流式细胞仪检测细胞表面分子的表达，同时以CCK－8法检测细胞增殖，计算细胞增殖指数。结果：外周血PBMC中CD8＋T细胞在实验组和对照组分别是13．41％与9．41％，P＞0．05。 CD8＋初始T细胞在两组的比例均高达80％上；CD8＋记忆T细胞在两组占17％～19％；记忆T细胞分中央型和效应型记忆T细胞，两个亚群检测结果：CCR7和CD62L单阳性记忆T细胞亚群在实验组占比明显低于对照组的占比，P＜0．05。应用CCK－8法检测疫苗刺激后抗原特异性增殖，检测结果见细胞增殖指数大于0．8的仅为51．16％，大于1．0在本研究中没有。结论：31例儿童外周血PBMC中CD8＋T细胞占比少，初始T细胞CD45 R＋均较高，具有抗原再刺激后再应答的基础；甲型H1 N1流感疫苗可以诱导抗原特异性记忆CD8＋T细胞的产生，但数量不多，其中大多数为中央型记忆CD8＋T细胞；细胞增殖指数结果表明疫苗特异性细胞增殖不佳；提示对该疫苗的远期疫苗特异性CD8＋T细胞免疫记忆不理想。%Objective:To elucidate the characterization of CD8+T cell in H1N1 influenza vaccine for children.Methods:PBMCs were isolated from 31 children aged from 3 to 6 years old who had accepted H 1N1 influenza vaccine during December 2009 to January 2010.The lymphocytes were joined with the H 1N1 influenza vaccine as experimental group and cultured .The experiment set without vaccine group as control group .At last we detected the surface molecules by FCM .The CCK-8 assay was added to detecting cellular proliferation and cellular proliferation index were detected by CCK-8.Results: CD8+T cells of PBMC in the two groups were 13.41%and 9.41%,P>0.05.CD8+CD45RAA+naive T cells in the two groups were up to more than 80%,P>0.05.The proportion of CD8+CD45ROA+memory T cells in two groups were up to 17%-19%,P>0.05.Two subsets of CD8+CD45ROA+memory T cells :CCR7+and CD62L+single positive memory T cell subsets in the experimental group were significantly lower than that of the control group,P<0.05.The CCK-8 assay was added to detect cellular proliferation .Only 51.16% of which cellular proliferation index was greater than 0.8,with none was greater than 1 in this study.Conclusion:This study showed that the CD4+T cells were low-level,naive T cells (CD8+CD45RAA+)were higher,with antigen stimulation and response.H1N1 vaccination specific memory T cells were few in number , specific memory T cell subsets were diversity , control memory cells were the main phenotypic characteristics .Cellular proliferation index showed that the proliferation of specific CD 8+T cells vaccine was poor .