调节性T淋巴细胞对成人乙型肝炎疫苗接种免疫应答的影响  

The roles of CD4＋CD25＋ T regulatory cells and Foxp3 mRNA expression in the subjects with and with out responses to hepatitis B virus vaccination

作　　者：李金成[1,2] 谭德明[1] 刘洪波[1] 李科成[3] 

机构地区：[1]中南大学湘雅医院感染科,长沙410008 [2]邵阳医学高等专科学校内科学教研室 [3]珠海市妇幼保健院检验科

出　　处：《中华肝脏病杂志》2010年第10期726-730,共5页Chinese Journal of Hepatology

基　　金：基金项目：湖南省科技厅项目（2007FJ3092）;湖南省教育厅项目（07C065）;湖南省卫生厅项目（B2007194）

摘　　要：目的 通过对乙型肝炎疫苗接种后不同免疫应答人群调节性T淋巴细胞（Treg细胞）Foxp3 mRNA的表达和细胞因子分泌的检测,探讨乙型肝炎疫苗接种后免疫应答与免疫调节细胞和细胞因子之间的内在联系.方法 采集不同反应人群全血,实时荧光定量PCR检测人外周血单个核细胞Foxp3 mRNA的表达;流式细胞术对外周血单核细胞的表面标志物CD4、CD25进行检测;酶联免疫法检测外周血单个核细胞植物血凝素（PHA）和HBsAg刺激后白细胞介素（IL）-4、IL-12、IL-18、干扰素（IFN）γ的产生水平.根据不同资料采用方差分析、q检验或相关分析进行统计学处理.结果 （1）PHA和HBsAg刺激前后,无应答组Foxp3的表达均高于应答组和对照组,差异有统计学意义（P＜0.05）.（2）应答组外周血CD4＋CD25＋Treg细胞占CD4＋T淋巴细胞的百分比（0.59%±0.46%）明显低于对照组（1.30%±1.44%）,差异有统计学意义（F=2.990,P＜0.01）.（3）各组外周血单个核细胞经PHA和HBsAg刺激后,无应答组的IFN γ浓度[（11.00±9.03）U/ml]明显低于应答组[（38.88±28.16）U/ml],差异有统计学意义（P＜0.01）.（4）各组外周血单个核细胞经PHA和HBsAg刺激后,IL-18、IL-4、IL-12的浓度差异均无统计学意义.结论 CD4＋CD25＋Foxp3＋Treg细胞在一定程度上参与了乙型肝炎疫苗接种应答的负性调控;乙型肝炎疫苗接种后无应答与IFN γ分泌不足有关;抗-HBs滴度水平与IFN γ和CD4＋CD25＋Foxp3表达无相关性.Objective To investigate the internal links between immune responses and Tregs and cytokine by the expression of T regulatory cells （Tregs）, Foxp3 mRNA of different response groups and the detection of cytokine secretion after hepatitis B vaccination. Methods Blood samples were collected in different response groups. Real-time fluorescence quantitative PCR was used to detect the expression of Foxp3 mRNA of peripheral blood mononuclear cells; The surface markers CD4 and CD25 in peripheralblood mononuclear cells were determined by flow cytometry; ELISA tests were used to detect the production level of phytohemagglutinin （PHA） in peripheral blood mononuclear cells, IL -4, IL-12, IL-18 stimulated by HBsAg and （IFN） γ. Results （1） Foxp3 expressions in response group and non-response group were higher before or after PHA and HBsAg were stimulated. Differences were statistically significant （P 〈 0.05）; （2） In peripheral blood, the percentage of CD4＋CD25＋ Treg of CD4＋ T cells in response group （0.59% ± 0.46%） was obviously lower than those in control group （1.30% ± 1.44%）; （3） Peripheral blood mononuclear cells stimulated by PHA and HbsAg in each group, the concentration of IFN γ in non-response group [（11.00 ±9.03） IU/ml] was markedly lower than those in response group [（38.88 ± 28.16） IU/ml],and differences were statistically significant （P 〈 0.01）; （4） In PHA- or HBsAg-stimulated peripheral-blood mononuclear cells,the concentrations of IL-18, IL-4 and IL-12 had no significant difference. Conclusions To some extent,CD4＋CD25＋Foxp3＋ Treg cells may be involved in negative regulation of the immune responses to HBV vaccination.Immune non-response to HBV vaccination may be connected to insufficient secretion of IFN γ; There was no correlation between the titer of anti-HBs and the expressions of IFN γ and CD4＋CD25＋ Foxp3.

关 键 词：肝炎疫苗 乙型 T淋巴细胞 调节性 抗原 CD4 抗原 CD25 FOXP3 

分 类 号：R686[医药卫生—骨科学]