Li Jin-cheng, Tan De-ming, Liu Hong-bo, Li Ke-cheng
Department of Infectious Diseases, Central South University, Changsha 410008, China.
Zhonghua Gan Zang Bing Za Zhi. 2010 Oct;18(10):726-30. doi: 10.3760/cma.j.issn.1007-3418.2010.10.002.
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.
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 peripheral-blood 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) gamma.
(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 value less than 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 IFNgamma 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 value less than 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.
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 IFNgamma; There was no correlation between the titer of anti-HBs and the expressions of IFNgamma and CD4+CD25+ Foxp3.
通过检测不同反应组中调节性T细胞(Tregs)的表达、Foxp3 mRNA以及乙肝疫苗接种后细胞因子的分泌情况,探讨免疫反应与Tregs及细胞因子之间的内在联系。
采集不同反应组的血样。采用实时荧光定量PCR检测外周血单个核细胞中Foxp3 mRNA的表达;用流式细胞术检测外周血单个核细胞表面标志物CD4和CD25;用ELISA法检测外周血单个核细胞在植物血凝素(PHA)、乙肝表面抗原(HBsAg)刺激下产生白细胞介素-4(IL -4)、白细胞介素-12(IL-12)、白细胞介素-18及γ干扰素(IFN)的水平。
(1)PHA和HBsAg刺激前后,反应组和无反应组的Foxp3表达均较高。差异有统计学意义(P值小于0.05);(2)外周血中,反应组CD4+T细胞中CD4+CD25+ Treg的百分比(0.59%±0.46%)明显低于对照组(1.30%±1.44%);(3)各组PHA和HbsAg刺激外周血单个核细胞后,无反应组γ干扰素浓度[(11.00±9.03)IU/ml]明显低于反应组[(38.88±28.16)IU/ml],差异有统计学意义(P值小于0.01);(4)PHA或HBsAg刺激外周血单个核细胞后,IL-18、IL-4和IL-12浓度无显著差异。
CD4+CD25+Foxp3+ Treg细胞在一定程度上可能参与乙肝疫苗接种免疫反应的负调控。乙肝疫苗接种免疫无反应可能与γ干扰素分泌不足有关;抗-HBs滴度与γ干扰素及CD4+CD25+ Foxp3的表达无相关性。