慢性HBV感染者HBV特异性CD8＋ T细胞Tim-3和PD-1的表达水平及其与IFN-γ产生的相关性研究

目的：研究免疫球蛋白黏蛋白分子-3（Tim-3）和程序性死亡受体-1分子（PD-1）在慢性乙型肝炎病毒（HBV）感染患者外周血HBV特异性CD8＋ T细胞表面的表达模式，了解其与γ-干扰素（IFN-γ）产生的关系，并探讨其临床意义。方法采用流式细胞术检测主要组织相容性复合体-2（HLA-A2）阳性的78例临床类型不同的慢性HBV感染患者HBV特异性CD8＋ T细胞表面分子Tim-3和PD-1的表达，ELISA方法检测外周血单个核细胞（PBMC）培养上清液中IFN-γ的水平。结果慢性HBV感染者Tim-3＋/PD-1＋HBV特异性CD8＋ T细胞比例占总的HBV特异性CD8＋ T细胞的58%，Tim-3-/PD-1＋细胞比例为24%，Tim-3-/PD-1-比例16%，Tim-3＋/PD-1-比例最低为2%。临床病情越严重的临床类型中，Tim-3＋/PD-1＋HBV特异性CD8＋ T细胞比例越高，慢性乙型肝炎轻中度组为（52.05±18.68）%，重度肝炎组为（59.66±19.25）%，重型肝炎组最高为（68.72±17.21）%；各组与非活动性携带者组比较，P值分别为0.007、0.009、0.000。重型组与轻中度组比较，P =0.018。Tim-3＋/PD-1＋在HBV特异性CD8＋ T细胞的表达与细胞培养上清液IFN-γ的水平呈负相关性（r =-0.466，P ＜0.001）。结论在HBV特异性CD8＋ T细胞中，Tim-3和PD-1共同表达是其主要表达模式，Tim-3和PD-1的高表达可能负性调控IFN-γ的产生，从而影响慢性HBV感染的疾病进展和结局。

The expression of Tim-3 and PD-1 on HBV specific CD8 T cells in chronic HBV infection patients and its correlation with the production of interferon gamma

Objective To study the patterns of expression of T-cell immunoglobulin and mucin domain-containing molecule 3 （Tim-3） and programmed death1 （PD-1） on HBV specific CD8＋ T cells in chronic hepatitis B virus infection patients （CHB） . Ivestigate the correlation between Tim-3 and PD-1 and production of interferon gamma （IFN-γ） and its clinical meanings. Methods The expression of Tim-3 and PD-1 on HBV specific CD8＋ T cells in 78 major histocompatibility complex-2 （HLA-A2） positive HBV patients with different clinical types were detected by flow cytometry. The level of IFN-γ in the supernatant of culture of peripheral blood mononuclear cell （PBMC） were detected by double antibody sandwich ELISA. Results In chronic HBV infection patients, 58% of HBV-specific CD8＋ T cells are Tim-3＋/PD-1＋ cells, 24%are Tim-3-/PD-1＋ cells. The ratio of Tim-3-/PD-1- cells was 16% and the Tim-3＋/PD-1- cells are the lowest with 2%. The proportion of Tim-3＋/PD-1＋ HBV-specific CD8＋ T cells increased consistent with the clinical severity. The ratio of Tim-3＋/PD-1＋ HBV-specific CD8＋ T cells is （33.93 ± 10.80）% in the inactive HBsAg carriers. The ratio of Tim-3＋/PD-1＋ cells in mild and moderate chronic hepatitis patients are （52.05 ± 18.68）%, （59.66 ± 19.25）% are Tim-3＋/PD-1＋ cells in severe hepatitis, and （68.72 ± 17.21）% are Tim-3＋/PD-1＋ cells in hepatic failure, which is the highest. Compared with inactive HBsAg carriers, P = 0.007, 0.009, 0.000. Compare hepatic failure and mild and moderate chronic hepatitis, P = 0.018. The percentage of Tim-3＋/PD-1＋ HBV-specific CD8＋ T cells negatively correlate with the level of IFN-γ in culture supernatant. Conclusions sCo-expression of Tim-3 and PD-1 on HBV-specific CD8＋ T cells is the main pattern in HBV patients. High expression of Tim-3 and PD-1 may negatively control the production of IFN-γ, which may influence the outcomes and disease progression in chronic HBV infection.