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目的检测慢性乙型病毒性肝炎患者外周血中iNKT细胞比例、细胞因子分泌能力以及其表面程序性死亡因子1(programmed death 1,PD-1)的表达情况。方法采集慢性乙型肝炎患者外周血,利用CD3、TCR Vα24、TCR Vβ11、CD279单克隆抗体标记后经流式细胞术直接检测iNKT细胞比例及其PD-1表达比例;采用PMA+Ionomycin体外活化iNKT细胞后流式细胞术检测其胞内IFN-γ分泌情况。结果慢性乙型肝炎患者外周血iNKT细胞比例[(0.09±0.04)%]与健康对照者[(0.11±0.07)%]相比无明显差异,但体外活化后胞内IFN-γ分泌量减少[(24.64±7.71)%vs(42.35±11.60)%],且iNKT细胞PD-1表达升高[(36.7±9.7)%vs(16.2±5.4)%]。结论慢性乙型肝炎患者外周血iNKT细胞功能的下降可能与其表面负性刺激分子PD-1表达上调密切相关。
Objective To detect the proportion of iNKT cells, the secretion of cytokines and the expression of programmed death 1 (PD-1) in peripheral blood of patients with chronic hepatitis B virus. Methods Peripheral blood of patients with chronic hepatitis B were collected and directly labeled with CD3, TCR Vα24, TCR Vβ11 and CD279 monoclonal antibodies to detect the proportion of iNKT cells and their PD-1 expression ratio by flow cytometry. In vitro activation of iNKT by PMA + Ionomycin Cell flow cytometry was used to detect the intracellular IFN-γ secretion. Results The percentage of iNKT cells in peripheral blood of patients with chronic hepatitis B [(0.09 ± 0.04)%] was not significantly different from that in healthy controls [(0.11 ± 0.07)%], but the amount of intracellular IFN- (24.64 ± 7.71)% vs (42.35 ± 11.60)%], and the expression of PD-1 was increased in iNKT cells [(36.7 ± 9.7) vs (16.2 ± 5.4)%]. Conclusion The decreased function of iNKT in peripheral blood of patients with chronic hepatitis B may be closely related to the up-regulation of PD-1 expression on the surface.