HBV相关肝病患者外周血NK细胞PD-1、Tim-3表达及意义

Expression and Meaning of PD-1,Tim-3 on NK in HBV Related Liver Disease

目的了解HBV相关肝病患者外周血NK细胞表面PD-1、Tim-3的表达情况,探讨PD-1和Tim-3的表达与血清HBV-DNA载量的相关情况。方法选择HBV感染相关肝病患者74例,其中CHB患者30例,LC患者24例,HCC患者20例,健康对照18例。应用流式细胞术检测NK细胞表面PD-1、Tim-3表达,荧光定量PCR检测HBV-DNA量,ELISA法检测细胞因子分泌水平。统计各组患者PD-1和Tim-3的表达率,比较各组差异,分析其与病情的相关性。结果 CHB、LC、HCC组外周血NK细胞表面PD-1和Tim-3的表达率分别为34.46%±7.31%、59.09%±12.35%、61.49%±15.26%和19.18%±6.05%、28.64%±11.20%、31.24%±11.85%,均高于对照组(P<0.01),LC组和HCC组明显高于CHB组(P<0.01),LC组和HCC组比较无统计学差异。在CHB组,PD-1的表达率与HBV-DNA呈负相关关系(R=-0.437,P=0.033)。CHB、LC、HCC各组血清中细胞因子TNF-α和IFN-γ水平分别为0.66±0.63、0.75±0.66、0.85±0.71和1.76±0.88、1.83±1.23、1.60±0.53,均低于健康对照组(P<0.01)。结论 HBV相关肝病患者NK细胞表面PD-1、Tim-3表达均增加,且与病情严重程度有关,增加表达的PD-1与血清HBV-DNA载量存在负相关性,HBV肝病各组血清TNF-α、IFN-γ分泌减少。

Objective To quantitative analysis the expression variation of PD-1, Tim-3 on HBV-related liver disease patients peripheral blood NK cell. To investigate the correlation of serum HBV-DNA loads and the expression of PD-1, Tim-3. Methods All of 92 subjects including 30 CHB, 24 LC, 20 HCC and 18 healthy adults were enrolled in present study. The expression of PD-1 and Tim-3 were detected by flow cytometry. The quantity of HBV-DNA were obtained by fluorescence quantitative PCR, and the concentrations of cytokines were assayed by enzyme linked immunosorbent assay（ELISA）.The expression of PD-1, Tim-3 in each group, was Calculated and compared the differences, analyzed its correlation with disease progression. Results In group of CHB, LC, HCC, expression of PD-1 and Tim-3 on peripheral blood NK cell were 34.46%±7.31%, 59.09%±12.35%, 61.49%±15.26% and 19.18%±6.05%,28.64%±11.20%,31.24%±11.85% respectively, which were higher than control group （P〈0.01）. Group of LC and HCC were higher than CHB group （P〈0.01）, but no significant difference were found between LC and HCC group. In the group of CHB, the expression of PD-1 and HBV-DNA loads was negatively correlated（R=-0.437, P=0.033）. Compared with the group of control, the concentration of TNF-α and IFN-γ decreased in the group of CHB, LC, HCC 0.66±0.63, 0.75±0.66, 0.85±0.71 and 1.76±0.88, 1.83±1.23, 1.60±0.53, （P〈0.01）. Conclusion The expression of PD-1, Tim-3 on NK cell were increased in HBV-related liver disease patients, and the increased expression of PD-1 was negatively correlated with serum HBV-DNA loads. Concentration of TNF-α,IFN-γ were decreased in HBV related liver disease patients.