HCV感染者直接抗病毒药物治疗前后CD8^+T淋巴细胞衰老和功能相关指标的变化

Changes in senescence-and function-related parameters of CD8^+T cells after direct-acting antiviral treatment in patients with hepatitis C virus infection

目的观察HCV感染者DAA治疗前后CD8^+T淋巴细胞衰老、功能相关指标变化,并探讨其临床意义。方法选取2017年1月-2018年12月于空军军医大学第二附属医院就诊的HCV感染者26例,患者接受索磷布韦联合达卡他韦片治疗。并纳入治愈者22例,健康对照者20例。采用流式细胞仪检测CD8+T淋巴细胞上SIRT1、CD57、PD-1、Tim-3等相关分子表达,并采用RT-PCR方法检测p21、p53表达水平,Luminex液相悬浮芯片检测样本外周血衰老相关分泌表型。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 SIRT1、PD-1、Tim-3在3组间表达差异均有统计学意义(F值分别为6. 712、4. 202、4. 575,P值均<0. 05)。与健康对照组相比,HCV组CD8+T细胞上SIRT1、PD-1、Tim-3表达水平明显上升(P值均<0. 05),HCV治愈组Tim-3表达水平明显上升(P <0. 05),HCV治愈组SIRT1、PD-1表达水平无明显变化(P值均> 0. 05)。p53、p21在3组间表达差异有统计学意义(F值分别为11. 144、6. 594,P值均<0. 05)。与健康对照组相比,HCV治愈组和HCV组p53表达水平明显下降(P值均<0. 001),p21表达水平明显下降(P值均<0. 05),HCV组和HCV治愈组比较差异均无统计学意义(P值均> 0. 05)。IL-6、TNFα在3组间差异均有统计学意义(F值分别为3. 920、6. 337,P值均<0. 05),与健康对照组相比,HCV组外周血的IL-6和TNFα表达水平明显升高(P值均<0. 05),HCV治愈组IL-6和TNFα表达水平无明显变化(P值均> 0. 05),与HCV组相比,HCV治愈组TNFα表达水平明显下降(P=0. 007)。结论 HCV感染者CD8+T淋巴细胞衰老,经DAA治疗后衰老缓解,CD8+T淋巴细胞功能部分恢复。

Objective To investigate the changes in the senescence-and function-related parameters of CD8^+T cells after direct-acting antiviral( DAA) treatment and their clinical significance in patients with hepatitis C virus( HCV) infection. Methods A total of 26 patients with HCV infection who attended the Second Affiliated Hospital of Air Force Medical University from January 2017 to December 2018 were enrolled,and all patients were given sofosbuvir and daclatasvir tablets. A total of 22 patients who were cured and 20 healthy controls were enrolled. Flow cytometry was used to measure the expression of silent information regulator 1( SIRT1),CD57,programmed death-1( PD-1),and Tim-3 on CD8^+T cells;RT-PCR was used to measure the expression of p21 and p53;Luminex liquid suspension chip was used to observe senescence-associated secretory phenotype in peripheral blood. A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups. Results There were significant differences in the expression of SIRT1,PD-1,and Tim-3 between the three groups( F = 6. 712,4. 202,and 4. 575,all P < 0. 05). Compared with the healthy control group,the HCV group had significant increases in the expression of SIRT1,PD-1,and Tim-3 on CD8^+T cells( all P < 0. 05),and the HCV cured group had a significant increase in the expression of Tim-3( P< 0. 05),with no significant changes in SIRT1 and PD-1( both P > 0. 05). There were significant differences in the expression of p53 and p21 between the three groups( F = 11. 144 and 6. 594,both P < 0. 05). Compared with the healthy control group,the HCV cured group and the HCV group had significant reductions in the expression of p53( both P < 0. 001) and p21( both P < 0. 05),and there were no significant differences between the HCV group and the HCV cured group( both P > 0. 05). There were significant differences in interleukin-6 ( IL-6) and tumor necrosis factor α( TNFα) between the three groups( F = 3. 920 and 6. 337,both P < 0. 05),and compared with the healthy control group,the HCV group had significant increases in the levels of IL-6 and TNFα in peripheral blood( both P < 0. 05). There were no significant changes in IL-6 and TNFα in the HCV cured group( both P > 0. 05),and compared with the HCV group,the HCV cured group had a significant reduction in the level of TNFα( P = 0. 007). Conclusion Senescence of CD8^+T cells is observed in patients with HCV infection and is alleviated after DAA treatment,with partial recovery of the function of CD8^+T cells.