摘要
目的观察聚乙二醇干扰素（Peg-IFN）α-2a联合利巴韦林治疗慢性丙型肝炎初治患者抗病毒治疗48周及停药后随访至72周外周血单个核细胞（PBMC）表面程序性死亡受体（PD）-1、Toll样受体（TLR）3、TLR4表达率的变化，探讨PBMC表面PD-1、TLR3、TLR4是否参与了宿主抗HCV固有免疫和获得性免疫过程，及其与持续病毒学应答的关系。方法收集慢性丙型肝炎初治患者23例及正常人10名，慢性丙型肝炎患者均接受48周Peg—IFNα-2a（180μg／周）联合利巴韦林口服（15μ·kg-1·d。）抗病毒治疗。分别在0、4、12、24、48、72周时，检测患者血清中ALT、HCVRNA变化；分离患者PBMC，流式细胞术检测不同时间点患者和正常人PBMC表面PD-1、TLR3、TLR4的表达率，分析持续病毒学应答（SVR）与非SVR患者上述指标的变化和差异。多组资料之间的比较采用onewayANOVA模块进行t检验；两组之间比较采用独立样本f检验。结果丙型肝炎患者基线时与正常人PBMC表面PD-1、TLR4、PDl／TLR4表达率分别为：45．44％±7．32％、57．74％士15．45％、35．42％±7．97％与16．82％±4．13％、21．09％±2．89％、14．12％±2．89％，丙型肝炎患者基线水平较正常人明显升高（t值分别为4．675，5．148，2．549，尸值均〈0．05）。正常人TLR3及PDl／TLR3表达率较丙型肝炎患者基线水平稍低，但差异无统计学意义（P〉0．05）。在Peg—IFNα-2a联合利巴韦林治疗期间，慢性丙型肝炎患者PBMC上PD-1及TLR4表达率逐渐下降，TLR3表达率逐渐升高；且SVR患者PBMC上PD-1和TLR3表达率的变化与非SVR患者的差异明显（P〈0．05）。结论慢性丙型肝炎患者PBMC上PD-1、TLR4、PDl／TLR4高表达，初治患者用Peg-IFNα-2a治疗可以使这些指标表达下降，TLR3的表达率升高；PBMC表面PD-1和TLR3的表达可能与患者出现SVR密切相关。
Objective To investigate the dynamic changes in expression of programmed death （PD）-I, Toll-like receptor （TLR）3, and TLR4 on the surface of peripheral blood mononuclear cells （PBMCs） in patients with chronic hepatitis C （CHC） that occur in response to pegylated-interferon alpha-2a （peg-IFNct- 2a） vlus ribavirin （RBV） combination theraov, and to analvze the relation to achievement of sustained virological response （SVR）. Methods Twenty-three CHC patients and 10 healthy controls were enrolled in the study. All CHC patients underwent 48 weeks of combination therapy with peg-IFNct-2a （180 ~tg, subcutaneous injection, once weekly） plus RBV （15 ~tg/kg, oral, once daily）. Total PBMCs were isolated from both groups （CHC patients at treatment week 0, 12, 24, and 48 and post-treatment week 24; controls at enrollment） and subjected to flow cytometric analysis of PD-1, TLR3, and TLR4 surface expression. In addition, serum levels of alanine aminotransferase （ALT） and hepatitis C virus （HCV） RNA levels were analyzed by enzymatic assay and the AmpliPrep/COBAS （Roche） nucleic acid amplification test, respectively. SVR was defined as undetectable levels of HCV RNA at post-treatment week 24. Intergroup differences were assessed by one- way ANOVA. Results The expression ratios of PD-1, TLR4 and PD-I： TLR4 on PBMCs were significantly higher in CHC patients before therapy than in the healthy controls （45.20 -4- 7.12% vs. 16.82 -4- 4.13%, 58.45 ＋ 15.13% vs. 21.09 ~ 2.89%, and 35.54 q- 7.69% vs. 14.12 q- 2.89%; allP 〈 0.05）. In contrast, the expression ratios of TLR3 and PD-1 ：TLR3 were slightly, but not significantly, higher in CHC patients before therapy than in the healthy controls （P 〉 0.05）. During the course of peg-IFNct-2a plus RBV combination therapy, the expression ratios of PD-1 and TLR4on PBMCs showed a decreasing trend, while TLR3 expression showed an increasing trend. Furthermore, CHB patients who achieved SVR at post-treatment week 24 had a significantly different expression ratio of PD-1 and TLR3 than those who did not achieve SVR （_P 〈 0.05）. Conclusion Surface expression of PD-1, TLR4, and PD-I：TLR4 is up-regulated in the total PBMCs of CHC patients. Peg- IFNct-2a plus RBV treatment-induced suppression of HCV replication results in a significant reduction in PD-1 and TLR4 expression on the surface of PBMCs, but a remarkably elevated level of TLR3 expression. The dynamic change in PD-1 and TLR3 expression on PBMCs that occurs during antiviral therapy may be related to achievement of SVR.
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2013年第3期196-201,共6页
Chinese Journal of Hepatology
基金
国家自然科学基金（30901269）
国家“十一五”科技重大专项（2008ZX10002）
