CAJ | 학술논문

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-IFN α-2a(180 μg/周)联合利巴韦林口服(15 μg·kg-1·d-1)抗病毒治疗.分别在0、4、12、24、48、72周时,检测患者血清中ALT、HCV RNA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one way ANOVA模块进行t检验;两组之间比较采用独立样本t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％±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,P值均＜0.05).正常人TLR3及PD1/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-IFN α-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、PD1/TLR4高表达,初治患者用Peg-IFN α-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.
목적 관찰취을이순간우소(Peg-IFN) α-2a연합리파위림치료만성병형간염초치환자항병독치료48주급정약후수방지72주외주혈단개핵세포(PBMC)표면정서성사망수체(PD)-1、Toll양수체(TLR)3、TLR4표체솔적변화,탐토PBMC표면PD-1、TLR3、TLR4시부삼여료숙주항HCV고유면역화획득성면역과정,급기여지속병독학응답적관계.방법 수집만성병형간염초치환자23례급정상인10명,만성병형간염환자균접수48주Peg-IFN α-2a(180 μg/주)연합리파위림구복(15 μg·kg-1·d-1)항병독치료.분별재0、4、12、24、48、72주시,검측환자혈청중ALT、HCV RNA변화;분리환자PBMC,류식세포술검측불동시간점환자화정상인PBMC표면PD-1、TLR3、TLR4적표체솔,분석지속병독학응답(SVR)여비SVR환자상술지표적변화화차이.다조자료지간적비교채용one way ANOVA모괴진행t검험;량조지간비교채용독립양본t검험.결과 병형간염환자기선시여정상인PBMC표면PD-1、TLR4、PDl/TLR4표체솔분별위:45.44％±7.32％、57.74％±15.45％、35.42％±7.97％여16.82％±4.13％、21.09％±2.89％、14.12％±2.89％,병형간염환자기선수평교정상인명현승고(t치분별위4.675,5.148,2.549,P치균＜0.05).정상인TLR3급PD1/TLR3표체솔교병형간염환자기선수평초저,단차이무통계학의의(P＞0.05).재Peg-IFN α-2a연합리파위림치료기간,만성병형간염환자PBMC상PD-1급TLR4표체솔축점하강,TLR3표체솔축점승고;차SVR환자PBMC상PD-1화TLR3표체솔적변화여비SVR환자적차이명현(P＜0.05).결론 만성병형간염환자PBMC상PD-1、TLR4、PD1/TLR4고표체,초치환자용Peg-IFN α-2a치료가이사저사지표표체하강,TLR3적표체솔승고;PBMC표면PD-1화TLR3적표체가능여환자출현SVR밀절상관.
Objective To investigate the dynamic changes in expression of programmed death (PD)-1,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-IFNα-2a) plus ribavirin (RBV) combination therapy,and to analyze 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-IFNα-2a (180 μg,subcutaneous injection,once weekly) plus RBV (15 μg/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 ofPD-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 oneway ANOVA.Results The expression ratios ofPD-1,TLR4 and PD-1:TLR4 on PBMCs were significantly higher in CHC patients before therapy than in the healthy controls (45.20 ± 7.12％ vs.16.82 ± 4.13％,58.45 ± 15.13％ vs.21.09 ± 2.89％,and 35.54 ± 7.69％ vs.14.12 ± 2.89％；all P ＜ 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-IFNα-2a plus RBV combination therapy,the expression ratios ofPD-1 and TLR4 on 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 ofPD-1,TLR4,and PD-1:TLR4 is up-regulated in the total PBMCs of CHC patients.PegIFNα-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 rehted to achievement of SVR.