CAJ | 학술논문

目的观察慢性丙型肝炎(CHC)患者抗病毒治疗24周时外周血CD4+和CD8+T淋巴细胞(T细胞)表面表面程序性死亡1 (PD-1)和程序性死亡配体1(PD-L1)表达水平,分析其与抗病毒治疗临床转归的关系.方法 24例CHC患者,均采用聚乙二醇干扰素α-2a (Peg-IFN α-2a)每周皮下注射一次,联合利巴韦林800 ～ 1200 mg/d,治疗24 ～ 48周.采用流式细胞术和实时荧光定量检测患者治疗前、治疗4、12、24周外周血CD4+和CD8+T细胞表面PD-1、PD-L1表达水平和外周血HCV RNA,全自动生化分析仪检测ALT.采用SPSS16.0软件.两样本计量结果分析采用t检验,治疗前后的计量结果采用重复测量的单因素或两因素方差分析,所有检验为双侧检验. 结果 CHC患者治疗后4周HCV RNA阴性者19例,CD4+和CD8+T细胞表面PD-1的表达率在治疗前分别为18.6％±6.1％和16.6％±13.8％,治疗24周时分别为10.3％±7.7％和9.4％±4.6％,治疗前后比较,PD-1的表达明显下降,F值为12.406和4.955,P值为0.002和0.039,差异有统计学意义.CD8+T细胞表面PD-L1的表达率在治疗前为17.5％±13.7％,治疗4、12、24周时分别为25.9％±11.1％、29.6％±15.1％、32.0％±15.7％,治疗后明显升高,F值分别为9.063、8.365、9.736,P值均＜0.01.治疗4周时,HCV RNA阳性者5例,仅发现CD8+T细胞表面PD-L1的表达治疗24周(39.2％±15.6％)与治疗前(17.4％±16.7％)比较明显升高,F=10.292,P=0.033.持续病毒学应答者20例:CD4+T细胞表面PD-1的表达在治疗4、12、24周分别为14.4％±7.5％、14.0％±6.9％、10.7％±7.6％,治疗前为20.2％±7.5％,与治疗前比较明显下降,F值分别为6.133、5.541、14.780,P＜0.05或P＜0.01.CD8+T细胞表面PD-1的表达在治疗12、24周分别为10.2％±4.6％和10.1％±4.9％,治疗前为16.8％±13.4％,治疗前后比较,PD-1的表达在治疗后明显下降,F值为4.964和4.613,P值均＜0.05.CD8+T细胞表面PD-L1的表达在治疗12、24周分别为30.8％±16.6％和35.2％±16.5％,治疗前为19.0％±14.5％,治疗后明显升高,F=6.442,P=0.020和F=12.349,P=0.002.复发组4例,各治疗时间点PD-1和PD-L1与治疗前比较,差异无统计学意义.结论快速有效的抗病毒治疗可以下调CHC患者外周血CD4+和CD8+T细胞表面PD-1的表达,上调CD8+T细胞表面PD-L1的表达.CHC患者外周血CD4+和CD8+T细胞表面PD-1和PD-L1表达水平的变化可能与患者抗病毒治疗临床转归存在关系. 目的觀察慢性丙型肝炎(CHC)患者抗病毒治療24週時外週血CD4+和CD8+T淋巴細胞(T細胞)錶麵錶麵程序性死亡1 (PD-1)和程序性死亡配體1(PD-L1)錶達水平,分析其與抗病毒治療臨床轉歸的關繫.方法 24例CHC患者,均採用聚乙二醇榦擾素α-2a (Peg-IFN α-2a)每週皮下註射一次,聯閤利巴韋林800 ～ 1200 mg/d,治療24 ～ 48週.採用流式細胞術和實時熒光定量檢測患者治療前、治療4、12、24週外週血CD4+和CD8+T細胞錶麵PD-1、PD-L1錶達水平和外週血HCV RNA,全自動生化分析儀檢測ALT.採用SPSS16.0軟件.兩樣本計量結果分析採用t檢驗,治療前後的計量結果採用重複測量的單因素或兩因素方差分析,所有檢驗為雙側檢驗. 結果 CHC患者治療後4週HCV RNA陰性者19例,CD4+和CD8+T細胞錶麵PD-1的錶達率在治療前分彆為18.6％±6.1％和16.6％±13.8％,治療24週時分彆為10.3％±7.7％和9.4％±4.6％,治療前後比較,PD-1的錶達明顯下降,F值為12.406和4.955,P值為0.002和0.039,差異有統計學意義.CD8+T細胞錶麵PD-L1的錶達率在治療前為17.5％±13.7％,治療4、12、24週時分彆為25.9％±11.1％、29.6％±15.1％、32.0％±15.7％,治療後明顯升高,F值分彆為9.063、8.365、9.736,P值均＜0.01.治療4週時,HCV RNA暘性者5例,僅髮現CD8+T細胞錶麵PD-L1的錶達治療24週(39.2％±15.6％)與治療前(17.4％±16.7％)比較明顯升高,F=10.292,P=0.033.持續病毒學應答者20例:CD4+T細胞錶麵PD-1的錶達在治療4、12、24週分彆為14.4％±7.5％、14.0％±6.9％、10.7％±7.6％,治療前為20.2％±7.5％,與治療前比較明顯下降,F值分彆為6.133、5.541、14.780,P＜0.05或P＜0.01.CD8+T細胞錶麵PD-1的錶達在治療12、24週分彆為10.2％±4.6％和10.1％±4.9％,治療前為16.8％±13.4％,治療前後比較,PD-1的錶達在治療後明顯下降,F值為4.964和4.613,P值均＜0.05.CD8+T細胞錶麵PD-L1的錶達在治療12、24週分彆為30.8％±16.6％和35.2％±16.5％,治療前為19.0％±14.5％,治療後明顯升高,F=6.442,P=0.020和F=12.349,P=0.002.複髮組4例,各治療時間點PD-1和PD-L1與治療前比較,差異無統計學意義.結論快速有效的抗病毒治療可以下調CHC患者外週血CD4+和CD8+T細胞錶麵PD-1的錶達,上調CD8+T細胞錶麵PD-L1的錶達.CHC患者外週血CD4+和CD8+T細胞錶麵PD-1和PD-L1錶達水平的變化可能與患者抗病毒治療臨床轉歸存在關繫.
목적 관찰만성병형간염(CHC)환자항병독치료24주시외주혈CD4+화CD8+T림파세포(T세포)표면표면정서성사망1 (PD-1)화정서성사망배체1(PD-L1)표체수평,분석기여항병독치료림상전귀적관계.방법 24례CHC환자,균채용취을이순간우소α-2a (Peg-IFN α-2a)매주피하주사일차,연합리파위림800 ～ 1200 mg/d,치료24 ～ 48주.채용류식세포술화실시형광정량검측환자치료전、치료4、12、24주외주혈CD4+화CD8+T세포표면PD-1、PD-L1표체수평화외주혈HCV RNA,전자동생화분석의검측ALT.채용SPSS16.0연건.량양본계량결과분석채용t검험,치료전후적계량결과채용중복측량적단인소혹량인소방차분석,소유검험위쌍측검험. 결과 CHC환자치료후4주HCV RNA음성자19례,CD4+화CD8+T세포표면PD-1적표체솔재치료전분별위18.6％±6.1％화16.6％±13.8％,치료24주시분별위10.3％±7.7％화9.4％±4.6％,치료전후비교,PD-1적표체명현하강,F치위12.406화4.955,P치위0.002화0.039,차이유통계학의의.CD8+T세포표면PD-L1적표체솔재치료전위17.5％±13.7％,치료4、12、24주시분별위25.9％±11.1％、29.6％±15.1％、32.0％±15.7％,치료후명현승고,F치분별위9.063、8.365、9.736,P치균＜0.01.치료4주시,HCV RNA양성자5례,부발현CD8+T세포표면PD-L1적표체치료24주(39.2％±15.6％)여치료전(17.4％±16.7％)비교명현승고,F=10.292,P=0.033.지속병독학응답자20례:CD4+T세포표면PD-1적표체재치료4、12、24주분별위14.4％±7.5％、14.0％±6.9％、10.7％±7.6％,치료전위20.2％±7.5％,여치료전비교명현하강,F치분별위6.133、5.541、14.780,P＜0.05혹P＜0.01.CD8+T세포표면PD-1적표체재치료12、24주분별위10.2％±4.6％화10.1％±4.9％,치료전위16.8％±13.4％,치료전후비교,PD-1적표체재치료후명현하강,F치위4.964화4.613,P치균＜0.05.CD8+T세포표면PD-L1적표체재치료12、24주분별위30.8％±16.6％화35.2％±16.5％,치료전위19.0％±14.5％,치료후명현승고,F=6.442,P=0.020화F=12.349,P=0.002.복발조4례,각치료시간점PD-1화PD-L1여치료전비교,차이무통계학의의.결론 쾌속유효적항병독치료가이하조CHC환자외주혈CD4+화CD8+T세포표면PD-1적표체,상조CD8+T세포표면PD-L1적표체.CHC환자외주혈CD4+화CD8+T세포표면PD-1화PD-L1표체수평적변화가능여환자항병독치료림상전귀존재관계.
Objective To evaluate the changes in programmed death 1 (PD-1) and programmed death ligand 1 (PD-L1) expression on peripheral blood T lymphocytes of patients with chronic hepatitis C (CHC) over the 24 weeks course of antiviral therapy.Methods Twenty-four CHC patients administered 24 weeks of combination antiviral therapy with pegylated-interferon-alpha-2a (Peg-IFNα-2a) and ribavirin (RBV) were enrolled for study from the Nanjing Second Hospital between October 2008 and October 2011.Peripheral blood was collected before treatment initiation,at treatment weeks 4,12 and 24,and post-treatment week 24 (to investigate sustained virologic response (SVR),and used to measure expression of PD-1 and PD-L1 on CD4+ and CD8+ T lymphocytes by flow cytometry,load of serum hepatitis C virus (HCV) RNA by real-time polymerase chain reaction,and level of serum alanine aminotransferase (ALT) by auto-biochemical analyzer.Intergroup differences were analyzed by the two-sample t-test,and the significance of differences between preand post-treatment measurements was determined by one-way or two-way repeated measurements analysis of variance tests.Results At treatment week 4,19 of the CHC patients were HCV RNA-negative.Among those patients the PD-1 expression on both T lymphocyte subsets showed a significant decrease from pre-treatment to post-treatment week 24 (CD4+:18.6 ± 6.1％ vs.10.3 ± 7.7％,F=12.406,P =0.002; CD8+:16.6 ± 13.8％ vs.9.4 ± 4.6％,F =4.955,P =0.039).However,the CD8+ lymphocyte subset showed significant increase in PDL1 expression during treatment (pre-treatment:17.5 ± 13.7％ vs.treatment week 4:25.9 ± 11.1％,F =9.063,P ＜ 0.01; 12:29.6 ± 15.1％,F =8.365,P ＜ 0.01; 24:32.0 ± 15.7％,F =9.736,P ＜ 0.01).Among the five CHC patients showing HCV RNA-positivity at treatment week 4 there was only a significant difference observed in the increased expression of PD-L1 on CD8+ lymphocyte subset from pre-treatment to treatment week 24 (17.4 ± 16.7％ vs.39.2 ± 15.6％,F=10.292,P =0.033).Twenty of the CHC patients achieved SVR.among whom the PD-1 expression was significantly decreased during treatment on the CD4+ lymphocyte subset (pretreatment:20.2 ± 7.5％ vs.treatment week 4:14.4 ± 7.5％,F =6.133,P ＜ 0.05; 12:14.0 ± 6.9％,F =5.541,P ＜0.05; 24:10.7 ± 7.6％,F =14.780,P ＜ 0.05) and on the CD8+ lymphocyte subset (pre-treatment:16.8 ± 13.4％ vs.treatment week 12:10.2 ± 4.6％,F =4.964,P ＜ 0.05; 24:10.1 ± 4.9％,F =4.613,P ＜ 0.05).Additionally,the PD-L1 expression was significantly increased during treatment on the CD8+ lymphocyte subset (pre-treatment:19.0 ± 14.5％ vs.treatment week 12:30.8 ± 16.6％,F=6.442,P=0.020; 24:35.2 ± 16.5％,F=12.349,P=0.002).Among the four CHC patients who relapsed there were no significant differences observed in the expressions of PD-1 or PD-L1 on the CD4+ or CD8+ T lymphocytes.Conclusion The standard Peg-IFNα-2a + RBV combination antiviral therapy reduces PD-1 expression on CD4+ and CD8+ T lymphocytes and increases PD-L1 expression on CD8+ T lymphocytes in peripheral blood.The clinical outcome of CHC patients may be related to the antiviral therapy-induced changes in expressions of PD-1 and PD-L1 on T lymphocytes.