nTreg与基因1型CHC患者抗病毒疗效的关系

Natural regulatory T cells in patients with chronic hepatitis C genotype 1 treated by pegylated interferon and ribavirin

目的研究基因1型慢性丙型肝炎(CHC)患者治疗过程中外周血自然调节性T细胞(nTreg)的动态变化及其与抗病毒疗效的关系。方法 32例初治的CHC患者,均为基因1型,治疗方案为干扰素联合利巴韦林治疗48周。分别在治疗前、治疗过程中2、4、8、12、24、48周以及治疗结束后12或24周采集外周血标本。用流式的方法检测外周血单个核细胞(PBMC)中nTreg的比率。COBASAmpliprep/COBASTaqmanHCV试剂盒检测HCV-RNA定量。结果 24例患者治疗12周时HCV-RNA转阴,并且随访至治疗结束后24周仍阴性,达到持续病毒学应答(SVR);8例患者治疗12周时病毒量未转阴,无应答(NVR),24周时终止治疗。应答组和无应答组治疗前外周血中nTreg的比率差异无统计学意义(P>0.05)。应答组治疗过程中2、4、12、24、48周及治疗结束后各个时间点PBMC中nTreg水平分别为(0.91±0.22)%、(1.31±0.29)%、(1.78±0.43)%、(1.92±0.44)%、(1.90±0.37)%、(1.14±0.35)%;在治疗2周(P<0.001)及治疗结束后(P<0.001)下降明显。无应答组治疗过程中2、4、12、24周及治疗结束后各时间点PBMC中nTreg水平分别为(1.21±0.15)%、(1.24±0.18)%、(1.42±0.11)%、(1.45±0.11)%、(1.14±0.10)%;Treg水平在治疗4周时缓慢上升(P=0.005),在治疗结束后显著下降,与治疗24周比较差异有统计学意义(P<0.001)。结论 CHC患者外周血nTreg水平与干扰素/利巴韦林抗病毒治疗的疗效密切相关,抗病毒治疗2周时nTreg水平的下降可以预测其疗效。提示nTreg可能在机体清除病毒的早期过程中起着重要作用。

Objective To explore the alteration of circulating natural regulatory T ceils （nTregs） in patients with chronic hepatitis C genotype 1 during pegylated interferon/ribavirin combination therapy and its clinical significance. Methods 32 chronic hepatitis C naive patients who had high viral titers of HCV genotype 1 and were treated with pegylated interferon plus ribavirin. Blood samples were collected at pre-treatment, week 2, week 4,week 8, week 12, week 24 and week 48 during therapy,and at week 12 or 24 of follow-up after termination of therapy. The frequency of CD4（＋）CD25（＋）Foxp3 （＋）Tregs in peripheral blood mononuclear ceils （PBMCs） was determined by flow cytometry. HCV-RNA was detected by Roche COBAS Ampliprep/COBAS Taqman HCV test. Results 24 patients with undetectable HCV-RNA at treatment week 12 achieved sustained virologic response （SVR）.8 patients with detectable HCV RNA at week 12 failed to achieve virologic response at week 24 and discontinue the treatment at week 24. There was no significant difference in the frequency of CD4（＋）CD25 （＋）Foxp3 （＋）Tregs in PBMCs（P〉0.05） at pre-treatment between the response group and nonresponse group. The frequencies of CD4 （＋）CD25 （＋）Foxp3 （＋） Tregs in PBMCs from the response group at week 2, week 4, week 12, week 24 and week 48 during therapy, and at follow-up were （0.91±0.22）%, （1.31±0.29）%, （1.78± 0.43）%, （ 1.92±0.44）%, （ 1.90±0.37）%, （ 1.14±0.35）%, respectively, obviously decreased at week 2（P〈0.001 ） and at follow-up（P〉0.05） and increased at week 4 （P〈0.001） and 12 （P〈0.001） as regard to previous adjacent time points. The frequencies of CD4 （＋）CD25 （＋）Foxp3 （＋） Tregs in PBMCs from the nonresponse group at week 2, week 4, week 12, week 24 during therapy, and at follow-up were （ 1.21±0.15）%, （ 1.24±0.18）%, （ 1.42±0.11 ）%, （1.45±0.11）%, （1.14±0.10） %, respectively, slightly increased （P=0.005） at week 4 during therapy and obviously decreased at follow- up as regard to previous adjacent time points （P〈0.001）. Conclusions The kinetics of nTregs during combination therapy of the response group was different from that of the nonresponse group. Significant decline of nTregs at treatment week 2 has close relationship with the early response to ribavirin/pegylated interferon combination therapy. This indicated that nTregs may play a very important role in the early clear of the virus.