干扰素治疗慢性乙肝患者外周血调节性T细胞动态变化和临床意义

目的探讨干扰素抗病毒治疗过程中慢性乙肝患者外周血CD4＋CD25＋调节性T细胞（CD4＋CD25＋Treg）的变化及其临床意义。方法采集干扰素抗病毒治疗的CHB患者46例外周血,在治疗前及治疗后12、24、36、48周,停药后24周时,分别以流式细胞仪检测外周血CD4＋CD25＋Treg比例,实时荧光定量RT-PCR检测血清HBV-DNA载量,酶联免疫吸附法检测HBV标志物,全自动生化分析仪检测ALT水平。结果在46例CHB患者中,完全应答的13例患者CD4＋CD25＋Treg频率在12、24、36、48周及停药后24周时分别为（4.16±0.64）%、（3.98±0.75）%、（3.70±0.55）%、（3.44±0.48）%、（3.07±0.48）%,呈逐步下降趋势,明显低于治疗前（5.73±0.62）%（P〈0.01）。部分应答的18例患者CD4＋CD25＋Treg频率在12、24、36、48周及停药后24周时分别为（4.98±0.62）%（、4.50±0.54）%、（4.10±0.64）%、（4.02±0.64）%（、3.82±0.47）%,呈逐步下降趋势,明显低于治疗前（5.84±0.68）%（P〈0.01）。无应答的15例患者CD4＋CD25＋Treg频率较治疗前无明显变化。完全应答、部分应答组患者Treg频率在12、24、36、48周及停药后24周时与无应答组比较,差异有统计学意义（P〈0.01）。治疗24、48周,停药后24周时,HBeAg阴转率分别为10.8%、26.1%、28.3%,发生HBeAg血清学转换者在治疗12周时CD4＋CD25＋Treg比例明显下降。结论本实验初步认为,干扰素抗病毒治疗过程中及治疗后,有应答的CHB患者的外周血CD4＋CD25＋Treg频率下降。治疗早期CD4＋CD25＋Treg比例下降的患者HBeAg血清学转换率可能更高。

Dynamic changes and clinical significance of regulatory T cells in peripheral blood after interferon therapy in chronic hepatitis B patients

Objective To explore the changes and clinical significance of CD4＋CD25＋regulatory T cells（CD4＋CD25＋Treg） in peripheral blood of chronic hepatitis B（CHB） patients during interferon therapy.Methods In this study,46 chronic hepatitis B patients with positive HBeAg were treated with interferon.Before and 12,24,36,48 weeks after treatment,and 24 weeks after drug withdrawal,flow cytometry was used to detect the proportion of peripheral blood CD4＋CD25＋Treg.Real-time RT-PCR was used to detect the levels of HBV-DNA in serum.Markers of HBV were detected by ELISA and the levels of alanine aminotransferase（ALT） were detected by full automatic biochemical analyzer.Results The frequencies of CD4＋CD25＋Treg reduced gradually in 13 complete responders of 46 CHB patients at week 12,24,36 and 48,and 24 weeks after drug withdrawal,were（4.16±0.64）%,（3.98±0.75）%,（3.70±0.55）%,（3.44±0.48）%,and（3.07±0.48）% respectively,which were significantly lower than baseline level（5.73±0.62）%（P0.01）.The decreasing frequencies of CD4＋CD25＋Treg in 18 partial responders were（4.98±0.62）%,（4.50±0.54）%,（4.10±0.64）%,（4.02±0.64）%,and（3.82±0.47）% respectively,which were significantly lower than baseline value（5.84±0.68）%（P0.01）.No significant changes in CD4＋CD25＋Treg were found in non-respondent group before and after treatment（P0.05）.Compared with the non-respondent group,the Treg frequencies of the fully respondent group and partially respondent group showed statistically significant differences at week 12,24,36 and 48,and 24 weeks after drug withdrawal（P0.01）.HBeAg seroconversion rates were 5（10.8%）,12（26.1%） and 13（28.3%） respectively at week 24 and 48 of treatment and 24 weeks after drug withdrawal.The frequency of CD4＋CD25＋Treg was reduced significantly in patients with HBeAg seroconversion at week 12.Conclusion Interferon therapy could reduce the frequency of CD4＋CD25＋Treg in peripheral blood of CHB patients responding to treatment.There is a higher HBeAg seroconversion rate in the patients with reduction in proportion of CD4＋CD25＋Treg at the early stage of treatment.