聚乙二醇干扰素α-2a治疗慢性乙型肝炎过程中外周血γδT细胞的变化及其临床意义

Changes and clinical significance of γδT cells in peripheral blood of patients with chronic hepatitis B during pegylated interferon α-2a treatment

目的 观察慢性乙型肝炎（CHB）患者在聚乙二醇干扰素（Peg-IFN） α-2a治疗过程中外周血γδT细胞的变化,并分析其与临床指标及疗效的相关性. 方法 收集HBeAg阳性的CHB患者在Peg-IFNα-2a治疗不同时间点的外周全血,包括0周17例、12周20例、24周20例及48周16例,其中有11例患者在每个时间点均收集血样观察.流式细胞术检测外周血中γ δ T及其亚群细胞Vδ1T、Vδ2T、效应记忆γδT（γ δTem）、中央记忆γ δT （γ δTcm）、初始γδT （γδTnaive）与终末分化效应γ δT （γ δTeff）细胞的频率.同时检测肝功能、HBV血清病毒等标志物及HBV DNA水平.用SPSS 23.0统计软件分析比较各治疗时间点细胞比例的差异,以及细胞比例与丙氨酸氨基转移酶（ALT）、HBsAg、HBeAg或HBV DNA水平的相关性;并且分析持续随访的11例患者中,γδT及其亚群细胞比例与Peg-IFN α-2a治疗应答的相关性.结果 在Peg-IFNα-2a治疗0～ 48周期间,CHB患者外周血γδT及Vδ 2T细胞比例逐渐下降,48周时的γδT细胞和Vδ 2T细胞比例分别为6.89％（5％,8.15％）、4.61％ （2.16％,6.50％）,明显低于0周水平[12.5％（7.73％,19.00％）、6.59％ （3.86％,13.62％）],差异均具有统计学意义（P值均＜0.05）;但V δ 1T、γ δ Tem、γ δTcm、γ δTnaive或γ δTeff亚群细胞的比例在各时间点的差异均无统计学意义（P值均＞ 0.05）.同时ALT、HBsAg、HBeAg或HBV DNA水平与γδT或Vδ2T细胞比例均呈显著性正相关（P值均＜0.05）.11例持续随访患者中,应答者的γ δ Tem细胞的比例在各时间点均显著低于无应答者,差异均有统计学意义（P值均＜ 0.05）,而其他γδT亚群细胞比例在两组间的差异均无统计学意义（P值均＞ 0.05）. 结论 Peg-IFN α-2a治疗CHB过程中γδT细胞比例随着HBV复制降低及肝脏炎症缓解而呈下降趋势,且具有较低效应记忆γδT细胞比例的CHB患者在Peg-IFNα-2a治疗中可能更易获得应答.

Objective To observe the changes of γδT cells in the peripheral blood of patients with chronic hepatitis B （CHB） during pegylated interferon α-2a treatment,and to analyze the correlation between clinical indicators and curative effects.Methods Peripheral blood of hepatitis B e antigen （HBeAg）-positive CHB patients were collected at different time points of Peg-IFNα-2a treatment,including 17 patients at 0 weeks,20 patients at 12 weeks,20 patients at 24 weeks,and 16 patients at 48 weeks.From these 11 patients,blood samples were frequently observed at 0,12,24,and 48 weeks of treatment.The frequencies of γδT and its subpopulation cells Vδ1T,Vδ2T,effector memory γδT （γδTem）,central memory γδT （γδTcm）,initial γδT （γδTnaive） and terminal differentiation effect γδT （γδTeff） cells in peripheral blood were detected by flow cytometry.Liver function,serum HBV markers and HBV DNA levels were measured simultaneously.SPSS 23.0 statistical software was used to analyze the differences in cell proportions at each treatment time point,and the correlation between cell proportions and alanine aminotransferase （ALT）,HBsAg,HBeAg or HBV DNA levels.In addition,the correlation between the proportions ofγδT and its subpopulation cells and the response to Peg-IFNα-2a treatment in the 11 patients with continuous follow-up were analyzed.Results The percentage of γδT and Vδ2T cells in peripheral blood of patients with CHB decreased gradually during the period of 0-48 weeks of Peg-IFNα-2a treatment.The percentages of γδT cells and Vδ2T cells at 48 weeks were 6.89% （5%,8.15%）,4.61% （2.16%,6.50%）,respectively;significantly lower than the 0 week [12.5% （7.73%,19%）,6.59% （3.86%,13.62%）],the differences were statistically significant （P 〈 0.05）.The proportions of Vδ1T,γδTem,γδTcm,γδTnaive,or γδTeff subpopulations were not statistically different at each time points （all P 〉 0.05）.At the same time,the levels ofALT,HBsAg,HBeAg or HBV DNA were positively correlated with the ratio ofγδT or Vδ2T cells （P 〈 0.05）.Among the 11 patients with continuous followed-up,the proportion of γδTem cells in responders was significantly lower than that of non-responders at each time points,and the difference was statistically significant （P 〈 0.05）.There was no statistically significant difference between the two groups （all P 〉 0.05）.Conclusion The proportion ofγδT cells in the course of CHB treatment with Peg-IFNα-2a reduces the liver inflammation by decreasing the replication of HBV virus.Chronic hepatitis B patients with a lower proportion of effector memory （γδTem） cells may be more likely to get better response with Peg-IFNα-2a.