慢性乙型肝炎患者外周血中调节性T淋巴细胞、Ⅰ型调节性T淋巴细胞和辅助性T淋巴细胞17的表达特点及临床意义

摘要

目的：探讨慢性乙型肝炎（CHB）患者外周血中CD4＋CD25＋Foxp3＋（Treg）、CD4＋CD25＋IL－10＋（Tr1）和CD4＋IL－17＋（Th17）的表达特点及其与CHB病情的关系。方法选取2013年1月至2014年6月解放军第一八〇医院收治的轻中度CHB患者和重度CHB患者各40例，另选取同期无肝病诊断史的健康人40例为对照。分别抽取CHB患者及对照者的外周血，以流式细胞术检测外周血中 Treg、Tr1和 Th17表达水平，real time －PCR 检测血清中 Foxp3、RORγt mRNA 水平，ELISA 检测患者血清中TGFβ1、IL－10、IL－17表达水平。计量资料组间比较采用单因素方差分析和LSD－t检验。结果轻中度和重度CHB患者与对照组相比，Treg、Tr1和Th17的百分率，转录因子Foxp3、RORγt表达水平，细胞因子TGFβ1、IL－10、IL－17水平均明显高于健康对照组（P值均＜0．05）；而Treg／Th17和Foxp3／RORγt值显著低于对照组（P值均＜0.05）。与轻中度CHB患者相比，重度CHB患者Treg、Tr1和Th17的百分率，Foxp3、RORγt表达水平，TGFβ1、IL－10、IL－17水平均显著升高（P值均＜0．05）；Treg／Th17和Foxp3／RORγt值显著降低（P值均＜0．05）。结论 CHB患者中Treg（特别是Tr1）和Th17的表达失衡程度与CHB病情进展呈正相关，因此，对于CHB患者，纠正Treg（特别是Tr1）和Th17表达失衡可能是很重要的治疗措施，同时监测Treg（特别是Tr1）和Th17的表达比例改变，对于估计CHB患者病情预后、指导临床治疗有积极作用。%Objective To investigate the expression characteristics of Treg,Tr1,and Th17 cells in the peripheral blood of patients with chronic hepatitis B (CHB)and their correlations with the state of CHB.Methods A total of 40 patients with mild-to-moderate CHB and 40 patients with severe CHB who were admitted to The 180th Hospital of PLA from January 2013 to June 2014 were included into this study, and 40 healthy people with normal liver function were recruited as control group.Peripheral blood was collected from CHB patients and healthy controls,and then the percentages of Treg,Tr1,and Th17 cells in peripheral blood were determined by flow cytometry.The levels of Foxp3 mRNA and RORγt mRNA in serum were measured by real-time PCR,and the expression levels of TGF-β1,IL-10,and IL-17 were measured by ELISA.Comparison of continuous data between the groups was made by one-way ANOVA analysis and LSD-t test. Results (1)Compared with the control group,the two groups of CHB patients had significantly higher percentages of Tr1,Treg,and Th17 cells (P<0.05),and had significantly higher levels of Foxp3 mRNA and RORγt mRNA (P<0.05)and levels of TGF-β1,IL-10,and IL-17 (P<0.05),while the two groups of CHB patients had significantly lower Treg/Th17 ratio (P<0.05)and Foxp3/RORγt ratio (P<0.05)than the healthy controls.(2)The patients with severe CHB had significantly higher percentages of Treg,Tr1,and Th17 cells (P<0.05),and had significantly higher levels of Foxp3 mRNA and RORγt mRNA (P<0.05)and levels of TGF-β1,IL-10,and IL-17 (P<0.05),as compared with the patients with mild-to-moderate CHB,while the patients with severe CHB had significantly lower Treg/Th17 ratio (P<0.05)and Foxp3/RORγt ratio (P<0.05)than those with mild-to-moderate CHB.Conclusion The expression imbalance of Treg cells (especially Tr1 cells)and Th17 cells in CHB patients is positively correlated with the progression of the disease;therefore,it may be important to correct the expression imbalance of Treg cells and Th17 cells in the treatment of CHB.Meanwhile,monitoring the percentages of Treg cells (especially Tr1 cells)and Th17 cells is helpful for assessing the clinical outcomes of CHB patients and guiding the clinical treatment.