从Th1/Th2、Treg/Th17之间的平衡研究轻、中度慢性乙型肝炎患者CD4^+T淋巴细胞的免疫失衡状态

Balance among Th1,Th2,Th17 and Treg in mild to moderate chronic hepatitis B patients with immune imbalance of CD4^+ T cells

目的从Th17及其相关细胞因子与Th1、Th2、Treg的相互关系,探讨轻、中度CHB患者CD4^+T淋巴细胞的整体免疫失衡状态。方法选择轻、中度CHB患者,与健康志愿者进行比较,观察其外周血各免疫细胞(Th17、Th1、Th2、Treg、Th1/Th2、Treg/Th17)、相关细胞因子(IL-17、IL-6、TGF-β、IFN-γ、IL-4)、主要效应分子(穿孔素、颗粒酶B)和核转录因子(孤独核受体RORγt、Foxp3等)的变化及其相互关系。结果与健康志愿者相比,CHB患者CD8^+T淋巴细胞比例更高,CD4^+/CD8^+比例显著下降,差异有统计学意义(P<0.01);与健康志愿者相比,CHB患者Th1细胞比例更高,Th1/Th2显著升高,差异有统计学意义(P<0.01);与健康志愿者相比,CHB患者Th17细胞比例更高,Treg/Th17比例显著下降,差异有统计学意义(P<0.01);与健康志愿者相比,CHB患者穿孔素和颗粒酶B的表达更高,差异有统计学意义(P<0.01);与健康志愿者相比,CHB患者FOXP3、IL17mRNA和RORγt的表达差异无统计学意义(P>0.05);与健康志愿者相比,CHB患者TGF-B1、IL-6、IL-17显著增高,差异有统计学意义(P<0.01);IFN-γ和IL-4的表达差异无统计学意义(P>0.05)。结论轻、中度CHB患者在免疫细胞及其相关细胞因子、效应分子等各环节,尤其是Treg/Th17比例,均处于严重的免疫失衡状态。

Objective To investigate the immune imbalance of CD4＋ T cells in patients with mild to moderate chronic hepatitis B （CHB） through exploring the relationship of the T helper cell 17 （Th17） and the related cytokines with T helper cell 1 （Th1） , T helper cell 2 （Th2） and regulatory T Cell （Treg）. Methods Seventy mild to moderate CHB patients and 30 healthy volunteers were enrolled, respectively. Immune cells （Th17, Th1, Th2, Treg, Th1/Th2, Treg/Th17） and main effector molecules （perforin and granzyme B） in peripheral blood were measured using flow cytometry. The related cytokines including interleukin （IL）-17, IL-6, transforming growth factor （TGF） （3, interferon （IFN） γ and IL-4 were detected using enzyme linked immunosorbent assay （ELISA）. Nuclear transcription factor including lonely nuclear receptor （RORγt） , Foxp3 were measured using polymerase chain reaction （PCR）. Results Compared with those in healthy volunteers,percentage of CD8＋ T cells was higher but CD4＋ /CD8＋ was markedly decreased in CHB patients （P〈0. 01）. Levels of Th1, Th1/Th2 and Th17 were significantly increased in CHB patients, while levels of Th2, Treg and Treg/Th17 were decreased compared with control group （P〈0. 01）. Perforin and granzyme B expression were higher in CHB patients （P〈0. 01）. No significant difference was observed in the expression of Foxp3, IL17mRNA and RORγt between healthy volunteers and CHB patients （P〉0. 05）. Compared with healthy volunteers, expressions of TGF-B1, IL-6 and IL-17 were increased significantly in CHB patients （P〈0. 01） , while there was no significant difference in the expression of IFN-γ and IL-4 （ 〉0. 05）. Conclusion In mild to moderate CHB patients, the chain of immunity, including immune cells, related cytokines and effector molecules, was in a serious imbalance immune state, especially the ratio of Treg/Th17.