慢病毒载体介导Runx3基因过表达对慢性乙型肝炎患者Th细胞分化的影响

Effects of lentiviral vector-mediated Runx3 overexpression on Th cell differentiation in patients with chronic hepatitis B

目的探讨Runx3基因过表达对慢性乙型肝炎(chronic hepatitis B,CHB)患者Th细胞分化的影响。方法重组慢病毒载体p GC-FU-Runx3与阴性对照慢病毒载体p GC-FU分别转染29例CHB患者外周血CD4+T细胞,分离培养5 d的细胞培养液上清,应用ELISA检测Th1型细胞因子IFN-γ和Th2型细胞因子IL-4的表达水平;收集培养5 d的CD4+T细胞,应用实时定量PCR检测转录因子T-bet、GATA3m RNA的表达水平。结果 1与p GC-FU转染组比较,p GC-FU-Runx3转染组Th1型细胞因子IFN-γ的表达水平明显升高(P=0.003)。2与p GC-FU转染组比较,p GC-FU-Runx3转染组Th2型细胞因子IL-4的表达水平明显降低(P=0.007)。3与p GC-FU转染组比较,p GC-FU-Runx3转染组IFN-γ/IL-4比值明显增大(P=0.001)。4与p GC-FU转染组比较,p GC-FU-Runx3转染组T-bet、GATA3 m RNA的表达水平无显著性差异(P均>0.05),但p GC-FU-Runx3转染组T-bet/GATA3比值较p GC-FU转染组明显增大(P=0.005)。结论 Runx3过表达可促进CHB患者Th1型细胞因子的分泌,抑制Th2型细胞因子分泌,促使CHB患者外周血Th细胞向Th1细胞分化,使其Th1/Th2失平衡得到改善。

Objective To investigate the effect of lentiviral vector-mediated Runx3 overexpression on Th cell differentiation in patients with chronic hepatitis B （CHB）. Methods Peripheral CD4＋ T cells derived from 29 CHB patients were transfected with the recombinant lentiviral vector （pGC-FU-Runx3） and the negative control lentiviral vector （pGC-FU）, respectively. Then the cell culture supematants and the CD4＋ T cells were collected on the 5th day. The expression of Thl-type cytokine （IFN-y） and Th2-type cytokine （IL-4） were measured by ELISA and the expression of T-bet and GATA3 mRNA were assayed by quantitative real- time PCR. Results ①Compared with the pGC-FU transfected group, the expression of IFN-y in the pGC- FU-Runx3 transfected group significantly increased （P = 0.003）. ②Compared with the pGC-FU transfected group, the expression of IL-4 in the pGC-FU-Runx3 transfected group significantly decreased （P = 0.007）. ③Compared with the pGC-FU transfected group, the ratio of IFN-y/IL-4 in the pGC-FU-Runx3 transfected group significantly increased （P = 0.001）. ④There was no difference in T-bet and GATA3 mRNA expression between the pGC-FU transfected group and the pGC-FU-Runx3 transfected group （both P 〉 0.05）. However, compared with the pGC-FU transfected group, the ratio of T-bet/GATA3 in the pGC-FU-Runx3 transfected group significantly increased （P = 0.005）. Conclusions Runx3 overexpression can promote the secretion of Th 1-type cytokines and inhibit the secretion of Th2-type cytokines in CHB patients. It can also induce Th cell differentiating into Thl cell lineage and improve the Thl/Th2 imbalance in CHB patients.