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目的:观察急性HBV感染患者外周血FoxP3+CD4+CD25+调节性T细胞(Treg)的数量和功能的变化特点。方法:采集15例急性乙型肝炎(AHB)患者急性期、恢复早期及痊愈后外周血、15例健康人的外周血,分离PBMC,通过MACS免疫磁珠分选CD4+T细胞和Treg,应用实时荧光定量RT-PCR比较AHB患者不同疾病发展阶段(急性期,恢复早期,痊愈后)CD4+T细胞的FoxP3(Forkhead/winged helix tran-scription factor)mRNA表达量的差异,应用3[H]掺入法检测Treg抑制CD4+CD25-T细胞增殖的能力的差异。所有病例及对照均ELISA检测HBsAg、HBsAb、HBeAg、HBeAb、HB-cAb,实时荧光定量RT-PCR检测血清HBV DNA载量,同时进行肝功能检测。结果:AHB患者CD4+T细胞表达FoxP3mRNA的量在急性期稍低于健康对照,在疾病恢复早期明显高于健康对照(t=3.44,P<0.01),痊愈后恢复至正常。Treg抑制CD4+CD25-T细胞增殖的能力在急性期显著低于恢复早期(t=-5.30,P<0.01)和痊愈后(t=-3.20,P<0.05)。结论:Treg在AHB患者急性期数量减少,功能减弱,在恢复早期数量增加,功能增强,提示Treg可能在急性肝炎的不同发病阶段中通过调节细胞免疫反应影响乙肝病毒清除。
Objective: To observe the changes of the number and function of Foxp3 + CD4 + CD25 + regulatory T cells (Tregs) in peripheral blood of patients with acute HBV infection. Methods: Peripheral blood of 15 patients with acute hepatitis B (AHB) were collected from acute phase of acute hepatitis B (AHB). Peripheral blood of 15 healthy volunteers were recovered from early and recovered peripheral blood. PBMCs were isolated. CD4 + T cells and Tregs were sorted by MACS magnetic beads. Real-time fluorescent quantitative RT-PCR was used to compare the mRNA expression levels of FoxP3 (Foxp3 / Foxp3) in AHB patients with different stages of disease (acute stage, early recovery and recovery) Differences in the ability of Tregs to inhibit the proliferation of CD4 + CD25-T cells were examined by incorporation. All cases and controls were detected by ELISA HBsAg, HBsAb, HBeAg, HBeAb, HB-cAb, real-time fluorescence quantitative RT-PCR detection of serum HBV DNA load, and liver function tests. Results: The amount of FoxP3 mRNA expression in CD4 + T cells in AHB patients was slightly lower than that in healthy controls at acute stage, significantly higher than that in healthy controls (t = 3.44, P <0.01) during the early stage of AHB recovery and returned to normal after recovery. The ability of Treg to inhibit the proliferation of CD4 + CD25-T cells was significantly lower in the acute phase than in the early recovery phase (t = -5.30, P <0.01) and after the recovery (t = -3.20, P <0.05). CONCLUSION: The number of Tregs is decreased, the function is weakened, the number of Tregs is increased in the early stage of recovery, and the function of Tregs is increased in the acute phase of AHB. It suggests that Treg may affect the clearance of HBV by regulating the cellular immune response in different stages of acute hepatitis.