慢性乙型肝炎患者外周血CD4+CD25+CD127low/-调节性T细胞表达及其与肝组织病理、HBV-cccDNA的关系
Association of expression of peripheral CD4+CD25+CD127low/- regulatory T cells with liver pathology and HBV-cccDNA in patients with chronic hepatitis B

摘要：目的 探讨慢性乙型肝炎(CHB)患者外周血CD4+CD25+CD127low/-调节性T细胞与肝组织HBV-cccDNA及HBV-DNA载量、血清HBV-DNA载量、肝脏病理及肝功能的相关性。方法 采用流式细胞仪检测30例CHB及30例健康体检者外周血CD4+CD25+CD127low/-调节性T细胞占CD4+T细胞百分率；并对其中22例患者行肝组织活检，采用实时荧光定量聚合酶链反应(FQ-PCR)检测CHB患者肝组织HBV-cccDNA、HBV-DNA载量；肝组织进行苏木素-伊红(HE)染色、Masson染色、网染、VG染色，行炎症(G)分级及纤维化(S)分期；同期检测肝功能ALT水平，并对各检测值进行比较和相关性分析。结果 CHB患者外周血CD4+CD25+CD127low/-Treg细胞的百分率为(9.27±2.07)%，明显高于健康对照组(4.96±0.97)%，差异具有统计学意义(P<0.05)；CHB患者外周血CD4+CD25+CD127low/-Treg细胞与血清HBV DNA(lgIU/mL)载量呈正相关(Pearson相关系数r=0.38, P<0.05)；与肝组织HBV-cccDNA及HBV-DNA(lgcps/mg)均无明显相关性(Pearson相关系数r分别为0.162、0.306，两者均P>0.05)；与肝组织炎症级别及纤维化分期均无明显相关性(Spearman秩相关系数r分别为0.056、－0.047，两者均为P>0.05)；与ALT亦无明显相关性(Pearson相关系数r=0.044，P>0.05)。结论 慢性乙型肝炎患者外周血CD4+CD25+CD127low/-Treg细胞水平明显升高且与血清HBV-DNA载量呈正相关，提示Treg细胞在慢性乙型肝炎中发挥着重要的负性免疫调节作用，可能是导致病毒复制持续存在的重要因素。
ABSTRACT: Objective To investigate the expression level of CD4+CD25+CD127low/- regulatory T cells in the peripheral blood of patients with chronic hepatitis B (CHB) and its association with liver HBV-cccDNA and HBV-DNA, HBV-DNA of the peripheral blood, liver pathology and liver function. Methods We enrolled 30 CHB patients and 30 healthy controls. The percentage of peripheral blood CD4+CD25+CD127low/- regulatory T cells in CD4+ T cells were detected by flow cytometry. And 22 patients underwent liver biopsy; the quantities of intrahepatic HBV cccDNA and HBV DNA were detected by real-time fluorescent quantitative PCR. The liver tissue was dyed with HE dyeing, Masson dyeing, Nets dyeing and VG dyeing. Inflammatory grading and fibrosis staging were performed. The level of alanine aminotransferase (ALT) was collected simultaneously. Results The frequency of the peripheral blood CD4+CD25+CD127low/-Treg in CHB was (9.27±2.07)%, which was significantly higher than that in healthy controls [(4.96±0.97)%, P<0.05]. The peripheral blood CD4+CD25+CD127low/-？？ Treg in CHB was positively correlated with the plasma HBV DNA (lgIU/mL) (r=0.38, P<0.05), and there was no significant correlation with the level of liver inflammation or fibrosis stage (r=0.056, -0.047 respectively, both P>0.05). Also there were no significant correlation with ALT (r=0.044, P>0.05). Conclusion The expression of CD4+CD25+CD127low/- regulatory T cells in the peripheral blood of CHB patients increased significantly, which suggested that Tregs plays a negative role in immune regulation of CHB and may be an important factor of the persistence of viral replication