目的观察慢加急性乙型肝炎肝衰竭(Acute-on-chronic hepatitis B liver failure,ACHBLF)患者外周血和肝组织树突状细胞及其亚群的分布特征。方法选择11例进行肝移植的ACHBLF、10例慢性乙型肝炎患者和5例健康人,取外周血和肝组织,使用流式细胞仪计数外周血单个核细胞(PBMCs)和肝内浸润淋巴细胞(LILs)中浆样树突细胞(plasmacytoid DC,DCs)和髓样树突细胞(myeloid DC,mDCs)的频率,应用polyI:C和CpG2216刺激培养PBMCs和LILs,采用ELISA法检测上清IFN-α、IL-12和IL-10水平;采用免疫组化技术检测肝内pDC和mDC的分布。结果 ACHBLF患者肝内pDCs与外周血pDCs频率具有显著性差异(P〈0.05),而肝内mDC的频率与外周血之间差异无统计学意义(P〉0.05);ACLF患者经刺激的PBMCs和LILs能够分泌IFN-α和IL-12,但LILs分泌细胞因子的水平较PBMCs明显降低,而分泌IL-10却较PBMCs明显升高;ACHBLF患者肝组织浸润了大量的pDCs和mDCs。结论 ACHBLF患者肝内浸润大量的DC亚群细胞,并优先分泌IL-10,在发病机制中可能具有局部的免疫调节作用。
Objective To characterize the dendrictic cell subsets in peripheral blood and liver tissues of patients with hepatitis B-related acute-on-chronic liver failure(ACHBLF).Methods 11 patients with ACHBLF who had undergone liver transplantation,10 patients with chronic hepatitis B and 5 healthy persons were enrolled in this study,and their liver tissue samples and peripheral blood were collected.The proportion of myeloid DC(mDC)and plasmatoid DC(pDC)in peripheral blood mononuclear cells(PBMCs) and liver-infiltrating lymphocytes(LILs) were detected by flow cytometric analysis.The IFN-alpha,IL-12 and IL-10 levels in the supernatants by PBMCs and LILs were investigated by ELISA when they were stimulated with polyI:C or CpG2216,respectively.The prevalence of pDC and mDC in liver tissues were also assessed by immunohistochemical techniques.Results The number of intrahepatic pDCs was significantly higher than in peripheral blood in ACHBLF patients,but no difference was found between in mDC and PBMCs;The IFN-alpha and IL-12 levels were remarkably lower in LILs than in PBMCs,but IL-10 was higher when stimulated with DC-specific stimulator,polyI:C,CpG2216 and IL-3;The mDCs and pDCs were found to be infiltrated in the livers of ACHBLF patients as compared with CHB or healthy subjects.Conclusions The intrahepatic DC subsets preferentially produces IL-10,which might exert an important role in immune regulation in situ of livers of ALHBLF patients.