不同阶段 HBV 相关 ACLF 患者外周血 DC 及T 淋巴细胞免疫功能特点研究

目的：比较不同阶段的 HBV 相关慢加急性（亚急性）肝衰竭（HBV-ACLF）患者外周血树突状细胞（DC）、T淋巴细胞（TC）相关细胞免疫功能，阐述 DC-TC 轴在 HBV-ACLF 发病过程中可能的细胞免疫学机制。方法HBV-ACLF患者30例，分为早期组15例与中晚期组15例，另设健康对照组8例，以外周血来源的 PBMC 体外分离诱导培养 DC 与TC，应用流式细胞计数检测 DC 细胞表型 HLA-DR、CD80、CD86、CD83、CD1α的表达率，及 TC 表面分子 CD3＋、CD4＋ T、CD8＋ T 淋巴细胞百分比，并检测 DC 上清液中 IFN-α、IL-4的分泌水平，比较不同阶段 HBV-ACLF 患者免疫细胞及炎性因子表达的差异。结果与健康人比较，HBV-CLF 患者 DC 表型 HLA-DR、CD1α、CD83、CD80、CD86表达率显著下降（t 值分别为5．3356、13．269、10．8742、13．3685和23．021，均 P ＜0．01），DC 分泌因子 IFN-α显著升高（t 值为16．4569，P ＜0．01）；TC 表面分子 CD3＋、CD4＋ T、CD4＋／CD8＋细胞比值显著下降（t 值分别为7．4441、12．5557、11．0771，均 P ＜0．01）， CD8＋ T 细胞百分比显著上升（t＝4．4359，P ＜0．01）；HBV-ACLF 患者中晚期组 DC 表型 CD83、CD86表达率显著低于早期组（P 值分别为：0．0000，0．0057），DC 分泌因子 IFN-α表达在早期组显著增多（P ＝0．0000），IL-4表达在中晚期组显著增多（P ＝0．0000），TC 表面分子中晚期组 CD4＋ T 细胞百分比、CD4＋／ CD8＋细 胞 比 值 显 著 下 降 （P 值分别为：0．0268、0．0002），CD8＋ T 细胞百分比显著上升（P ＝0．0001）。结论不同阶段 HBV-ACLF 患者的 DC、TC 功能状态均表现为细胞免疫功能低下，中晚期患者的细胞免疫功能更为低下；HBV-ACLF 全病程存在促／抑炎性细胞因子功能紊乱，早期患者存在炎症因子过度释放，中晚期患者存在抗炎症细胞因子表达增强。

Investigation on immunologic function of dendritic cells and T lymphocytes in different stages of hepatitis B virus related acute on chronic liver failure patients

Objective To compare the function of dendritic cells （DCs）and T lymphocytes （TCs）in different stages of hepatitis B virus related acute on chronic liver failure （HBV-ACLF）patients,and elaborate the potential roles of DCs-TCs in cell immunological mechanism of the pathogenesis of HBV-ACLF.Methods HBV-ACLF patients were divided into two groups,including early stage group,and middle and advanced stage group.A healthy control group was set as well. DCs and TCs were separated and cultured in vitro from peripheral blood mononuclear cells （PBMCs）in peripheral blood. Flow cytometry was used to detect the expressions of surface molecules of DCs including HLA-DR,CD80,CD86,CD83, CD1α,and surface molecules of TCs including CD3 ＋,CD4＋,CD8 ＋;IFN-α,and IL-4 levels in supernatant of DCs,to demonstrate whether there were differences in inflammatory cytokines and immune cells expressions between different stages of HBV-ACLF.Results Compared with those in healthy people,expression rates of DCs phenotype HLA-DR,CD1α, CD83,CD80 and CD86 in patients with HBV-ACLF were significantly decreased（t =5.3356,13.269,10.8742,13.3685and 23.021 ,respectively,P 〈0.01）,while excreted factor IFN-αhad a significant rise（t =16.4569,P 〈0.01 ）;expression rates of TCs phenotype CD3 ＋,CD4 ＋ and CD4 ＋/CD8 ＋ were significantly decreased（t =7.4441 ,12.5557,11 .0771 , respectively,P 〈0.01 ）,while percentage of TCs phenotype CD8 ＋ was significantly increased（t =4.4359,P 〈0.01 ）. Compared with early stage group,middle and advanced stage group showed significantly lower expression rates of CD83 and CD86（P =0.0000,0.0057）,as well as lower expression rates of TCs phenotype CD4＋ and CD4＋/ CD8 ＋（P =0.0268, 0.0002）.There was a significantly higher level of IFN-α in early stage （P =0.0000）than that in middle and advanced stage,while IL-4 showed opposite（P =0.0000）.Conclusion DCs and TCs in patients with HBV-ACLF in different stages all showed a low immune function,especially in the mid