自体树突状细胞来源的Exosome活化HBV携带者PBMCs的体外研究

目的探讨负载HBcAg的树突状细胞（DCs）来源的Exosomes（DEXs）体外活化自体外周血单个核细胞（PBMCs）的能力,并对慢性乙型肝炎病毒（HBV）感染者与自然免疫获得者来源的DEXs的功能进行比较。方法培养PBMCs来源的DCs,分级离心法收获DEXs,电镜及Western blot分析DEXs结构及蛋白成分,MTS和ELISPOT法检测负载HBcAg的DEXs体外活化自体PBMCs增殖及产IFN-γ的能力。结果负载HBcAg的成熟DEXs体外刺激自体PBMCs增殖能力强于未成熟者（P〈0.01）,自然免疫者与慢性感染者间差异无统计学意义;成熟DEXs刺激PBMC产IFN-γ能力亦强于未成熟者（182±7）vs（14±2）,P〈0.05],且自然免疫者刺激PBMCs产IFN-γ能力强于慢性感染者（68±12,P〈0.05）。慢性感染者mDCs联合mDEXs刺激PBMCs产IFN-γ能力强于单用mDCs（326±3）vs（275±23）,P〈0.05],相当于自然免疫获得者联合组（396±15,P〉0.05）。结论 mDEXs可有效活化自体PBMCs,且自然免疫者mDEXs活化PBMCs能力强于慢性感染者,这一现象可能与慢性感染者体内T细胞活化过程存在缺陷有关。

HBcAg-pulsed-dendritic cell-derived exosomes induce the cellular response of peripheral blood mononuclear cells in vitro

Objective To investigate the capacity of HBcAg-pulsed-dendritic cell-derived exosomes（DEXs） for inducing the cellular response of peripheral blood mononuclear cells in vitro.Methods The mononuclear cells derived dendritic cells（DCs） were collected and generated from chronic HBV（CHB） patients and HBV naturally immunized healthy donors.DEXs were purified by ultracentrifugation and identified by transmission electron microscope and Western blotting.The assays of autologous lymphocyte proliferation and ELISPOT were used to evaluate the efficacy of HBcAg-pulsed DEXs.Results HBcAg-pulsed-dendritic cell mature DEXs induced stronger PBMCs proliferation than immature DEXs didA：（0.90±0.04）vs（0.47±0.02）,P0.001].However,the ability of PBMCs proliferation induction was weaker for mature DEXs than for mature DCs（P0.05）.HBcAg-pulsed-dendritic cell mature DEXs induced stronger IFN-γ production of PBMCs than immature DEXs did（182±7） vs（14±2）,P0.05].The mature DEXs from CHB patients were weaker in inducing the IFN-γ production of PBMCs than the mature DEXs from naturally immunized healthy donors.Conclusions HBcAg-pulsed-dendritic cell mature DEXs could stimulate potent PBMCs proliferation and IFN-γ production.The DEXs from CHB patients showed decreased PBMCs stimulatory ability,which might be partially related to HBV disease progression.