乙肝病毒表面抗原冲击的树突状细胞诱导记忆T细胞的形成及增殖研究  

Formation and proliferation of memory T cells induced by hepatitis B virus surface antigen-pulsed dendritic cells

作　　者：黄新亮 余方流 伍云云 王凤各 董博翰 Huang Xinliang;Yu Fangliu;Wu Yunyun;Wang Fengge;Dong Bohan(School of Basic Medicine,Wannan Medical College,Wuhu 241002,Anhui,China)

机构地区：[1]皖南医学院基础医学院,安徽芜湖241002

出　　处：《右江民族医学院学报》2021年第3期339-343,共5页Journal of Youjiang Medical University for Nationalities

基　　金：2019年度安徽高校省级自然科学研究重点项目(KJ2019A0415)。

摘　　要：目的探索在体外用乙肝病毒表面抗原(HBsAg)冲击的树突状细胞(DC)诱导记忆T细胞(Tm)的形成及增殖。方法选择健康志愿者,分低剂量实验组、中剂量实验组、高剂量实验组各6例,且分相应的低剂量自身对照组、中剂量自身对照组、高剂量自身对照组各6例。低剂量实验组、中剂量实验组、高剂量实验组分别以10 ng/ml、20 ng/ml、30 ng/ml HBsAg冲击DC;低剂量自身对照组、中剂量自身对照组、高剂量自身对照组均不用HBsAg冲击DC。培养第8天流式仪鉴定DC。后与自身淋巴细胞共培养7天后用流式细胞仪检测CD45RO+CD4+Tm百分率。再以MTT法检测CD45RO+CD4+Tm的增殖活性。结果低剂量实验组、中剂量实验组、高剂量实验组分别比其相应自身对照组DC中表达CD80、CD40、HLA-DR的百分率、CD45RO+CD4+Tm的百分率、CD45RO+CD4+Tm的增殖活性显著升高(P<0.05)。且中剂量实验组、高剂量实验组分别比低剂量实验组的CD45RO+CD4+Tm增殖活性更高,差异具有统计学意义(P<0.05)。结论体外用HBsAg冲击的DC能够诱导CD45RO+CD4+Tm的形成及增殖,且用20 ng/ml、30 ng/ml HBsAg分别比10 ng/ml HBsAg诱导的CD45RO+CD4+Tm的增殖活性更高,这为探索免疫记忆机制奠定了依据。Objective To explore the formation and proliferation of memory T(Tm)cells induced by dendritic cells(DCs)which were pulsed with hepatitis B surface antigen(HBsAg)in vitro.Methods Healthy volunteers were divided into the low-dose,medium-dose and high-dose experimental groups with 6 cases in each group and corresponding low-dose,medium-dose and high-dose self control groups with 6 cases in each group.DCs in the low-dose,medium-dose and high-dose experiments groups were pulsed with 10 ng/ml,20 ng/ml and 30 ng/ml HBsAg,respectively.The low-dose,medium-dose and high-dose self control groupal were not pulsed with HBsAg.The DCs were identified by flow cytometry on the eighth day of culture.The percentage of CD45RO+CD4+Tm was detected by flow cytometry after 7 days of co-culture with autolymphocytes.The proliferation activity of CD45RO+CD4+Tm was detected by MTT assay.Results The percentage of CD80,CD40 and HLA-DR expressed in DC,the percentage of CD80,CD40,HLA-DR,the percentage of CD45RO+CD4+Tm and the proliferation activity of CD45RO+CD4+Tm in the low-dose,medium-dose and high-dose experimental groups were significantly higher than those in the corresponding self control groups(P<0.05).Moreover,the proliferation activity of CD45RO+CD4+Tm in the medium-dose and high-dose experimental groups was higher than that in the low-dose experimental group,and the difference was statistically significant(P<0.05).Conclusion DCs pulsed with HBsAg can induce the formation and proliferation of CD45RO+CD4+Tm in vitro,and the proliferation activity of CD45RO+CD4+Tm induced by 20 ng/ml and 30 ng/ml HBsAg is higher than that induced by 10 ng/ml HBsAg,respectively.This provides a basis for exploring the mechanism of immune memory.

关 键 词：树突状细胞 乙型肝炎病毒表面抗原 记忆T细胞 免疫 

分 类 号：R392[医药卫生—免疫学]