CD5^+CD19^+B细胞在乙型肝炎病毒慢性感染中对CD8+细胞的作用和机制  

Effects and mechanisms of CD5^+ CD19^+ B lymphocytes on CD8+ cells in chronic hepatitis B virus infection

作　　者：周震宇 霍佳慧 龚镭 胡臻 唐栓 孟淼 蒋文瑜 凤辉 王小云 Zhou Zhenyu;Huo Jiahui;Gong Lei;Hu Zhen;Tang Shuan;Meng Miao;Jiang Wenyu;Feng Hui;Wang Xiaoyun(Department of Gastroenterology,Wuxi Second People′s Hospital Affiliated to Nanjing Medical University,Wuxi 214000,China;Department of Gastroenterology,People′s Hospital of Yixing)

机构地区：[1]南京医科大学附属无锡第二医院消化内科,无锡214000 [2]宜兴市人民医院消化内科,214200

出　　处：《中华消化杂志》2020年第9期617-623,共7页Chinese Journal of Digestion

基　　金：国家自然科学基金(81500467);江苏省自然科学基金(BK20151116);无锡市医学发展学科建设对象(ZDXK002);无锡市医院管理中心科研项目(YGZXZ1504);无锡市第二人民医院樊代明院士工作站(CYR1705);江苏省青年医学重点人才(QNRC2016151)。

摘　　要：目的观察CD5^+CD19^+B细胞在体外是否具有分泌IL-10的功能,并探讨其在HBV感染过程中对CD8^+细胞的作用和机制。方法纳入2017年7月至2018年6月在南京医科大学附属无锡第二医院确诊的慢性乙型肝炎(乙肝组)23例、肝硬化(肝硬化组)18例患者,以及同期健康对照(健康对照组)19名。通过外周血单个核细胞(PBMC)分离、细胞培养、流式细胞术分析、CD5^+CD19^+B细胞分离等操作,比较3组CD5^+CD19^+B细胞含量较多(占淋巴细胞百分比>6%)的患者比例、CD5^+CD19^+B细胞分泌IL-10的情况和IL-10^+细胞含量较多(占淋巴细胞百分比>4%)的患者比例,分析CD5^+CD19^+B细胞对CD8^+细胞分泌γ干扰素的影响和可能机制。通过对慢性乙型肝炎13例和肝硬化5例患者行肝活组织和免疫组织化学检查,分析CD5^+CD19^+B细胞在人体肝组织中表达情况。统计学分析采用卡方检验和Fisher确切概率法。结果肝硬化组中CD5^+CD19^+B细胞含量较多的患者比例高于健康对照组(8/18比2/19),差异有统计学意义(Fisher确切概率法,P=0.029)。健康对照组(10名)、乙肝组(23例)、肝硬化组(18例)的CD5^+CD19^+B细胞分别占IL-10^+细胞的81.6%、82.3%、70.1%,IL-10^+细胞含量较多的患者分别为2、7和2例,3组比较差异均无统计学意义(P均>0.05)。在脂多糖刺激培养48 h后,健康对照组(10名)CD8^+γ干扰素^+细胞占淋巴细胞百分比与乙肝组(10例)、肝硬化组(10例)比较(10.73%比7.05%和9.52%)差异均无统计学意义(P均>0.05);剔除CD5^+CD19^+B细胞后,健康对照组(10名)、乙肝组(10例)和肝硬化组(10例)中分别有5、4、4例患者CD8^+γ干扰素^+细胞占淋巴细胞百分比升高。添加IL-10受体阻滞剂后,PBMC中CD8^+γ干扰素^+占淋巴细胞百分比较添加IL-10受体阻滞剂前升高(7.23%比6.87%)。肝活组织标本免疫组织化学分析结果显示,CD4^+和CD8^+细胞在患者肝小叶汇管区强表达,CD5^+和CD19^+细胞较少表达。结论CD5^+CDObjective To explore whether CD5^+CD19^+B cells has the function of secreteing interleukin-10(IL-10)in vitro,and to further investigate its possible effects and mechanisms on CD8^+cells in the process of hepatitis B virus(HBV)infection.Methods From July 2017 to June 2018,at Wuxi Second People′s Hospital Affiliated to Nanjing Medical University,23 patients with chronic hepatitis B(chronic hepatitis B group),18 patients with liver cirrhosis(liver cirrhosis group)and 19 healthy individuals in the same period as healthy controls(healthy control group)were enrolled.Peripheral blood mononuclear cell(PBMC)were isolated and cultured.CD5^+CD19^+B cells were isolated.The cells were analyzed by flow cytometry.The ratio of high CD5^+CD19^+B cells content(>6%of lymphocytes),the secretion of IL-10 by CD5^+CD19^+B and the ratio of high IL-10^+cells content(>4%of lymphocytes)of three groups were compared.The effects and possible mechanisms of CD5^+CD19^+B cells on the secreting of interferon-γ(IFN-γ)by CD8^+cells were analyzed.Liver biopsy and immunohistochemistry examination were conducted in 18 patients(13 patients with chronic hepatitis B and 5 patients with liver cirrhosis)and the expression of CD5^+CD19^+B cells in human liver tissues was analyzed.Chi square test and Fisher exact probability test were used for statistical analysis.Results The ratio of high CD5^+CD19^+B cells content of liver cirrhosis group was higher than that of healthy control group(8/18 vs.2/19)and the difference was statistically significant(Fisher exact probability test,P=0.029).The precentage of CD5^+CD19^+B cells in healthy control group(n=10),chronic hepatitis B group(n=23)and liver cirrhosis group(n=18)accounted for 81.6%,82.3%and 70.1%of IL-10^+cells,respectively,and the number of patients with high IL-10^+cells precentage was 2,7 and 2,respectively.There were no statistically significant differences among three groups(all P>0.05).After stimulated with lipopolysaccharide and cultured for 48 hours,the precentage of CD8^+IFN-γ^+cells in lympho

关 键 词：白细胞介素10 CD5^+CD19^+B细胞 CD8阳性T淋巴细胞 慢性乙型肝炎病毒感染 肝硬化 

分 类 号：R512.62[医药卫生—内科学]