HBsAg和HBcAg联合负载DC与CIK共培养细胞体系的建立和功能评估  

Establishment and functional evaluation of DC-CIK co-culture system with combined loading of HBsAg and HBcAg

作　　者：王少扬[1] 林涛发 朱玲玲[1] 谢丽平[1] WANG Shaoyang LIN Taofa ZHU Lingling et al.(Department of Infectious Diseases, Fuzhou General Hospital of Nanjing Military Area Command, Fuzhou 350003, Chin)

机构地区：[1]南京军区福州总医院感染科厦门大学附属东方医院感染病学教研室,福州350003

出　　处：《临床肝胆病杂志》2017年第7期1280-1283,共4页Journal of Clinical Hepatology

基　　金：福建省科技计划项目重点项目(2013Y0073)

摘　　要：目的观察在不同HBV抗原负载下诱导的树突状细胞(DC)同细胞因子诱导的杀伤细胞(CIK)共培养后CIK的功能。方法将2015年1月-2016年6月在南京军区福州总医院就诊的13例慢性乙型肝炎患者作为研究对象,分离13例慢性乙型肝炎患者的外周血单个核细胞,并培养DC和CIK,设为CIK单独培养组、DC+CIK共培养组、DC+CIK+HBsAg共培养组、DC+CIK+HBcAg共培养组、DC+CIK+HBsAg+HBcAg共培养组,培养完成后,用ELISPOT方法测定CIK产生IFNγ的能力,用HepG2.2.15细胞作为靶细胞,测定CIK的杀伤功能。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果不同CIK试验组产IFNγ的水平差异有统计学意义(F=29.84,P<0.001),其中以DC+CIK+HBsAg+HBcAg组产生的IFNγ水平最高;CIK杀伤率组间比较差异有统计学意义(F=14.77,P<0.001),其中以DC+CIK+HBsAg+HBcAg最为突出。结论 HBsAg和HBcAg联合负载的DC与CIK共培养可能是治疗慢性乙型肝炎有效的细胞体系。Objective To investigate the function of cytokine-induced killer cell（CIK） after co-culture of dendritic cells（DC） and CIK loaded with different HBV antigens.Methods A total of 13 patients with chronic hepatitis B（CHB） who visited Fuzhou General Hospital of Nanjing Military Area Command from January 2015 to June 2016 were enrolled.Peripheral blood mononuclear cells（PBMCs） were isolated from 13 CHB patients,and DC and CIK were cultured and divided into CIK culture group,DC＋CIK co-culture group,DC＋CIK ＋HBsAg co-culture group,DC＋CIK＋HBcAg co-culture group,and DC＋CIK＋HBsAg＋HBcAg co-culture group.Then ELISPOT assay was performed to measure the ability of CIK to produce pegylated interferonγ（IFNγ）,and HepG2.2.15 cells were used as target cells to measure the killing function of CIK.A one-way analysis of variance was used for comparison of continuous date between groups,and the least significant difference t-test was used for further comparison between any two groups.Results There was a significant difference in IFNγ level between different CIK test groups（F=29.84,P〈0.001）,and the DC＋CIK＋HBsAg＋HBcAg group had the highest IFNγlevel.There was a significant difference in CIK killing rate between groups（F=14.77,P〈0.001）,and the DC＋CIK＋HBsAg＋HBcAg group had the highest CIK killing rate.Conclusion DC-CIK co-culture with combined loading of HBsAg and HBcAg may be an effective cell system for the treatment of CHB.

关 键 词：肝炎 乙型 慢性 肝炎表面抗原 乙型 肝炎核心抗原 乙型 树突细胞 杀伤细胞 

分 类 号：R512.62[医药卫生—内科学]