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

王少扬; 林涛发; 朱玲玲; 谢丽平

doi:10.3969/j.issn.1001-5256.2017.07.015

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

DOI: 10.3969/j.issn.1001-5256.2017.07.015

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

基金项目:

福建省科技计划项目重点项目(2013Y0073)；

详细信息

中图分类号: R512.62
计量
- 文章访问数: 2257
- HTML全文浏览量: 34
- PDF下载量: 406
- 被引次数: 0
出版历程
- 收稿日期: 2017-01-24
- 出版日期: 2017-07-20

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

Department of Infectious Diseases, Fuzhou General Hospital of Nanjing Military Area Command

Research funding:

摘要

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...
- 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
Abstract: 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.
- hepatitis B, chronic /
- hepatitis B surface antigen s /
- hepatitis B core antigens /
- dendritic cells /
- kill cells

HTML全文

参考文献(11)

[1]WANG RB, HAO DY, ZOU JY, et al.Influence of DC-CIK immunotherapy in blood coagulation function and D-dimer levels in patients with ovarian cancer[J].J Jilin Univ:Med Edit, 2017, 43 (1) :120-124. (in Chinese) 王睿斌, 赫东芸, 邹积艳, 等.DC-CIK免疫治疗对卵巢癌患者凝血功能及D-二聚体水平的影响[J].吉林大学学报:医学版, 2017, 43 (1) :120-124.

[2] Chinese Society of Hepatology, Chinese Society of Infectious Diseases, Chinese Medical Association.The guideline of prevention and treatment for chronic hepatitis B (2010 version) [J].J Clin Hepatol, 2011, 27 (1) :Ⅰ-ⅩⅥ. (in Chinese) 中华医学会肝病学分会, 中华医学会感染病学分会.慢性乙型肝炎防治指南 (2010版) [J].临床肝胆病杂志, 2011, 27 (1) :Ⅰ-ⅩⅥ.

[3]AKBAR SM, HORIIKE N, ONJI M.Immune therapy including dendriticcell based threapy in chronic hepatitis B vrirus infection[J].World J Gastronterol, 2006, 12 (18) :2876-2883.

[4]VAN RG, SPRENGERS D, BINDA RS, et al.Funetional impairment of myeloid and plasmacytoid dendritie cells of patienis with chronic hepatitis B[J].Hepatology, 2004, 40 (3) :738-746.

[5]ZHENG BJ, ZHOU J, QU D, et al.Selective functional deficit in dendritic cell-T interaction is a crucial mechanism in chronic hepatitis B virus infection[J].J Viral Hepat, 2004, 11 (3) :217-224.

[6]TAVAKOLI S, SCHWERIN W, ROHWER A, et al.Phenotype and function of monocyte derived dendritic cells in chronic hepatitis B virus infection[J].J Gen Virol, 2004, 85 (Pt 10) :2829-2836.

[7]WANG SY, LIN TF, LIU HZ, et al.A clinical study of autologous cytokine-induced killer cells in treatment of patients with chronic hepatitis B[J].J Clin Hepatol, 2015, 31 (4) :530-533. (in Chinese) 王少扬, 林涛发, 刘海周, 等.自体细胞因子诱导的杀伤细胞治疗慢性乙型肝炎患者的临床观察[J].临床肝胆病杂志, 2015, 31 (4) :530-533.

[8]LIU YZ, LUO Y, XIE SQ, et al.Progress of the Anti-tumor effects and mechanism of DC-CIK[J].Anti-tumor Pharmacy, 2016, 6 (2) :108-113. (in Chinese) 刘垠子, 罗以, 谢思琪, 等.DC-CIK的抗肿瘤作用及机制的研究进展[J].肿瘤药学, 2016, 6 (2) :108-111.

[9]SHI M, QIAN S, CHEN WW, et al.Hepatitis B virus (HBV) antigen-pulsed monocyte-derived dendritic cells from HBV-associated hepatocellular carcinoma patients significantly enhance specific T cell responses in vitro[J].Clin Exp Immunol, 2014, 147 (2) :277-286.

[10]DUAN XZ, HE HX, ZHUANG H.Restoration in vitro of impaired T-cell responses in patients with chronic hepatitis B by autologous dendritic cells loaded with hepatitis B virus proteins[J].J Gastrol Hepatol, 2006, 21 (6) :970-976.

[11]ZHAO X, CHANG Y, LIU YX, et al.Experiment study of killing effect of DC and CIK co-cultured on tumor cells[J].Chin J Lab Diag, 2015, 19 (10) :1636-1638. (in Chinese) 赵鑫, 常颖, 刘玉侠, 等.DC、CIK联合培养对肿瘤细胞杀伤作用的实验研究[J].中国实验诊断学, 2015, 19 (10) :1636-1638.

施引文献

资源附件(0)

访问统计