>
网站首页期刊介绍通知公告编 委 会投稿须知电子期刊广告合作联系我们
最新消息:
HLA-A0201、A1101和A2402限制性HBV抗原肽的虚拟预测与实验鉴定
作者:赵晨1  金萧萧1  丁艳1  昂倩倩1  Odontuya Khaidav1  夏玲芝2  邱洁3  沈传来1 
单位:1. 东南大学医学院 病原生物学与免疫学系, 江苏 南京 210009;
2. 南京金域医学检验所 自身免疫病检测中心, 江苏 南京 210032;
3. 南京市第二医院 传染病科, 江苏 南京 210003
关键词:人类白细胞抗原-A 乙型肝炎病毒 T细胞表位肽 γ干扰素酶联免疫斑点法 
分类号:R373.21;R392.1
出版年·卷·期(页码):2019·38·第六期(989-996)
摘要:

目的:筛选和鉴定基因总频率> 50%的3种人类白细胞抗原(HLA)(A0201、A1101和A2402)分子限制的乙型肝炎病毒(HBV)抗原T细胞表位肽。方法:利用6种在线T细胞表位肽预测数据库,针对乙肝病毒表面抗原、核心抗原、DNA多聚酶和X蛋白等4种抗原,虚拟筛选3种HLA-A分子限制的T细胞表位肽。从南京市第二医院检验科收集乙型肝炎住院患者的外周抗凝血,制备外周血单个核细胞,通过γ干扰素酶联免疫斑点法(IFN-γ ELISPOT)筛选出对任何一组混合多肽呈现特异T细胞反应的乙型肝炎患者;重新采集这些乙型肝炎患者的新鲜外周血,用IFN-γ ELISPOT法鉴定混合肽中单种抗原肽的免疫原性;并采用聚合酶链反应-测序分型法对这些乙型肝炎患者进行HLA-A等位基因分型。结果:经鉴定具有免疫原性的HLA-A0201分子限制性HBV表位肽8种,A1101限制性HBV表位肽6种,A2402限制性HBV表位肽7种。结论:本研究筛选鉴定了3种HLA-A分子限制的4种HBV抗原的21种T细胞表位肽,其中11种未见报道,而且HLA-A1101分子限制性表位肽的免疫原性明显弱于HLA-A0201和A2402分子限制性表位肽。

Objective: To screen and identify the hepatitis B virus (HBV) antigenic T cell epitopes restricted by human leukocyte antigen (HLA)-A0201, A1101 and A2402 which have a total gene frequency of >50% in Chinese population. Methods: Six online data banks for T cell epitopes prediction were used to virtually screen the T cell epitopes derived from HBsAg, HBcAg, HBpol and HBx proteins and restricted by the three HLA-A molecules,respectively. Peripheral anticoagulation samples from HBV-infected in-patients were collected from the Department of Clinical Laboratory of Nanjing the Second Hospital, and peripheral blood mononuclear cells (PBMCs) were processed.Then the interferon-γ enzyme linked immunospot (IFN-γ-ELISPOT) assay was performed to screen out the patients who had the specific T cell responses to the cocktail of peptides. The fresh peripheral blood samples were collected again from these patients and the immunogenicity of each peptide was confirmed by the IFN-γ-ELISPOT assay. Meanwhile, HLA-A genotyping was carried out in these HBV-infected patients through polymerase chain reaction-sequencing-based typing (PCR-SBT) method. Results: The immunogenicity of eight epitopes restricted by HLA-A0201, six epitopes restricted by A1101, and seven epitopes restricted by A2402 were confirmed by functional experiment. Conclusion: Twenty-one T cell epitopes have been identified, which derived from four kinds of HBV proteins and restricted by three HLA-A molecules, respectively. Of them, eleven epitopes have not been reported before. Moreover, the immunogenicity of the epitopes restricted by HLA-A1101 is significantly weaker than that restricted by HLA-A0201 and A2402.

参考文献:

[1] 周光炎.免疫学原理[M].4版.北京:科学出版社,2018.
[2] ELAHI S,HORTON H.Association of HLA-alleles with the immune regulation of chronic viral infections[J].Int J Biochem Cell Biol,2012,44(8):1361-1365.
[3] 李兰娟,王宇明.感染病学[M].3版.北京:人民卫生出版社,2015.
[4] AYALEW M B,HORSSA B A,GETACHEW N,et al.Knowledge and attitude of health care professionals regarding hepatitis B virus infection and its vaccination,University of Gondar Hospital,Ethiopia[J].Hepat Med,2016,8:135-142.
[5] TSENG T C,HUANG L R.Immunopathogenesis of hepatitis B virus[J].J Infect Dis,2017,216(Suppl_8):S765-S770.
[6] WANG Q,PAN W,LIU Y,et al.Hepatitis B virus-specific CD8+T cells maintain functional exhaustion after antigen reexposure in an acute activation immune environment[J].Front Immunol,2018,9:219.
[7] PAN N,CHEN K,QIU J,et al.Human leukocyte antigen class I alleles and haplotypes associated with primary hepatocellular carcinoma in persistent HBV-infected patients[J].Hum Immunol,2013,74(6):758-763.
[8] LIU Q,ZHENG Y,YU Y,et al.Identification of HLA-A*0201-restricted CD8+T-cell epitope C64-72 from hepatitis B virus core protein[J].Int Immunopharmacol,2012,13(2):141-147.
[9] TAN A T,SODSAI P,CHIA A,et al.Immunoprevalence and immunodominance of HLA-Cw*0801-restricted T cell response targeting the hepatitis B virus envelope transmembrane region[J].J Virol,2014,88(2):1332-1341.
[10] YAMAMIYA D,MIZUKOSHI E,KAJI K,et al.Immune responses of human T lymphocytes to novel hepatitis B virus-derived peptides[J].PLoS One,2018,13(6):e0198264.
[11] 李杨,何军,鲍晓晶,等.中国汉族人群供-受者HLA-A、B、Cw、DRB1和DQB1高分辨等位基因频率分布及错配比例的研究[J].中华医学遗传学杂志,2011,28(1):92-98.
[12] 黄庆海,吴亚萌,李震,等.基于HLA等位基因频率的全球人群亲缘关系初探[J].东南大学学报(医学版),2014,33(4):429-435.
[13] VOORTER C E,PALUSCI F,TILANUS M G.Sequence-based typing of HLA:an improved group-specific full-length gene sequencing approach[J].Methods Mol Biol,2014,1109:101-114.
[14] COMBER J D,KARABUDAK A,SHETTY V,et al.MHC class I presented T cell epitopes as potential antigens for therapeutic vaccine against HBV chronic infection[J].Hepat Res Treat,2014,2014:860562.
[15] LI J,HAN Y,JIN K,et al.Dynamic changes of cytotoxic T lymphocytes (CTLs),natural killer (NK) cells,and natural killer T (NKT) cells in patients with acute hepatitis B infection[J].Virol J,2011,8:199.

服务与反馈:
文章下载】【发表评论】【查看评论】【加入收藏
提示:您还未登录,请登录!点此登录
您是第 232892 位访问者


copyright ©《东南大学学报(医学版)》编辑部
联系电话:025-87232481 83272483
电子邮件:
bjb@pub.seu.edu.cn

苏ICP备09058364