HLA-DRB1 ＊0803与HLA-DRB1 ＊0901等位特异性HBcAg表位肽的筛查与鉴定  

Screening and identification of HLA-DRB1*0803 and HLA-DRB1*0901 restricted epitopes from hepatitis B core antigen

作　　者：李辉文[1] 何韦韦 谭文婷[1] 王莉 陈晓玲 邓国宏[1] 

机构地区：[1]第三军医大学、西南医院全军感染病研究所、感染病研究重庆市重点实验室,重庆400038 [2]全军免疫学研究所,重庆400038

出　　处：《第三军医大学学报》2015年第9期833-838,共6页Journal of Third Military Medical University

基　　金：国家自然科学基金重点项目(81330038);“十二五”传染病国家科技重大专项子课题(2012ZX10002007-002-005);第三军医大学临床科研重大专项(2012XLC05)~~

摘　　要：目的鉴定HLA-DRB1*0803与HLA-DRB1*0901等位特异性HBV表位,进一步认识HLA-DR不同等位限制性表位的差异。方法 BLCL细胞表面高表达HLA-DR分子,从HBc Ag肽池负载的特征HLA-DR等位的BLCL细胞表面上收集相关多肽,多肽经过过滤、脱盐、浓缩等处理步骤后对其进行液质联用质谱分析(LC-MS),再结合生物信息学预测工具Net MHCⅡpan 3.0 Server初步确定其潜在候选表位。结果筛选出HLA-DRB1*0803限制性表位HBc17-31、HBc22-36、HBc27-41、HBc82-96、HBc87-101、HBc92-106、HBc117-131、HBc122-136共8条,HLA-DRB1*0901限制性表位HBc2-16、HBc7-21、HBc12-26、HBc27-41、HBc82-96、HBc87-101、HBc122-136共7条,其中相同的有4条。结论发现了HBc7-21、HBc22-36、HBc27-41、HBc82-96、HBc87-101、HBc92-106新的HLA-Ⅱ类分子表位,不同的HLA-DR等位限制的HBc Ag表位肽存在差异。Objective To identify the specific epitopes of HLA-DRB1 ＊0803 and HLA-DRB1 ＊ 0901 alleles from hepatitis B virus for further understanding the difference of the epitopes restricted to the different HLA-DR alleles. Methods After incubating the peptide pool of pre-core and core sequence of hepatitis B virus with a B lymphoblastoid cell fine （BLCL, holnozygous human B cell lines of allele HLA-DRB1 ＊0803 or HLA-DRB1 ＊ 0901 with cell surface highly expressing HLA-DR molecules）, the peptides were eluted and collected from the BLCL surface without affecting cell viability. Then the peptides were filtered, desalted, concentrated and analyzed by liquid chromatography-mass spectrometry （LC-MS）. Bioinformatics predictive tool NetMHC Ⅱ pan 3.0 Server was employed to confirm the candidate epitopes. Results There were 8 potential epitopes restricted to HLA-DRB1 ＊ 0803 finally identify from HBV, that is, HBcl7-31, HBc22-36, HBc27-41, HBc82-96, HBc87-101, HBc92-106, HBcll7-131, and HBc122-136, and 7 potential epitopes restricted to HLA-DRB1 ＊ 0901, that is, HBc2-16, HBc7-21, HBc12-26, HBc27-41, HBc82-96, HBc87- 101, and HBc122-136. Four of them were same epitopes. Conclusion We have found 6 new epitopes to HLA class II , ie, HBc7-21, HBc22-36, HBc27-41, HBc82-96, HBc87-101 and HBc92-106, and there are some differences among the HBcAg epitopes restricted to different HLA-DR alleles.

关 键 词：HLA-DRB1＊0803 HLA-DRB1＊0901 BLCL HBCAG 质谱 

分 类 号：R341[医药卫生—基础医学] R392.3