基因1型和6型HCV CD4 T细胞抗原表位的初步鉴定  

Preliminary identification of CD4 T cell epitopes in genotype 1 and 6 of hepatitis C virus

作　　者：黄杰庭[1] 尤庆柱 李丰沛 许茹[1] 王敏[1] 廖峭[1] 单振刚[1] 戎霞[1] 付涌水[1] HUANG Jieting;YOU Qingzhu;LI Fengpei;XU Ru;WANG Min;LIAO Qiao;SHAN Zhengang;RONG Xia;FU Yongshui(Guangzhou Blood Center,Guangzhou Key Laboratory of Blood Safety,Guangzhou 510095,China;Department of Otolaryngology, Shaoguan First People′s Hospital)

机构地区：[1]广州血液中心广州市血液安全重点实验室,广州广东510095 [2]韶关市第一人民医院耳鼻喉科

出　　处：《中国输血杂志》2022年第7期693-697,共5页Chinese Journal of Blood Transfusion

基　　金：广东省医学科研基金(A2020199);国家自然科学基金面上项目(81772208);国家自然科学基金青年科学基金项目(32000666);广州市医学重点学科(2021-2023年)。

摘　　要：目的 研究基因1型(GT1)和6型(GT6)HCV Core蛋白和NS3蛋白的CD4 T细胞抗原表位。方法 合成覆盖GT1和GT6 HCV Core和NS3全长的重叠肽段(16-mer)298条。通过肽段库刺激17例HCV+献血者和7例健康献血者的外周血单个核细胞(PBMC),IFN-γELISPOT测定T细胞应答,初步筛选出21条阳性肽段。再将这些肽段单独刺激20例HCV+和18例健康献血者PBMC。比较2组之间对阳性肽段的应答频率的差异。结果 综合分析肽段库和单条肽段的试验结果,HCV+组对其中5条肽段(GT1 NS3_(1273-1288)和NS3_(1315-1330);GT6 NS3_(1033-1048)、NS3_(1087-1102)和NS3_(1351-1366))刺激的应答频率较高(18.9%~27.0%),且健康对照组无应答或低应答(0%~4.0%),2组之间应答频率有差异(P<0.05)。经序列比对,免疫表位数据库(IEDB)中没有与这5条肽段序列一致的抗原表位。结论 本研究鉴定出HCV GT1和GT6 HCV NS3蛋白新的CD4 T细胞抗原表位,对HCV疫苗的研发具有潜在的应用价值。Objective To study the CD4 T cell epitopes in Core and NS3 protein of genotype 1(GT1) and 6(GT6) of hepatitis C virus(HCV).Methods A total of 298 overlapping peptides(16-mer) spanning Core and NS3 protein of GT1 and GT6 HCV were synthesized.Peripheral blood mononuclear cells(PBMCs) from 17 HCV+ and 7 healthy blood donors were stimulated by peptide pools,followed by evaluating T cell response by IFN-γ ELISPOT,by which 21 peptides with positive results were found.These peptides were further applied to individually stimulate 20 HCV+ and 18 healthy PBMCs.The differences of responsive frequencies to the 21 positive peptides between the two study groups were compared.Results Pooled and individual peptide stimulation tests showed that HCV+ PBMCs were responsive to the stimulation of 5 peptides(GT1 NS3_(1273-1288) and NS3_(1315-1330);GT6 NS3_(1033-1048),NS3_(1087-1102) and NS3_(1351-1366)),with a responsive frequency ranging 18.9%-27.0%.In contrast,healthy PBMCs were not or low responsive(0%-4.0%) to these five peptides.The responsive frequencies were statistically different between the two groups(P<0.05).No reported epitopes in IEDB were found identical with these 5 peptides via sequence alignment.Conclusion Our study identified novel CD4 T cell epitopes in NS3 protein of GT1 and GT6 HCV,which has potential application value for the research and development of HCV vaccine.

关 键 词：丙型肝炎病毒 基因型 CD4 T细胞抗原表位 疫苗 献血者 

分 类 号：R457.1[医药卫生—治疗学] R373.2[医药卫生—临床医学]