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.
作者
黄杰庭
尤庆柱
李丰沛
许茹
王敏
廖峭
单振刚
戎霞
付涌水
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)
出处
《中国输血杂志》
CAS
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)。
