表达gH/gL抗原的腺病毒载体EBV疫苗免疫BALB/c小鼠后的T细胞表位鉴定  

Identification of the T-cell Epitope in BALB/c Mice Immunized with Adenovirus Vector EBV Vaccine Expressing gH/gL Antigen

作　　者：石雪洋 郝彦哲[2] 李晨雨 马琦 王冰丹 冯霞[2] 李淑英 SHI Xueyang;HAO Yanzhe;LI Chenyu;MA Qi;WANG Bingdan;FENG Xia;LI Shuying(Hebei Key Laboratory for Chronic Diseases,North China University of Science and Technology Basic Medical College,Tangshan 063000,China;National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases,National Institute for Viral Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Bejing 100052,China;North China University of Science and Technology Basic Medical College,Tangshan 063000,China)

机构地区：[1]华北理工大学基础医学院,河北省慢性疾病基础医学重点实验室,唐山063000 [2]传染病溯源预警与智能决策全国重点实验室,中国疾病预防控制中心病毒病预防控制所,北京100052 [3]华北理工大学公共卫生学院,唐山063000

出　　处：《病毒学报》2024年第3期500-507,共8页Chinese Journal of Virology

基　　金：艾滋病和病毒性肝炎等重大传染病防治科技重大专项(项目号:2018ZX10731101-002-010),题目:MVA及Ad5载体疫苗联合应用的临床前研究;河北省自然基金生物医药联合基金培育项目(项目号:H2022209049),题目:HPV E6E7融合基因重组病毒的构建及其抗肿瘤机制研究。

摘　　要：为了鉴定表达爱泼斯坦-巴尔病毒(Epstein-Barr virus,EBV)gH/gL保护性抗原的腺病毒载体疫苗在BALB/c小鼠中的T细胞表位,本研究使用表达gH/gL蛋白的腺病毒载体EBV疫苗免疫BALB/c小鼠,利用IEDB Analysis Resource软件对序列进行亲和力预测,设计合成EBV gH基因和gL基因的肽库,并将其混合成肽池,取免疫后的小鼠脾细胞进行IFN-γ ELISPOT检测,通过肽池和单肽两轮筛选检测具有阳性反应的多肽片段,确定优势反应肽。结果显示筛选出gH基因5条特异性优势肽,其中序列为LRGPFSYPSLTSAQS的单肽反应最强;gL基因4条特异性优势肽,其中序列为ASLNSPKNGSNQLVI的单肽反应最强。本研究使用IFN-γ ELISPOT法筛选和确定了9条EBV gH抗原特异性和gL抗原特异性优势T细胞表位,为EBV免疫学与疫苗研究提供了参考。We wished to identify the T-cell epitopes of an adenovirus vector vaccine expressing the protective antigen of gH/gL of the Epstein–Barr virus(EBV)in BALB/c mice.We immunized BALB/c mice with an adenovirus vector EBV vaccine expressing gH/gL protein.We predicted the affinity of the sequence with Immune Epitope Database Analysis Resource software.Next,we designed and synthesized the peptide library of gH and gL of EBV,and mixed them into a“peptide pool”.Interferon(IFN)-γin immunized mouse spleen cells was detected using enzyme-linked immunosorbent spot(ELISpot).Peptide fragments with a positive reaction were detected by two rounds of peptide-pool and single-peptide screening to determine the dominant reactive peptide.Five specific dominant peptides of gH were screened out,among which the single peptide with the sequence of LRGPFSYPSLTSAQS had the strongest reaction.There were four specific dominant peptides in gL,among which the single peptide with the sequence ASLNSPKNGSNQLVI had the strongest reaction.In summary,an ELISpot method based on IFN-γwas used to screen and determine nine dominant T-cell epitopes specific to the gH antigen and gL antigen of EBV.Our method provides a reference for EBV immunology and vaccine research.

关 键 词：EBV抗原 T细胞表位 IFN-γELISPOT 

分 类 号：R392.9[医药卫生—免疫学]