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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。
Abstract：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)-γ in 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-γ was 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.
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基本信息:
DOI：10.13242/j.cnki.bingduxuebao.004490
中图分类号:R392
引用信息:
[1]石雪洋,郝彦哲,李晨雨等.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[J].病毒学报,2024,40(03):500-507.DOI:10.13242/j.cnki.bingduxuebao.004490.
基金信息:
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~~