目的:筛选柯萨奇病毒B3(Coxsackievirus B3,CVB3)中可被细胞毒性T细胞(cytotoxic T lymphocytes,CTL)高效识别的抗原表位,为开发针对CVB3及其他肠道病毒的广谱疫苗提供理论依据。方法:基于免疫表位数据库(Immune Epitope Database,IEDB)...目的:筛选柯萨奇病毒B3(Coxsackievirus B3,CVB3)中可被细胞毒性T细胞(cytotoxic T lymphocytes,CTL)高效识别的抗原表位,为开发针对CVB3及其他肠道病毒的广谱疫苗提供理论依据。方法:基于免疫表位数据库(Immune Epitope Database,IEDB)及NetMHCpan EL 4.1算法预测CVB3的CTL候选表位;针对HLA-A*02:01限制性表位,通过AlphaFold 2.3.2模拟其与人类白细胞抗原的复合物结构,分析表位保守性及交叉免疫潜力;利用T2细胞结合实验验证候选表位与HLA-A*02:01的亲和力。结果:共鉴定71个潜在CTL表位,筛选出6个HLA-A*02:01高亲和力表位(评分>0.9)。体外实验证实其中5个表位(CTL1~5)可显著诱导HLA-A*02:01表达(P<0.05),而CTL6无结合活性。保守性分析显示,CTL1~6表位在柯萨奇病毒、埃可病毒及肠道病毒中高度保守,提示其交叉保护潜力。结论:鉴定了CVB3的HLA-A*02:01限制性CTL表位,为开发针对CVB3及其他肠道病毒的广谱疫苗提供了关键靶点。展开更多
HLA-A＊02 is the most prevalent and polymorphic major histocompatibility complex （MHC） allele family in humans. Functional differences have been revealed among subtypes, demanding further subtyping of HLA-A＊02 in b...HLA-A＊02 is the most prevalent and polymorphic major histocompatibility complex （MHC） allele family in humans. Functional differences have been revealed among subtypes, demanding further subtyping of HLA-A＊02 in basic and clinical settings. However, the fast growing polymorphisms render traditional primeror probe-based typing methods impractical and result in increasing ambiguities in direct sequence-based typing. In this study, we combined group-specific amplification and mono-allelic sequencing to design and validate a simple scheme for the complete screening and accurate subtyping of all 540 reported HLA-A＊02 alleles. This scheme could be performed in routine labs to facilitate studies with an interest in HLA-A＊02.展开更多
文摘目的:筛选柯萨奇病毒B3(Coxsackievirus B3,CVB3)中可被细胞毒性T细胞(cytotoxic T lymphocytes,CTL)高效识别的抗原表位,为开发针对CVB3及其他肠道病毒的广谱疫苗提供理论依据。方法:基于免疫表位数据库(Immune Epitope Database,IEDB)及NetMHCpan EL 4.1算法预测CVB3的CTL候选表位;针对HLA-A*02:01限制性表位,通过AlphaFold 2.3.2模拟其与人类白细胞抗原的复合物结构,分析表位保守性及交叉免疫潜力;利用T2细胞结合实验验证候选表位与HLA-A*02:01的亲和力。结果:共鉴定71个潜在CTL表位,筛选出6个HLA-A*02:01高亲和力表位(评分>0.9)。体外实验证实其中5个表位(CTL1~5)可显著诱导HLA-A*02:01表达(P<0.05),而CTL6无结合活性。保守性分析显示,CTL1~6表位在柯萨奇病毒、埃可病毒及肠道病毒中高度保守,提示其交叉保护潜力。结论:鉴定了CVB3的HLA-A*02:01限制性CTL表位,为开发针对CVB3及其他肠道病毒的广谱疫苗提供了关键靶点。
基金The authors would like to thank Jueqin Yang for assistance with sample preparation. The authors would also like to thank the Fred Hutchinson Cancer Research Center IHWG Cell and Gene Bank for providing reference genomic DNA samples. This work was supported through grants from the National Natural Science Foundation of China （NSF-30830093） and the National Key Program （973） for Basic Research of China （2009CB522409） to HJ. Supplementary Information accompanies the paper on Cellular ＆ Molecular Immunology website.
文摘HLA-A＊02 is the most prevalent and polymorphic major histocompatibility complex （MHC） allele family in humans. Functional differences have been revealed among subtypes, demanding further subtyping of HLA-A＊02 in basic and clinical settings. However, the fast growing polymorphisms render traditional primeror probe-based typing methods impractical and result in increasing ambiguities in direct sequence-based typing. In this study, we combined group-specific amplification and mono-allelic sequencing to design and validate a simple scheme for the complete screening and accurate subtyping of all 540 reported HLA-A＊02 alleles. This scheme could be performed in routine labs to facilitate studies with an interest in HLA-A＊02.
