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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Epitope discovery in West Nile virus infection: Identification and immune recognition of viral epitopes
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Epitope discovery in West Nile virus infection: Identification and immune recognition of viral epitopes

机译：西尼罗河病毒感染中的表位发现：病毒表位的鉴定和免疫识别

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摘要

Cytotoxic T lymphocytes (CTL) play an important role in the control and elimination of infection by West Nile virus (WNV), yet the class I human leukocyte antigen (HLA)-presented peptide epitopes that enable CTL recognition of WNV-infected cells remain uncharacter-ized. The goals of this work were first to discover the peptide epitopes that distinguish the class I HLA of WNV-infected cells and then to test the T cell reactivity of newly discovered WNV epitopes. To discover WNV-immune epitopes, class I HLA was harvested from WNV (NY99 strain)-infected and uninfected HeLa cells. Then peptide epitopes were eluted from affinity-purified HLA, and peptide epitopes from infected and uninfected cells were comparatively mapped by mass spectroscopy. Six virus-derived peptides from five different viral proteins (E, NS2b, NS3, NS4b, and NS5) were discovered as unique to HLA-A~*0201 of infected cells, demonstrating that the peptides sampled by class I HLA are distributed widely throughout the WNV proteome. When tested with CTL from infected individuals, one dominant WNV target was apparent, two epitopes were subdominant, and three demonstrated little CTL reactivity. Finally, a sequence comparison of these epitopes with the hundreds of viral isolates shows that HLA-A~*0201 presents epitopes derived from conserved regions of the virus. Detection and recovery from WNV infection are therefore functions of the ability of class I HLA molecules to reveal conserved WNV epitopes to an intact cellular immune system that subsequently recognizes infected cells.
机译：细胞毒性T淋巴细胞（CTL）在控制和消除西尼罗河病毒（WNV）的感染中起着重要作用，但I类人类白细胞抗原（HLA）呈递的肽表位仍使CTL识别感染了WNV的细胞仍然没有特征-这项工作的目标是首先发现区分WNV感染细胞的I类HLA的肽表位，然后测试新发现的WNV表位的T细胞反应性。为了发现WNV免疫表位，从WNV（NY99株）感染和未感染的HeLa细胞中收获I类HLA。然后从亲和纯化的HLA中洗脱出肽表位，并通过质谱法比较绘制了感染和未感染细胞的肽表位。发现来自五种不同病毒蛋白（E，NS2b，NS3，NS4b和NS5）的六种病毒衍生肽是感染细胞的HLA-A〜* 0201所特有的，这表明I类HLA所采样的肽广泛分布WNV蛋白质组。当用来自感染个体的CTL进行测试时，一个明显的WNV靶标很明显，两个表位占优势，三个CTL反应性很低。最后，这些表位与数百种病毒分离株的序列比较表明，HLA-A〜* 0201呈现出源自病毒保守区的表位。因此，从WNV感染中进行检测和恢复是I类HLA分子向完整的细胞免疫系统揭示保守WNV表位的能力的功能，该细胞免疫系统随后识别受感染的细胞。

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