Detailed and atypical HLA-E peptide binding motifs revealed by a novel peptide exchange binding assay.
Detailed and atypical HLA-E peptide binding motifs revealed by a novel peptide exchange binding assay.
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新型肽交换结合测定揭示了详细且非典型的 HLA-E 肽结合基序。
DOI：
10.1002/eji.202048719
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发表时间：
2020
影响因子：
5.4
通讯作者：
Walters LC
中科院分区：
文献类型：
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作者：
Walters LC
最近，在使用重组猕猴巨细胞病毒(RhCMV 68-1)载体的SIV疫苗研究中，发现了与恒河猴HLAE同源物Mamu-E结合的多种SIV和HIV表位，对SIV攻击获得了前所未有的保护。此外，几种通过算法和从感染细胞洗脱后鉴定的分枝杆菌多肽被人类的HLA-E呈递给CD8+T细胞。然而，通过一种可靠的、高通量的体外试验来比较和全面地分析相对的人类白细胞抗原-E多肽结合强度目前还缺乏。为了解决这个问题，我们开发和优化了一种新的、高灵敏的基于肽交换的ELISA法，它可以相对定量地与HLA-E结合。使用这种方法，我们筛选了多个肽，包括来自HIV、SIV和Mtb的多肽，预测可以与HLA-E结合。我们的结果表明，尽管HLA-E优先容纳规范的MHC I类先导肽，但许多非规范的、序列多样的病原体衍生的肽也与HLA-E结合，尽管通常相对结合强度较低。此外，我们的筛查显示，大多数被测试的多肽，包括一些关键的Mtb和SIV表位，已经被证明能激发强烈的Mamu-E限制性T细胞反应，要么与HLA-E结合得非常弱，要么给出与阴性的无肽对照难以区分的信号。
Diverse SIV and HIV epitopes that bind the rhesus homolog of HLA‐E, Mamu‐E, have recently been identified in SIVvaccine studies using a recombinant Rhesus cytomegalovirus (RhCMV 68‐1) vector, where unprecedented protection against SIV challenge was achieved. Additionally, severalMycobacterialpeptides identified both algorithmically and following elution from infected cells, are presented to CD8+T cells by HLA‐E in humans. Yet, a comparative and comprehensive analysis of relative HLA‐E peptide binding strength via a reliable, high throughput in vitro assay is currently lacking. To address this, we developed and optimized a novel, highly sensitive peptide exchange ELISA‐based assay that relatively quantitates peptide binding to HLA‐E. Using this approach, we screened multiple peptides, including peptide panels derived from HIV, SIV, and Mtb predicted to bind HLA‐E. Our results indicate that although HLA‐E preferentially accommodates canonical MHC class I leader peptides, many non‐canonical, sequence diverse, pathogen‐derived peptides also bind HLA‐E, albeit generally with lower relative binding strength. Additionally, our screens demonstrate that the majority of peptides tested, including some key Mtb and SIV epitopes that have been shown to elicit strong Mamu‐E‐restricted T cell responses, either bind HLA‐E extremely weakly or give signals that are indistinguishable from the negative, peptide‐free controls.