Detailed and atypical HLA-E peptide binding motifs revealed by a novel peptide exchange binding assay.,European Journal of Immunology

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Detailed and atypical HLA-E peptide binding motifs revealed by a novel peptide exchange binding assay.
European Journal of Immunology ( IF 4.5 ) Pub Date : 2020-07-27 , DOI: 10.1002/eji.202048719
Lucy C Walters ₁ , Andrew J McMichael ₁ , Geraldine M Gillespie ₁

Affiliation

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, several Mycobacterial peptides 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.

中文翻译：

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。

最近，在使用重组恒河巨细胞病毒 (RhCMV 68-1) 载体的 SIV 疫苗研究中，发现了与 HLA-E 恒河猴同源物 Mamu-E 结合的多种 SIV 和 HIV 表位，从而实现了针对 SIV 攻击的前所未有的保护。此外，通过算法和从受感染细胞洗脱后鉴定的几种分枝杆菌肽，通过人类 HLA-E 呈递给 CD8 ⁺ T 细胞。然而，目前还缺乏通过可靠、高通量的体外测定对 HLA-E 肽相对结合强度进行比较和全面的分析。为了解决这个问题，我们开发并优化了一种新型、高灵敏度的基于肽交换 ELISA 的检测方法，可以相对定量与 HLA-E 的肽结合。使用这种方法，我们筛选了多种肽，包括来自 HIV、SIV 和 Mtb 的肽组，预计可结合 HLA-E。我们的结果表明，虽然 HLA-E 优先容纳规范的 MHC I 类前导肽，但许多非规范的、序列多样的、病原体衍生的肽也结合 HLA-E，尽管通常具有较低的相对结合强度。此外，我们的筛选表明，大多数测试的肽，包括一些关键的 Mtb 和 SIV 表位，已被证明可以引发强烈的 Mamu-E 限制性 T 细胞反应，要么与 HLA-E 结合极弱，要么发出与 HLA-E 无法区分的信号。阴性、无肽对照。

更新日期：2020-07-27

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