Srivastava Ruchi, Khan Arif A, Spencer Doran, Vahed Hawa, Lopes Patricia P, Thai Nhi Thi Uyen, Wang Christine, Pham Thanh T, Huang Jiawei, Scarfone Vanessa M, Nesburn Anthony B, Wechsler Steven L, BenMohamed Lbachir
Laboratory of Cellular and Molecular Immunology, Gavin Herbert Eye Institute, University of California Irvine, School of Medicine, Irvine, CA 92697;
Stem Cell Research Center, University of California Irvine, Irvine, CA 92697;
J Immunol. 2015 Mar 1;194(5):2232-48. doi: 10.4049/jimmunol.1402606. Epub 2015 Jan 23.
The HSV type 1 tegument virion phosphoprotein (VP) 11/12 (VP11/12) is a major Ag targeted by CD8(+) T cells from HSV-seropositive individuals. However, whether and which VP11/12 epitope-specific CD8(+) T cells play a role in the "natural" protection seen in seropositive healthy asymptomatic (ASYMP) individuals (who have never had clinical herpes disease) remain to be determined. In this study, we used multiple prediction computer-assisted algorithms to identify 10 potential HLA-A02:01-restricted CD8(+) T cell epitopes from the 718-aa sequence of VP11/12. Three of 10 epitopes exhibited high-to-moderate binding affinity to HLA-A02:01 molecules. In 10 sequentially studied HLA-A02:01-positive and HSV-1-seropositive ASYMP individuals, the most frequent, robust, and polyfunctional effector CD8(+) T cell responses, as assessed by a combination of tetramer frequency, granzyme B, granzyme K, perforin, CD107(a/b) cytotoxic degranulation, IFN-γ, and multiplex cytokines assays, were predominantly directed against three epitopes: VP11/1266-74, VP11/12220-228, and VP11/12702-710. Interestingly, ASYMP individuals had a significantly higher proportion of CD45RA(low)CCR7(low)CD44(high)CD62L(low)CD27(low)CD28(low)CD8(+) effector memory CD8(+) T cells (TEMs) specific to the three epitopes, compared with symptomatic individuals (with a history of numerous episodes of recurrent ocular herpetic disease). Moreover, immunization of HLA-A02:01 transgenic mice with the three ASYMP CD8(+) TEM cell epitopes induced robust and polyfunctional epitope-specific CD8(+) TEM cells that were associated with a strong protective immunity against ocular herpes infection and disease. Our findings outline phenotypic and functional features of protective HSV-specific CD8(+) T cells that should guide the development of an effective T cell-based herpes vaccine.
单纯疱疹病毒1型（HSV-1）被膜病毒体磷蛋白（VP）11/12是HSV血清阳性个体中CD8⁺T细胞靶向的主要抗原。然而，VP11/12表位特异性CD8⁺T细胞是否以及哪些在血清阳性健康无症状个体（从未患过临床疱疹疾病）中所见的“天然”保护中发挥作用仍有待确定。在本研究中，我们使用多种预测计算机辅助算法从VP11/12的718个氨基酸序列中鉴定出10个潜在的HLA-A02:01限制性CD8⁺T细胞表位。10个表位中的3个对HLA-A02:01分子表现出高至中等的结合亲和力。在10名依次研究的HLA-A02:01阳性且HSV-1血清阳性的无症状个体中，通过四聚体频率、颗粒酶B、颗粒酶K、穿孔素、CD107（a/b）细胞毒性脱颗粒、IFN-γ和多重细胞因子检测组合评估，最常见、最强健和多功能的效应性CD8⁺T细胞反应主要针对3个表位：VP11/12⁶⁶⁻⁷⁴、VP11/12²²⁰⁻²²⁸和VP11/12⁷⁰²⁻⁷¹⁰。有趣的是，与有多次复发性眼部疱疹疾病病史的有症状个体相比，无症状个体中针对这3个表位的CD45RA⁽低⁾CCR7⁽低⁾CD44⁽高⁾CD62L⁽低⁾CD27⁽低⁾CD28⁽低⁾CD8⁺效应性记忆CD8⁺T细胞（TEMs）比例显著更高。此外，用这3个无症状个体的CD8⁺TEM细胞表位免疫HLA-A02:01转基因小鼠可诱导出强健且多功能的表位特异性CD8⁺TEM细胞，这些细胞与针对眼部疱疹感染和疾病的强大保护性免疫相关。我们的研究结果概述了保护性HSV特异性CD8⁺T细胞的表型和功能特征，这些特征应指导有效的基于T细胞的疱疹疫苗的开发。
