Direct phenotypic analysis of human MHC class I antigen presentation: Visualization, quanititation, and in situ detection of human viral epitopes using peptide-specific, MHC-restricted human recombinant antibodies
Direct phenotypic analysis of human MHC class I antigen presentation: Visualization, quanititation, and in situ detection of human viral epitopes using peptide-specific, MHC-restricted human recombinant antibodies
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DOI：
10.4049/jimmunol.170.8.4349
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发表时间：
2003-04-15
影响因子：
4.4
通讯作者：
Reiter, Y
中科院分区：
文献类型：
--
作者：
Cohen, CJ;Sarig, O;Reiter, Y
近年来，I类肽-MHC复合物的四聚体阵列的应用使我们能够检测和研究罕见的Ag特异性CD 8（+）T细胞群体。然而，可用的方法不能可视化或确定这些TCR配体在单个细胞上的数量和分布，也不能检测组织中的APC。在这项研究中，我们首次描述了人类I类肽-MHC配体呈递的研究。这些研究通过应用肽特异性、HLA-A2限制性人重组抗体形式的新工具来促进，所述抗体针对源自人T细胞嗜淋巴细胞病毒I型的病毒表位。使用一个大的人抗体噬菌体展示库，我们分离出一个大的面板的重组Fab抗体是特定的肽-MHC I类复合物的肽依赖性，MHC限制性的方式。我们使用这些抗体通过流式细胞术来可视化APC和病毒感染细胞上的特定复合物，以定量APC表面上的特定复合物的数量并原位可视化，所述APC表面上的特定复合物承载由同源病毒Ag的天然存在的活性细胞内加工形成的复合物。这些发现证明了我们将TCR的独特的精细特异性但低固有亲和力转化为对人病毒表位具有TCR样特异性的高亲和力可溶性Ab分子的能力。这些分子可能被证明是至关重要的有用的工具，用于研究MHC I类抗原在健康和疾病中的呈递，以及用于癌症、感染性疾病和自身免疫性疾病的治疗目的。
The advent in recent years of the application of tetrameric arrays of class I peptide-MHC complexes now enables us to detect and study rare populations of Ag-specific CD8(+) T cells. However, aviailable methods cannot visualize or determine the number and distribution of these TCR ligands on individual cells nor detect APCs in tissues. In this study, we describe for the first time studies of human class I peptide-MHC ligand presentation. These studies were facilitated by applying novel tools in the form of peptide-specific, HLA-A2-restricted human recombinant Abs directed toward a viral epitope derived from human T cell lymphotropic virus type I. Using a large human Ab phage display library, we isolated a large panel of recombinant Fab Abs that are specific for a particular peptide-MHC class I complex in a peptide-dependent, MHC-restricted manner. We used these Abs to visualize the specific complex on APCs and virus-infected cells by flow cytometry, to quantify the number of, and visualize in situ, a particular complex on the surface of APCs bearing complexes formed by naturally occurring active intracellular processing of the cognate viral Ag. These findings demonstrate our ability to transform the unique fine specificity, but low intrinsic affinity of TCRs into high affinity soluble Ab molecules endowed with a TCR-like specificity toward human viral epitopes. These molecules may prove to be crucial useful tools for studying MHC class I Ag presentation in health and disease as well as for therapeutic purposes in cancer, infectious diseases, and autoimmune disorders.