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首页> 外文期刊>The Journal of Immunology: Official Journal of the American Association of Immunologists >Clustering of class I HLA oligomers with CD8 and TCR:three-dimensional models based on fluorescence resonance energy transfer and crystallographic data
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Clustering of class I HLA oligomers with CD8 and TCR:three-dimensional models based on fluorescence resonance energy transfer and crystallographic data

机译：I类HLA低聚物与CD8和TCR的聚类：基于荧光共振能量转移和晶体学数据的三维模型

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摘要

Fluorescence resonance energy transfer (FRET) data, in accordance with lateral mobility measurements, suggested the existence of class I HLA dimers and oligomers at the surface of live human cells, including the B lymphoblast cell line (JY) used in the present study. Intra- and intermolecular class I HLA epitope distances were measured on JY B cells by FRET using fluorophore-conjugated Ag-binding fragments of mAbs W6/32 and L368 directed against structurally well-characterized heavy and light chain epitopes, respectively. Out-of-plane location of these epitopes relative to the membrane-bound BODIPY-Pc (2-(4,4-difluoro-5-(4..was also deter-mined by FRET. Computer-simulated docking of crystallographic structures of class I HLA and epitope-specific Ag-binding fragments, with experimentally determined interepitope and epitope to cell surface distances as constraints, revealed several sterically allowed and FRET-compatible class I ULA dimeric and tetrameric arrangements. Extension of the tetrameric class I HLA model with interacting TCR and CD8 resulted in a model of a supramolecular cluster that may exist physiologically and serve as a functionally significant unit for a network of CD8-HLA-I complexes providing enhanced signaling efficiency even at low MHC-peptide concentrations at the interface of effector and APCs.
机译：根据横向迁移率测量的荧光共振能量转移（FRET）数据表明，在活人细胞（包括本研究中使用的B淋巴母细胞系）中存在I类HLA二聚体和寡聚体。分子内和分子间I类HLA表位距离是通过FRET在JY B细胞上测量的，使用的是荧光团偶联的mAb W6 / 32和L368的Ag结合片段，分别针对结构良好的重链和轻链表位。这些表位相对于膜结合的BODIPY-Pc（2-（4,4-difluoro-5-（4 ...也由FRET确定）的平面外位置。计算机模拟晶体结构的对接通过实验确定的表位和表位与细胞表面的距离作为约束条件，I类HLA和抗原决定簇特异性的Ag结合片段揭示了几种空间允许且与FRET兼容的I类ULA二聚体和四聚体排列。相互作用的TCR和CD8产生了一个超分子簇模型，该模型可能在生理上存在，并且是CD8-HLA-1配合物网络的功能上重要的单元，即使在效应子和交界处的MHC肽浓度低时也能提供增强的信号传导效率。 APC。

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