Empty peptide-receptive MHC class I molecules for efficient detection of antigen-specific T cells
Empty peptide-receptive MHC class I molecules for efficient detection of antigen-specific T cells
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DOI：
10.1126/sciimmunol.aau9039
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发表时间：
2019-07-01
影响因子：
24.8
通讯作者：
Hadrup, Sine Reker
中科院分区：
文献类型：
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作者：
Saini, Sunil Kumar;Tamhane, Tripti;Hadrup, Sine Reker
MHC I类分子的肽依赖性稳定性对它们在基于肽-MHC多聚体的方法中用于全面分析T细胞免疫提出了实质性挑战。为了克服这一挑战，我们证明了使用功能空的MHC I类分子通过二硫键稳定连接α 1和α（2）螺旋接近F口袋。负载肽的二硫键稳定的HLA-A*02：01显示与野生型HLA-A*02：01的完全结构重叠。使用二硫化物稳定的HLA-A*02：01、HLA-A*24：02和H-2K（B）制备的肽-MHC多聚体可用于鉴定抗原特异性T细胞，并且与用野生型MHC I类分子制备的多聚体相比，它们为抗原特异性T细胞检测提供更好的染色指数。二硫键稳定的MHC I类分子可以负载多聚化形式的肽，而不影响其染色T细胞的能力。我们证明了通过单步肽添加将空泡四聚体转化为抗原特异性四聚体的价值，其用于鉴定对人黑色素瘤中突变衍生的新抗原和其他癌症相关抗原特异性的T细胞。
The peptide-dependent stability of MHC class I molecules poses a substantial challenge for their use in peptide-MHC multimer-based approaches to comprehensively analyze T cell immunity. To overcome this challenge, we demonstrate the use of functionally empty MHC class I molecules stabilized by a disulfide bond to link the alpha 1 and alpha(2) helices close to the F pocket. Peptide-loaded disulfide-stabilized HLA-A*02:01 shows complete structural overlap with wild-type HLA-A*02:01. Peptide-MHC multimers prepared using disulfide-stabilized HLA-A*02:01, HLA-A*24:02, and H-2K(b) can be used to identify antigen-specific T cells, and they provide a better staining index for antigen-specific T cell detection compared with multimers prepared with wild-type MHC class I molecules. Disulfide-stabilized MHC class I molecules can be loaded with peptide in the multimerized form without affecting their capacity to stain T cells. We demonstrate the value of empty-loadable tetramers that are converted to antigen-specific tetramers by a single-step peptide addition through their use to identify T cells specific for mutation-derived neoantigens and other cancer-associated antigens in human melanoma.