Comprehensive Evaluation of the Expressed CD8+T Cell Epitope Space Using High-Throughput Epitope Mapping
Comprehensive Evaluation of the Expressed CD8+T Cell Epitope Space Using High-Throughput Epitope Mapping
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DOI：
10.3389/fimmu.2019.00655
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发表时间：
2019-04-26
影响因子：
7.3
通讯作者：
Reche, Pedro A.
中科院分区：
文献类型：
--
作者：
Lehmann, Paul, V;Suwansaard, Maneewan;Reche, Pedro A.
T细胞免疫传统上通过功能回忆测定来评估，其检测T细胞抗原遭遇的结果，或通过选择性结合肽特异性T细胞受体的荧光标记的多聚体来评估。使用任一种方法，如果复杂抗原系统（如病毒）的错误抗原或肽用于免疫监测，则将获得假阴性数据，或者抗原特异性T细胞区室的大小将被大大低估。在这项工作中，我们展示了如何选择“正确的”抗原或抗原肽是成功的T细胞免疫监测对人巨细胞病毒（HCMV）的关键。具体而言，我们证明，个别HCMV抗原，沿着与以前报道的表位，经常未能检测到CD8+ T细胞免疫试验对象。通过系统评估T细胞对衍生自HCMVpp65蛋白的单个九聚体肽的反应性，我们的数据清楚地建立了（i）需要对由感兴趣的抗原的基因组编码的所有潜在表位进行系统测试以可靠地检测CD8+ T细胞免疫，和（ii）全基因组，通过基于ELISPOT的高通量“蛮力”表位作图，肽的大规模系统测试已经变得可行。
T cell immunity is traditionally assessed through functional recall assays, which detect the consequences of the T cells' antigen encounter, or via fluorescently labeled multimers that selectively bind peptide-specific T cell receptors. Using either approach, if the wrong antigen or peptide of a complex antigenic system, such as a virus, is used for immune monitoring, either false negative data will be obtained, or the magnitude of the antigen-specific T cell compartment will go largely underestimated. In this work, we show how selection of the "right" antigen or antigenic peptides is critical for successful T cell immune monitoring against human cytomegalovirus (HCMV). Specifically, we demonstrate that individual HCMV antigens, along with previously reported epitopes, frequently failed to detect CD8+ T cell immunity in test subjects. Through systematic assessment of T cell reactivity against individual nonamer peptides derived from the HCMVpp65 protein, our data clearly establish that (i) systematic testing against all potential epitopes encoded by the genome of the antigen of interest is required to reliably detect CD8+ T cell immunity, and (ii) genome-wide, large scale systematic testing of peptides has become feasible through high- throughput ELISPOT-based "brute force" epitope mapping.