Multiplex Identification of Antigen-Specific T Cell Receptors Using a Combination of Immune Assays and Immune Receptor Sequencing.
Multiplex Identification of Antigen-Specific T Cell Receptors Using a Combination of Immune Assays and Immune Receptor Sequencing.
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DOI：
10.1371/journal.pone.0141561
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Faham M
中科院分区：
文献类型：
--
作者：
Klinger M;Pepin F;Wilkins J;Asbury T;Wittkop T;Zheng J;Moorhead M;Faham M
监测抗原特异性T细胞对免疫应答研究、生物标志物和免疫疗法的开发至关重要。我们开发了一种新的多重检测方法，结合了传统的免疫监测技术和免疫受体库测序，能够同时识别对大量抗原特异性的T细胞。我们将30种不同的抗原进行多路混合，并从5个个体中鉴定出427种抗原特异性克隆型，其频率低至百万分之一。鉴定的克隆型通过多种方式进行验证，包括重复性、与已发表的克隆型的一致性以及与ELISPOT的高相关性。应用这项技术，我们已经证明，在不同个体中鉴定出的绝大多数共享抗原特异性克隆型显示出相同的特异性。我们还发现，与对同一抗原特异性的非共享克隆型相比，共享抗原特异性克隆型序列更简单，出现频率更高。总之，该技术能够对数百或数千种抗原的T细胞特异性进行敏感和定量监测，同时以前所未有的分辨率和规模研究T细胞反应。
Monitoring antigen-specific T cells is critical for the study of immune responses and development of biomarkers and immunotherapeutics. We developed a novel multiplex assay that combines conventional immune monitoring techniques and immune receptor repertoire sequencing to enable identification of T cells specific to large numbers of antigens simultaneously. We multiplexed 30 different antigens and identified 427 antigen-specific clonotypes from 5 individuals with frequencies as low as 1 per million T cells. The clonotypes identified were validated several ways including repeatability, concordance with published clonotypes, and high correlation with ELISPOT. Applying this technology we have shown that the vast majority of shared antigen-specific clonotypes identified in different individuals display the same specificity. We also showed that shared antigen-specific clonotypes are simpler sequences and are present at higher frequencies compared to non-shared clonotypes specific to the same antigen. In conclusion this technology enables sensitive and quantitative monitoring of T cells specific for hundreds or thousands of antigens simultaneously allowing the study of T cell responses with an unprecedented resolution and scale.