Center for Immunology & Inflammatory Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA.
Harvard Medical School, Boston, Massachusetts, USA.
JCI Insight. 2021 Jul 8;6(13):140028. doi: 10.1172/jci.insight.140028.
Recent advances in high-throughput T cell receptor (TCR) sequencing have allowed for new insights into the human TCR repertoire. However, methods for capturing antigen-specific repertoires remain an area of development. Here, we describe a potentially novel approach that utilizes both a biological and statistical enrichment to define putatively antigen-specific complementarity-determining region 3 (CDR3) repertoires in unselected individuals. The biological enrichment entailed FACS of in vitro antigen-activated memory CD4+ T cells, followed by TCRβ sequencing. The resulting TCRβ sequences were then filtered by selecting those that are statistically enriched when compared with their frequency in the autologous resting T cell compartment. Applying this method to define putatively peanut protein-specific repertoires in 27 peanut-allergic individuals resulted in a library of 7345 unique CDR3β amino acid sequences that had similar characteristics to other validated antigen-specific repertoires in terms of homology and diversity. In-depth analysis of these CDR3βs revealed 36 public sequences that demonstrated high levels of convergent recombination. In a network analysis, the public CDR3βs were shown to be core sequences with more edges than their private counterparts. This method has the potential to be applied to a wide range of T cell-mediated disorders and to yield new biomarkers and biological insights.
高通量 T 细胞受体 (TCR) 测序的最新进展使人们对人类 TCR 库有了新的认识。然而，捕获抗原特异性库的方法仍然是一个有待发展的领域。在这里，我们描述了一种潜在的新方法，该方法利用生物学和统计学富集来定义未选择个体中假定的抗原特异性互补决定区 3 (CDR3) 库。生物学富集包括体外抗原激活的记忆 CD4+ T 细胞的 FACS，然后进行 TCRβ 测序。然后通过选择与自身静息 T 细胞区室中的频率相比统计学上富集的那些来过滤得到的 TCRβ 序列。将该方法应用于定义 27 名花生过敏个体中假定的花生蛋白特异性库，得到了一个包含 7345 个独特 CDR3β 氨基酸序列的文库，这些序列在同源性和多样性方面与其他已验证的抗原特异性库具有相似的特征。对这些 CDR3β 的深入分析揭示了 36 个具有高水平趋同重组的公共序列。在网络分析中，公共 CDR3β 被显示为具有比其私有对应物更多边缘的核心序列。该方法有可能应用于广泛的 T 细胞介导的疾病，并产生新的生物标志物和生物学见解。
