T-cell responses to a specific antigen

The goal of this study is to demonstrate the ability of a novel platform to screen, identify and characterize neoantigen-specific T-cells and their receptors by combining targeted RNA-seq data on subset of T-cell specific genes and T-cell Receptor sequencing Overall design: In this study, we demonstrate the ability to use hybrid-capture based enrichment for a subset of 17 T-cell specific genes on 3' Digital Gene Expression (DGE) RNA-seq libraries. The samples processed were CD8+ T-cells enriched from blood from two independant healthy donors with each pool of CD8+ T-cells being either unstimulated (control) or stimulated with the Cytomegalovirus, Epstein-Barr Virus and Influenza Virus (CEF) control peptide pool overnight to monitor for T-cell activation.

本研究旨在展示一款新型平台的应用能力：该平台可通过结合T细胞特异性基因子集的靶向RNA测序（RNA-seq）数据与T细胞受体（T-cell Receptor, TCR）测序技术，完成新抗原特异性T细胞及其受体的筛选、鉴定与表征。整体实验设计：本研究验证了针对3'端数字基因表达（Digital Gene Expression, DGE）RNA测序文库中的17种T细胞特异性基因子集开展基于杂交捕获的富集的可行性。本次实验处理的样本为从两名独立健康供者血液中富集得到的CD8+ T细胞，每组CD8+ T细胞池均分为未刺激对照组与经巨细胞病毒、EB病毒及流感病毒（CEF）对照肽池过夜刺激的组别，以监测T细胞的激活情况。