Naïve Regulatory T Cell Subset Is Altered in X-Linked Agammaglobulinemia

Clonosets from 4 cell subsets (CD4+ memory, CD4+ naive, CD8+ memory, CD8+ naive) sorted from blood of XLA and healthy (old and young) donors. Total RNA was isolated using the RNeasy mini kit (Qiagen) according to the manufacturer’s instructions. In all experiments described here, cDNA libraries were obtained using 5’-RACE with unique molecular identifiers (UMI) using Human TCR profiling kit (MiLaboratory LLC). Libraries were sequenced on Illumina NextSeq 500 using 300 cycle reagent kit, paired-end 150 + 150 nt mode. Clonotypes were inferred from reads using MIGEC (UMI threshold = 1) + MIXCR + VDJTOOLS<br>本数据集包含从X连锁无丙种球蛋白血症（XLA）患者及健康老年、青年供者血液中分选得到的4类细胞亚群的克隆型集合（Clonosets），4类亚群分别为CD4+记忆性T细胞、CD4+初始T细胞、CD8+记忆性T细胞与CD8+初始T细胞。总RNA提取严格遵循制造商说明书，采用RNeasy迷你试剂盒（Qiagen）完成。本研究所有实验的cDNA文库均借助带有唯一分子标识符（unique molecular identifiers, UMI）的5'末端快速扩增（5'-RACE）技术，结合人类T细胞受体分型试剂盒（MiLaboratory LLC）构建得到。文库在Illumina NextSeq 500测序平台上，采用300循环试剂盒，以双端150+150 nt的测序模式完成测序。最终通过MIGEC（唯一分子标识符阈值设为1）、MIXCR及VDJTOOLS工具从测序读段中推断得到克隆型。