Dissection of memory response B cell populations by single cell RNA-seq

SWHEL B cells were adoptively transferred and recipient mice were immunised with hen egg lysozyme (HEL) to establish anti-HEL memory B cells. Immune mice were then re-immunised with HEL and single donor-derived antigen-specific B cells sorted for RNA sequencing on day 5 of the secondary antibody response. Overall design: Single-cell resolution transcriptional profiles of 144 memory response (Day 5 post-recall) SWHEL-Tomato B cells from three mice. Single-cell samples were isolated by FACS and processed in 9 batches. Samples were split into two pools of multiplexed samples. Each pool was run on a single Illumina HiSeq 2500 lane, averaging 3 million reads per cell.

将SWHEL B细胞过继转移至受体小鼠，并用鸡溶菌酶（hen egg lysozyme, HEL）免疫受体小鼠，以构建抗HEL记忆B细胞模型。随后对已免疫的小鼠再次施以HEL免疫，并在二次抗体应答的第5天，分选供体来源的抗原特异性单个B细胞用于RNA测序。
实验整体设计：本数据集包含来自3只小鼠的144个记忆应答（召回免疫后第5天）SWHEL-Tomato B细胞的单细胞分辨率转录组图谱。单细胞样本通过荧光激活细胞分选术（FACS）分离，并分为9个批次进行处理。样本被划分为两个多重测序样本池，每个样本池在一条Illumina HiSeq 2500测序通道上完成测序，每个细胞平均获取300万条测序读段。