Transcriptional profiling of Cd169-lineage lymph node macrophages

Tingible body macrophages (TBMs) exist in lymph node B cell follicles and are responsible for clearing apoptotic B cells. Our data suggests that TBMs are of Cd169-lineage. We isolated cells from Cd169Cre/+.Tdtomatofl/fl reporter mice and transcriptionally profiled them to identify putative TBMs and chacterise their development and function relative to other LN cells of the Cd169 lineage. Cd169Cre/+.Tdtomatofl/fl mice were immunised with hen egg lysozyme (HEL). Lymph nodes were harvested from 2 unimmunised and 2 immunised mice 9 days later. The cells were stained with CITE-seq antibodies and hashtags, and Tomato pos CD11b hi cells were isolated by Fluorescence-activated cell sorting (FACS). As the cells were hashtagged, all samples were combined and split cross two 10X capture reactions

易染体巨噬细胞（tingible body macrophages, TBMs）存在于淋巴结B细胞滤泡中，负责清除凋亡B细胞。我们的研究数据表明，TBMs属于Cd169谱系。我们从Cd169Cre/+·TdTomatofl/fl报告基因小鼠中分离细胞并开展转录组分析，以鉴定潜在的TBMs，并表征其相较于其他Cd169谱系淋巴结细胞的发育过程与功能。我们使用鸡卵溶菌酶（hen egg lysozyme, HEL）免疫Cd169Cre/+·TdTomatofl/fl报告小鼠，分别从2只未免疫小鼠以及免疫后9天的2只小鼠体内采集淋巴结。随后使用CITE-seq抗体与标签对细胞进行染色，并通过荧光激活细胞分选（Fluorescence-activated cell sorting, FACS）分离出TdTomato阳性且CD11b高表达的细胞。由于所有样本均已完成标签标记，我们将所有样本混合后均分至两套10X捕获反应体系中。