Gene expression analysis of macrophages exposed to LPS and LPS-coated beads

The aim of this study was to measure the response of alveolar macrophages (AM) and bone marrow-derived macrophages (BMDM) to LPS, LPS-coated beads, and PBS-coated beads Overall design: Mice were exposed to LPS-coated beads or PBS-coated beads using a nebulizer. Alveolar macrophages were isolated 4 hours after exposure by broncholaveolar lavage and by fluorescence activated cell sorting using the following surface markers (CD45+, Zombie Violet-, CD3-/CD19-, Siglec F+, CD64+, FITC-bead+ or FITC-bead-). Bone marrow-derived macrophages were collected 4 hours after exposure to LPS-coated beads, PBS-coated beads, or soluble LPS by fluorescence activated cell sorting using the following surface markers (CD45+, Zombie Violet-, CD3-/CD19-, F4/80+, FITC-bead+ or FITC-bead-). Samples were analyzed by RNA-sequencing.

本研究旨在检测肺泡巨噬细胞（alveolar macrophages, AM）与骨髓来源巨噬细胞（bone marrow-derived macrophages, BMDM）对脂多糖（LPS）、脂多糖包被微球以及磷酸盐缓冲液（PBS）包被微球的应答反应。实验设计概况：通过雾化器使小鼠暴露于脂多糖包被微球或磷酸盐缓冲液包被微球环境中。暴露4小时后，经支气管肺泡灌洗分离肺泡巨噬细胞，并通过以下表面标志物采用荧光激活细胞分选进行纯化：CD45+、Zombie Violet-、CD3-/CD19-、Siglec F+、CD64+、FITC标记微球阳性或FITC标记微球阴性。骨髓来源巨噬细胞则在暴露于脂多糖包被微球、磷酸盐缓冲液包被微球或可溶性脂多糖4小时后，通过以下表面标志物采用荧光激活细胞分选进行收集：CD45+、Zombie Violet-、CD3-/CD19-、F4/80+、FITC标记微球阳性或FITC标记微球阴性。所有样本均通过RNA测序（RNA-sequencing）完成分析。