Monocyte-derived alveolar macrophages induced by influenza infection confer long-term antibacterial protection [RNA-seq]

To investigate transcriptomic differences using RNA-seq in macrophages from a BM origin (CD45.1) and embryonic origin (CD45.2) in influenza-experienced, and naive lungs - at baseline and following stimulation Overall design: In this experiment we sorted alveolar macrophages from naïve, and post-influenza mice using congenic markers to indicate origin. Cells were then stimulated ex vivo for 16 hours with Pam3CSk4 to investigate differences in stimulated cells, but also at steady state (Mock stimulated). This was done in parallel with ATAC-sequencing, to identify changes at both the transcriptional, and chromatin level in the same ells.

本研究旨在通过RNA测序（RNA-seq），探究流感暴露后及未致敏小鼠肺部中，骨髓来源（CD45.1）与胚胎来源（CD45.2）巨噬细胞的转录组差异，检测时点覆盖稳态基线及刺激后状态。
实验设计：本实验通过同源遗传标记（congenic markers）区分细胞来源，从未致敏及流感感染后小鼠体内分选肺泡巨噬细胞；随后将细胞进行离体培养，以Pam3CSK4刺激16小时以探究激活状态下的细胞差异，同时设置Mock刺激组以检测稳态下的细胞特征。本实验同步开展转座酶可及性测序（ATAC-seq），以在同一批细胞中同步解析转录组及染色质层面的变化。