Isolation and Characterization of Lung Immune Cells

After the lung tissues collected, they were cut into fine pieces and digested with 1× collagenase in DMEM media for 1 h. The cells were passed through a 100-µm filter, and red blood cells were lysed with lysis buffer. The single cells were stained with CD45-AF700, Gr-1-BV605, CD64-PECy7, SiglecF-BV421, CD11b-AF594, CD206-AF488 (BioLegend). The samples were run on Flow Fortessa (BD Biosciences) and data were analyzed by Kaluza Analysis 2.1 (Beckman). The cells were first gated for live single cells followed by gating strategies required for different cell type. For cell sorting, samples were run on Moflo Astrios and cells were collected in DMEM media until further processed. For biodistribution analysis, the different immune cells were gated following a published protocol (55 (link)).