Transcriptome profiling of influenza-specific memory CD8 T cells in the lung airways, lung parenchyma, and spleen

To examine differences in the transcriptome of resident memory CD8 T cells in the lung, influenza nucleoprotein-specific CD8 T cells from the airways , lung parenchyma, and spleen were FACS sorted and RNA isolated for RNA-seq. C57Bl/6 mice were infected with influenza x31 and rested for 35 days to establish T cell memory. Five minutes prior to euthanasia mice were injected with anti-CD45.2-PE to discriminate between tissue resident and circulating cells, and cells were isolated from the airways by bronchoalveolar lavage, from the lung parechyma by enzymatic digestion, and from the spleen by mechanical disruption. Cells were sorted on CD8+ CD44Hi FluNP 366-374 Db tetramer+ populations, and cells from the airways and parenchyma were further sorted on intravital label negative (i.e. tissue resident) cells.