Lung airways reprogram circulating memory CD8 T cells

To examine how the transcriptome of circulating memory CD8 T cells isolated from the spleen are reprogrammed in the lung airways, influenza nucleoprotein-specific CD8 T cells from the spleen were FACS sorted and RNA isolated for RNA-seq or the cells were transferred to the lung airways and allowed to rest for 2 days. Following 2 days, donor influenza nucleoprotein-specific CD8 T cells 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. Effector memory (CD44Hi CD62L-) CD8 T cells were isolated from the spleen by cell sorting and transferred intratracheally into naive congenic CD45.1+ mice. 48 hours later, transferred cells were isloated from the airways by bronchoalveolar lavage and purified by sorting on donor (CD45.2+) CD8+ CD44Hi FluNP 366-374 Db tetramer+ populations, and RNA was isolated for downstream analysis.