KaedeGreen and Red CD8 T cells from vaccinia-GP33 infected skin

In order to determine transcriptional differences between cells in the draining lymph node and those recently emigrating from the skin, we performed single cell RNA sequencing on KaedeGreen and KaedeRed CD8 T cells 12 days post infection with vaccinia-GP33. CD8+ cells were MACS enriched. Cells were stained with two tetramers (H2-Kb-B8R20-27 and H2-Db-GP33-41) conjugated to APC. Cells were then incubated with TotalSeq anti-APC (Biolegend, 408009) Overall design: Kaede male and female mice were infected with 5x10^6 PFU of VV-GP33 on both ears via scarification. 11 days later ears were photoconverted with 405nm light. 24 hours after photoconversion, CD8 T cells were isolated from draining lymph nodes. Cells were sorted on CD8a+, CD44+, CD45+, Intravascular CD45-, Live (DAPI-).