Gene expression profile (targeted mouse immune response panel) and V(D)J sequences at single cell level of day 10 post-IAV infected splenic T cells in WT mice and LCKFREE mice

The LckC20.23A (LCKFREE) mutation in mice dissociates LCK from the CD4 and CD8 coreceptors. We compared the transcriptional profile of conventional splenic CD8 and CD4 T cell subsets at day 10 post-IAV infection of LCKFREE mice to WT mice to assess the functional impact of dissociating LCK from coreceptors.. 3xC57Bl/6 and 3x LckC20.23A aged 8 weeks old were infected intranasally with 10E4 pfu HKx31 IAV infection and their spleens harvested at day 10 post-infection. Mouse splenocytes were stained and sorted using BD® FACSAria™ Fusion (BD Biosciences) into CD8 TN, TVM, and effector/memory T (TE/EM) cells and CD4 TN and TE/EM cells based on expression of CD44, CD49d, and CD62L. Sorted populations from the same individual were pooled and sample-tagged using the BD® Single-Cell Multiplexing Kit. Single-cell capture and cDNA synthesis were performed using the BD Rhapsody™ Single-Cell Analysis System. Targeted scRNA-seq analysis was performed using the BD Rhapsody™ Immune Response Targeted Panel (mouse) containing primers of 397 immune-related genes. A paired TCRαβ sequence library was prepared using the BD Rhapsody™ Mouse TCR/BCR Amplification Kit.