Evolution of antigen-specific follicular helper T cell transcriptional progams across effector function and through to memory

We utilized single cell-indexed custom RNA-sequencing to interogate the transcriptomes of TFH cells at Day 7, Day 14, and Day 28 following NP-PCC immunization Overall design: T cells were isolated from draining lymph nodes of B10.Br mice following base of tail immunization with NP-PCC in MPL at Day 7, Day 14, and D28. Phenotypically defined T cell subsets sorted as: naïve CD4 T cells (CD4+, CD8- CD62L+, CD44-), Effector T helper cells (CD4+, CD8- CD44+ CD62L-, CXCR5-), T follicular cells (CD4+, CD8-, CD44+. CD62L-, CXCR5+). Cells were sorted into 384 well plates for library preparation. cDNA was amplied using a nested set of ~500 gene specific targeted primers.