Transcriptome analysis of antigen-specific T follicular helper (Tfh) and non-Tfh cells after Bach2 overexpression

We identified Bach2 as factor to be expressed at low levels in Tfh cells. Induced overexpression of Bach2 in the germinal center reaction results in loss of the Tfh cell population. RNA-seq of sorted antigen-specific Tfh and non-Tfh cells 18 hours after induction of Bach2 overexpression allowed the simultaneous analysis of Bach2 regulated genes and genes differentially expressed in Tfh and non-Tfh cells. Smarta T cell receptor transgenic cells were retrovirally transduced with an inducible vector encoding Bach2 or an empty control vector and transferred into C57BL/6 recipients. Ectopic Bach2 overexpression was induced five days after s.c. immunization with nitrophenol and GP61-80 coupled to mouse serum albumin. After 18 hours, transgenic T cells from pooled inguinal lymph nodes were sorted for a CD44hi CXCR5+ PD-1+ Tfh and CD44hi CXCR5- PD-1- non-Tfh cell phenotype for analysis by RNA-seq.