IRF4 regulates the proliferative response during B cell differentiation in vivo

Abstract

Abstract Cell division is an essential component of B cell differentiation to a plasma cell (PC). Division coupled changes in the expression of transcription factors that coordinate the PC program are known; however, little is known regarding the timing/extent of reprogramming by these factors as the cells are dividing in vivo. Furthermore, how/when these factors coordinate cell division during differentiation has remained unexplored. To address this, we assessed the cell division kinetics of wild-type (WT) B cells responding to LPS in vivo. Interestingly, we found that WT B cells must undergo 8 divisions before differentiating into PCs. Computational modeling of division rates defined a proliferative burst between 48 and 60 hours after LPS injection that is characterized by a rapid increase in the rate of cell division. In contrast, Interferon Regulatory Factor 4-deficient (IRF4−/−) B cells divided but stalled at divisions 2–6, failing to undergo the proliferative burst. To assess the timing/scope of IRF4-dependent reprogramming, WT and IRF4−/− responding B cells at divisions 0, 1, and 3–6 were sorted for ATAC- and RNA-seq. RNA-seq data revealed hundreds of differentially expressed genes (DEGs) when IRF4 was deleted, a subset of which included Myc target genes. ATAC-seq data also exposed hundreds of differentially accessible regions (DARs), the majority of which contained a known IRF4 binding motif and mapped to a corresponding DEG. Interestingly, IRF4−/− B cells progressively became more divergent as they divided when compared to WT B cells in the same division. Together, these data create a road map defining the role of IRF4 throughout B cell differentiation and reveal a critical role for IRF4 in maintaining the proliferative response.