PRDM1 shapes germinal center B-cell clonal diversity by gating chromatin accessibility during light-to-dark zone transition

Extensive studies have delineated signaling systems and transcription factors (TFs) that positively regulate germinal center (GC) B cell responses ^1–10 . In the light zone (LZ), T follicular helper (T _fh )-derived CD40L and ICOS along with B cell receptor (BCR) signals license B cell clones to pass through the G1-S checkpoint; the cells then re-enter the dark zone (DZ) for clonal expansion and AID-mediated hypermutation. In contrast, only one signaling-induced negative regulator of the GC response that restricts clonal dominance has been identified, notably Nr4a1 (NUR77) ^11,12 . We posited that PRDM1 (BLIMP1) which promotes exit of GC B cells into the plasma cell (PC) pathway, could also act as a signaling-induced transient feedback gate to restrain the GC B cell response, thereby reducing clonal dominance. In keeping with this hypothesis, low and heterogenous expression of PRDM1 has been reported in GC B cells and its loss has been shown to result in larger GCs ^13,14 .

Using single-cell (sc) transcriptome, V(D)J and chromatin profiling coupled with single nucleotide-resolution accessibility modeling of regulatory DNA sequences, we show that Prdm1 -deficient B cells mount an exaggerated GC reaction characterized by larger clone sizes and enhanced affinity maturation culminating in greater clonal dominance. This phenotype is not attributable merely to impaired exit of GC B cells into the PC pathway. Rather, integrated genomic analyses indicate that PRDM1 constrains expression of genes encoding components of the BCR-signaling cascade. Loss of this feedback augments chromatin engagement of signaling-inducible TFs at ISRE, EICE, NF-κB and POU (Oct) motifs, promoting the G1–S transition during LZ selection and fueling DZ expansion.