Developmental dynamics and genomic programming of GC-derived precursors that give rise to long lived plasma cells

Abstract

Abstract Plasma cell (PC) precursors are generated during a germinal center (GC) response and migrate to the bone marrow (BM) where they undergo terminal differentiation into short-lived (SL) or long-lived (LL) antibody secreting cells. The developmental dynamics and genomic states of PC precursors remain to be elucidated1. We utilize a novel experimental system, involving CD138+ splenocytes isolated from NP-KLH immunized C57Bl/6J mice (21–42 dpi) and their adoptive transfer into B cell deficient mMT mice, to analyze developmental dynamics of antigen-specific PC precursors and their progeny at single-cell resolution. We demonstrate that SLPC precursors are generated earlier (d21), whilst LLPC precursors are generated later (d35) during a GC response. scRNA-seq analyses reveals increased frequency of a novel cluster of transitional cells expressing both B-lineage and plasma cell-specific genes. Coupled analyses using BCR-seq shows that NP-specific transitional cells express cell cycle genes and undergo greater clonal expansion on d35 to give rise to proliferating and quiescent PCs. Antigen-specific clonal tracking in the spleen and BM compartments suggests that the transitional cells emanating from the GC, differentiate into proliferating PCs before migrating to the BM. CITE-seq analyses and reconstitution experiments demonstrate that proliferating PCs are contained within the B220− CD138+ CD44+ CD11a+ subset and are enriched for BMPC precursors. These results reveal novel GC-derived transitional cells that generate precursors of both SLPCs and LLPCs with differing developmental and proliferation dynamics in a humoral immune response.