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CXCR4 signaling directs Igk recombination and the molecular mechanisms of late B lymphopoiesis


In B lymphopoiesis, activation of the pre-B cell antigen receptor (pre-BCR) is associated with both cell cycle exit and Igk recombination. Yet how the pre-BCR mediates these functions remains unclear. Here, we demonstrate that the pre-BCR initiates a feed-forward amplification loop mediated by the transcription factor interferon regulatory factor 4 and the chemokine receptor C-X-C motif chemokine receptor 4 (CXCR4). CXCR4 ligation by C-X-C motif chemokine ligand 12 activates the mitogen-activated protein kinase extracellular-signal-regulated kinase, which then directs the development of small pre- and immature B cells, including orchestrating cell cycle exit, pre-BCR repression, Igk recombination and BCR expression. In contrast, pre-BCR expression and escape from interleukin-7 have only modest effects on B cell developmental transcriptional and epigenetic programs. These data show a direct and central role for CXCR4 in orchestrating late B cell lymphopoiesis. Furthermore, in the context of previous findings, our data provide a three-receptor system sufficient to recapitulate the essential features of B lymphopoiesis in vitro.

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Fig. 1: Location of proliferating and differentiating pre-B cells in the BM.
Fig. 2: Small pre-B cells are in intimate contact with CXCL12+ stroma.
Fig. 3: Cxcr4 is required for the development of small pre-B cells.
Fig. 4: Pre-B cell differentiation is directly regulated by CXCR4 signaling.
Fig. 5: CXCR4 signaling determines small pre-B cell identity.
Fig. 6: CXCR4 signaling sets the small pre-B cell epigenetic landscape.
Fig. 7: CXCR4 signaling is necessary for Igk recombination.
Fig. 8: ERK signaling downstream of CXCR4 enables pre-B cell differentiation.

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Data availability

The sequences, ATAC-Seq data for WT small pre-B cells, mb1-Cre+Cxcr4fl/fl (Cxcr4 knockout) small pre-B cells, +IL-7, −IL-7 and −IL-7+CXCL12 cultured WT pre-B cells, and −IL-7 and −IL-7+CXCL12 cultured Cxcr4−/− small pre-B cells, as well as all of the corresponding RNA-Seq data, including for +IL-7+CXCL12 and −IL-7+CXCL12+ERK-i cultured pre-B cells, are deposited in the GenBank database (accession number GSE129311).


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We thank M. Olson, R. Ladd and D. Leclerc for cell-sorting services, and the ImmGen Consortium for data assembly. We thank H. Singh (University of Pittsburgh) for providing the PAX5 and FOXO1 ChIP-Seq data, and M. Schlissel (University of Michigan) for providing the -YFP reporter mice. This work is supported by US National Institutes of Health grants AI120715, AI128785 and AI143778 (to M.R.C.), AI120715-02 and AI128785-01A1 (to M.M.), F32AI143120 (to D.E.K.), T32GM007281 (to K.C.M.), UL1TR002003 (to M.M.-C.) and T32HD007009 (to M.K.O.). Part of the bioinformatics analysis was performed by the UIC Research Informatics Core, supported in part by NCATS through grant UL1TR002003.

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Authors and Affiliations



M.M. and M.R.C. designed the experiments. M.M. carried out and analyzed most of the experiments. M.M. and M.M.-C. analyzed the high-throughput sequencing data. M.K.O and D.E.K. assisted with the ATAC-Seq, RNA-Seq and ChIP-Seq experiments, data analyses and flow cytometry. M.V. and K.C.M. performed some sorting and real-time PCR. J.I. and N.K. assisted with the confocal microscopy. I.A. helped to design some of the experiments. F.G. provided the RaDR-GFP mice. M.M. and M.R.C. oversaw the entire project and wrote the final manuscript.

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Correspondence to Malay Mandal or Marcus R. Clark.

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The authors declare no competing interests.

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Peer review information: Laurie Dempsey was the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team.

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Mandal, M., Okoreeh, M.K., Kennedy, D.E. et al. CXCR4 signaling directs Igk recombination and the molecular mechanisms of late B lymphopoiesis. Nat Immunol 20, 1393–1403 (2019).

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