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Transcription factor EBF restricts alternative lineage options and promotes B cell fate commitment independently of Pax5

Nature Immunology volume 9, pages 203–215 (2008)Cite this article

Abstract

Alternative lineage restriction and B cell fate commitment require the transcription factor Pax5, but the function of early B cell factor (EBF) in these processes remains mostly unexplored. Here we show that in the absence of EBF, 'expandable' and clonal lymphoid progenitor cells retained considerable myeloid potential. Conversely, ectopic expression of EBF in multipotential progenitor cells directed B cell generation at the expense of myeloid cell fates. EBF induced Pax5 and antagonized expression of genes encoding the transcription factors C/EBPα, PU.1 and Id2. Notably, sustained expression of EBF in Pax5−/− hematopoietic progenitor cells was sufficient to block their myeloid and T lineage potential in vivo. Furthermore, in Pax5−/− pro–B cells, higher EBF expression repressed alternative lineage genes. Thus, EBF can restrict alternative lineage 'choice' and promote commitment to the B cell fate independently of Pax5.

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Figure 1: 'Expandable' and clonal Ebf1−/− lymphoid progenitor cells retain not only T lymphoid but also myeloid developmental potential.
Figure 2: Ebf1−/− lymphoid progenitor cells maintained in vitro in B lymphoid conditions show T cell, NK cell and myeloid developmental potential in vivo.
Figure 3: Higher expression of EBF in MPPs induces B cell development.
Figure 4: EBF promotes B cell fate in MPPs at the expense of myeloid cell fate options.
Figure 7: Constitutive expression of EBF blocks the myeloid lineage differentiation of Pax5−/− pro–B cells.
Figure 5: EBF induces the generation of B cell precursors from myeloid progenitor cells.
Figure 6: EBF inhibits non–B cell fates independently of Pax5.
Figure 8: EBF binds to the Id2 promoter region both in vitro and in vivo.

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Acknowledgements

We thank J.C. Zuniga-Pflucker (University of Toronto) for OP9 and OP9-DL1 cells; M. Busslinger (Institute of Molecular Pathology, Vienna) for Pax5−/− mice and pro–B cells; B. Kee (University of Chicago) for the E2A retroviral constructs; J. Hagman (National Jewish Medical and Research Center) for the epitope-tagged EBF construct; S. Firner (Max Planck Institute of Immunobiology) for doing 'ChIP-on-ChIP' chromatin immunnoprecipitation with EBF; the Immunology Applications Core Facility and the Functional Genomics Facility of the University of Chicago for advice on cell sorting and microarray analysis, respectively; and members of the Singh lab for suggestions and critical comments. Supported by the Irvington Institute (K.L.M.), the Leukemia Lymphoma Society (M.T. and D.A.), the Howard Hughes Medical Institute (H.S.), the National Institute of Allergy and Infectious Diseases and National Institute on Aging of the National Institutes of Health (D.A.) and the German Research Foundation (Deutsche Forschungsgemeinschaft; R.G.).

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Author notes

  1. Kay L Medina

    Present address: Present address: Department of Immunology, The Mayo Clinic, Rochester, Minnesota 55905, USA.,

  2. Jagan M R Pongubala and Daniel L Northrup: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Molecular Genetics and Cell Biology, Howard Hughes Medical Institute, The University of Chicago, Chicago, 60637, Illinois, USA

    Jagan M R Pongubala, David W Lancki, Kay L Medina, Eric Bertolino & Harinder Singh

  2. Department of Pathology and Laboratory of Medicine, University of Pennsylvania School of Medicine, Philadelphia, 19104, Pennsylvania, USA

    Daniel L Northrup, Matthew Thomas & David Allman

  3. Department of Cellular and Molecular Immunology, Max Planck Institute of Immunobiology, Freiburg, 79108, Germany

    Thomas Treiber & Rudolf Grosschedl

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Contributions

J.M.R.P. designed and did experiments, analyzed data and wrote the manuscript; D.L.N. and D.W.L. designed and did experiments and analyzed data; K.L.M., T.T, M.T. and E.B. did experiments; D.A. and R.G. designed and supervised research and contributed to the writing; and H.S. designed and supervised research and wrote the manuscript.

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Correspondence to David Allman or Harinder Singh.

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Pongubala, J., Northrup, D., Lancki, D. et al. Transcription factor EBF restricts alternative lineage options and promotes B cell fate commitment independently of Pax5. Nat Immunol 9, 203–215 (2008). https://doi.org/10.1038/ni1555

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