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Foxp1 is an essential transcriptional regulator of B cell development

Nature Immunology volume 7, pages 819–826 (2006)Cite this article

Abstract

Forkhead transcription factors are key participants in development and immune regulation. Here we demonstrate that absence of the gene encoding the forkhead transcription factor Foxp1 resulted in a profound defect in early B cell development. Foxp1 deficiency was associated with decreased expression of all B lineage genes in B220+ fetal liver cells as well as with a block in the transition from pro–B cell to pre–B cell involving diminished expression of recombination-activating genes 1 and 2. Foxp1 bound to the Erag enhancer and was involved in controlling variable-(diversity)-joining recombination of the gene encoding immunoglobulin heavy chain in a B cell lineage–specific way. Our results identify Foxp1 as an essential participant in the transcriptional regulatory network of B lymphopoiesis.

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Figure 1: The generation of Foxp1−/− mice.
Figure 2: B cell development is impaired in the absence of Foxp1.
Figure 3: Impaired μ-chain generation in Foxp1−/− pro–B cells ex vivo.
Figure 4: Expression of B cell lineage genes in B220+ Foxp1−/− fetal liver cells and pro–B cells cultured in vitro.
Figure 5: Foxp1 binds directly to consensus forkhead sites in the Erag enhancer, as shown by chromatin immunoprecipitation and EMSA.
Figure 6: V(D)J rearrangement of Igh genes is impaired in Foxp1−/− pro–B cells cultured in vitro.

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Acknowledgements

We thank K. Rajewsky, S. Casola, K. Otipoby, C. Xiao, G. Galler, K. Mark Ansel and H. Liu for reagents and discussions; S. Monticelli for critical reading of the manuscript; B. Tanasa for help with the bioinformatic analyses; L. Smith for preparing genomic DNA for genotyping; C. Das and C. Schmidt for technical help; and N. Barteneva and K. Ketman for flow cytometry. Supported by the National Institutes of Health (T32 to H.H.; CA42471, AI48213 and AI44432 to A.R.; and HL071160, CA92318 and CA31534 to P.W.T.) and the Mary Betzner Morrow Centennial endowment in Molecular Genetics (P.W.T.).

Author information

Author notes

  1. Bin Wang

    Present address: Department of Physiology, University of Texas Health Science Center at San Antonio, San Antonio, Texas, 78229, USA

  2. Julie Nardone

    Present address: Cell Signaling Technology, Danvers, Massachusetts, 01923, USA

Authors and Affiliations

  1. Department of Pathology, Harvard Medical School, CBR Institute for Biomedical Research, Boston, 02115, Massachusetts, USA

    Hui Hu, Madhuri Borde, Julie Nardone & Anjana Rao

  2. Department of Molecular Genetics and The Institute for Cellular and Molecular Biology, The University of Texas at Austin, Austin, 78712, Texas, USA

    Bin Wang, Shan Maika, Laura Allred & Philip W Tucker

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Contributions

H.H. and A.R. initiated the collaboration with P.W.T. to analyze the immune phenotype of Foxp1−/− mice; B.W. and S.M. generated the Foxp1−/− mice; H.H. was responsible for all analyses of B cell development in fetal liver chimeras and in vitro pro–B cell cultures, including analysis of Foxp1 expression and chromatin immunoprecipitation assays; L.A. did EMSA and footprinting experiments under the supervision of P.W.T.; M.B. helped with mouse breeding and fetal liver collection; J.N. helped with bioinformatic analysis; and A.R. and P.W.T. provided overall supervision.

Corresponding authors

Correspondence to Hui Hu or Anjana Rao.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Fig. 1

Gene targeting strategy, expression pattern of Foxp1 at different B cell developmental stages and the kinetics of peripheral B cell recovery in reconstituted RAG2-deficient recipient mice. (PDF 688 kb)

Supplementary Fig. 2

No difference in total peripheral B cell numbers between Foxp1+/+ and Foxp1+/− mice. (PDF 378 kb)

Supplementary Fig. 3

Foxp1−/− thymocyte development. (PDF 415 kb)

Supplementary Fig. 4

Expression of IL-7R on Foxp1+/− or Foxp1−/− pro-B cells and Foxp1 ChIP in thymocytes. (PDF 354 kb)

Supplementary Fig. 5

Identification of Foxp1 binding sites within KpnI-PvuII region by DNase I footprinting and EMSA. (PDF 1430 kb)

Supplementary Fig. 6

Donor cell recovery in reconstituted RAG2-deficient recipients. (PDF 410 kb)

Supplementary Table 1

Sequences of primers for Foxp1 expression and B lineage gene expression. (PDF 56 kb)

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Hu, H., Wang, B., Borde, M. et al. Foxp1 is an essential transcriptional regulator of B cell development. Nat Immunol 7, 819–826 (2006). https://doi.org/10.1038/ni1358

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