Abstract
STAT5 and interleukin 7 (IL-7) signaling are thought to control B lymphopoiesis by regulating the expression of key transcription factors and by activating variable (VH) gene segments at the immunoglobulin heavy-chain (Igh) locus. Using conditional mutagenesis to delete the gene encoding the transcription factor STAT5, we demonstrate that the development of pro-B cells was restored by transgenic expression of the prosurvival protein Bcl-2, which compensated for loss of the antiapoptotic protein Mcl-1. Expression of the genes encoding the B cell–specification factor EBF1 and the B cell–commitment protein Pax5 as well as VH gene recombination were normal in STAT5- or IL-7 receptor α-chain (IL-7Rα)-deficient pro-B cells rescued by Bcl-2. STAT5-expressing pro-B cells contained little or no active chromatin at most VH genes. In contrast, rearrangements of the immunoglobulin-κ light-chain locus (Igk) were more abundant in STAT5- or IL-7Rα-deficient pro-B cells. Hence, STAT5 and IL-7 signaling control cell survival and the developmental ordering of immunoglobulin gene rearrangements by suppressing premature Igk recombination in pro-B cells.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$209.00 per year
only $17.42 per issue
Buy this article
Purchase on Springer Link
Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Accession codes
References
von Freeden-Jeffry, U. et al. Lymphopenia in interleukin (IL)-7 gene-deleted mice identifies IL-7 as a nonredundant cytokine. J. Exp. Med. 181, 1519–1526 (1995).
Miller, J.P. et al. The earliest step in B lineage differentiation from common lymphoid progenitors is critically dependent upon interleukin 7. J. Exp. Med. 196, 705–711 (2002).
Kikuchi, K., Lai, A.Y., Hsu, C.-L. & Kondo, M. IL-7 receptor signaling is necessary for stage transition in adult B cell development through up-regulation of EBF. J. Exp. Med. 201, 1197–1203 (2005).
Maraskovsky, E. et al. Bcl-2 can rescue T lymphocyte development in interleukin-7 receptor-deficient mice but not in mutant rag-1−/− mice. Cell 89, 1011–1019 (1997).
Akashi, K., Kondo, M., von Freeden-Jeffry, U., Murray, R. & Weissman, I.L. Bcl-2 rescues T lymphopoiesis in interleukin-7 receptor-deficient mice. Cell 89, 1033–1041 (1997).
Maraskovsky, E., Peschon, J.J., McKenna, H., Teepe, M. & Strasser, A. Overexpression of Bcl-2 does not rescue impaired B lymphopoiesis in IL-7 receptor-deficient mice but can enhance survival of mature B cells. Int. Immunol. 10, 1367–1375 (1998).
Hennighausen, L. & Robinson, G.W. Interpretation of cytokine signaling through the transcription factors STAT5A and STAT5B. Genes Dev. 22, 711–721 (2008).
Teglund, S. et al. Stat5a and Stat5b proteins have essential and nonessential, or redundant, roles in cytokine responses. Cell 93, 841–850 (1998).
Yao, Z. et al. Stat5a/b are essential for normal lymphoid development and differentiation. Proc. Natl. Acad. Sci. USA 103, 1000–1005 (2006).
Hoelbl, A. et al. Clarifying the role of Stat5 in lymphoid development and Abelson-induced transformation. Blood 107, 4898–4906 (2006).
Moriggl, R. et al. Stat5 is required for IL-2-induced cell cycle progression of peripheral T cells. Immunity 10, 249–259 (1999).
Sexl, V. et al. Stat5a/b contribute to interleukin 7-induced B-cell precursor expansion, but abl- and bcr/abl-induced transformation are independent of Stat5. Blood 96, 2277–2283 (2000).
Cui, Y. et al. Inactivation of Stat5 in mouse mammary epithelium during pregnancy reveals distinct functions in cell proliferation, survival, and differentiation. Mol. Cell. Biol. 24, 8037–8047 (2004).
Goetz, C.A., Harmon, I.R., O'Neil, J.J., Burchill, M.A. & Farrar, M.A. STAT5 activation underlies IL7 receptor-dependent B cell development. J. Immunol. 172, 4770–4778 (2004).
Hirokawa, S., Sato, H. Kato, I. & Kudo, A. EBF-regulating Pax5 transcription is enhanced by STAT5 in the early stage of B cells. Eur. J. Immunol. 33, 1824–1829 (2003).
Goetz, C.A. et al. Restricted STAT5 activation dictates appropriate thymic B versus T cell lineage commitment. J. Immunol. 174, 7753–7763 (2005).
Dias, S., Silva, H. Jr., Cumano, A. & Vieira, P. Interleukin-7 is necessary to maintain the B cell potential in common lymphoid progenitors. J. Exp. Med. 201, 971–979 (2005).
Roessler, S. et al. Distinct promoters mediate the regulation of Ebf1 gene expression by IL-7 and Pax5. Mol. Cell. Biol. 27, 579–594 (2007).
Johnston, C.M., Wood, A.L., Bolland, D.J. & Corcoran, A.E. Complete sequence assembly and characterization of the C57BL/6 mouse Ig heavy chain V region. J. Immunol. 176, 4221–4234 (2006).
Chowdhury, D. & Sen, R. Stepwise activation of the immunoglobulin μ heavy chain gene locus. EMBO J. 20, 6394–6403 (2001).
Johnson, K., Angelin-Duclos, C., Park, S. & Calame, K.L. Changes in histone acetylation are associated with differences in accessibility of VH gene segments to V-DJ recombination during B-cell ontogeny and development. Mol. Cell. Biol. 23, 2438–2450 (2003).
Corcoran, A.E., Riddell, A., Krooshoop, D. & Venkitaraman, A.R. Impaired immunoglobulin gene rearrangement in mice lacking the IL-7 receptor. Nature 391, 904–907 (1998).
Bertolino, E. et al. Regulation of interleukin 7-dependent immunoglobulin heavy-chain variable gene rearrangements by transcription factor STAT5. Nat. Immunol. 6, 836–843 (2005).
Igarashi, H., Gregory, S.C., Yokota, T., Sakaguchi, N. & Kincade, P.W. Transcription from the RAG1 locus marks the earliest lymphocyte progenitors in bone marrow. Immunity 17, 117–130 (2002).
McCormack, M.P., Forster, A., Drynan, L., Pannell, R. & Rabbitts, T.H. The LMO2 T-cell oncogene is activated via chromosomal translocations or retroviral insertion during gene therapy but has no mandatory role in normal T-cell development. Mol. Cell. Biol. 23, 9003–9013 (2003).
Kwon, K. et al. Instructive role of the transcription factor E2A in early B lymphopoiesis and germinal center B cell development. Immunity 28, 751–762 (2008).
Yao, Z. et al. Nonredundant roles for Stat5a/b in directly regulating Foxp3. Blood 109, 4368–4375 (2007).
Fleming, H.E. & Paige, C.J. Pre-B cell receptor signaling mediates selective response to IL-7 at the pro-B to pre-B cell transition via an ERK/MAP kinase-dependent pathway. Immunity 15, 521–531 (2001).
Scheeren, F.A. et al. STAT5 regulates the self-renewal capacity and differentiation of human memory B cells and controls Bcl-6 expression. Nat. Immunol. 6, 303–313 (2005).
Opferman, J.T. et al. Development and maintenance of B and T lymphocytes requires antiapoptotic MCL-1. Nature 426, 671–676 (2003).
Schebesta, A. et al. Transcription factor Pax5 activates the chromatin of key genes involved in B cell signaling, adhesion, migration and immune function. Immunity 27, 49–63 (2007).
O'Geen, H., Nicolet, C.M., Blahnik, K., Green, R. & Farnham, P.J. Comparison of sample preparation methods for ChIP-chip assays. Biotechniques 41, 577–580 (2006).
Chakraborty, T. et al. Repeat organization and epigenetic regulation of the DH-Cμ domain of the immunoglobulin heavy-chain gene locus. Mol. Cell 27, 842–850 (2007).
Grawunder, U., Haasner, D., Melchers, F. & Rolink, A. Rearrangement and expression of κ light chain genes can occur without μ heavy chain expression during differentiation of pre-B cells. Int. Immunol. 5, 1609–1618 (1993).
Johnson, K. et al. Regulation of immunoglobulin light-chain recombination by the transcription factor IRF-4 and the attenuation of interleukin-7 signaling. Immunity 28, 335–345 (2008).
Mandal, M. et al. Ras orchestrates exit from the cell cycle and light-chain recombination during early B cell development. Nat. Immunol. 10, 1110–1117 (2009).
Marshall, A.J., Fleming, H.F., Wu, G.E. & Paige, C.J. Modulation of the IL-7 dose-response threshold during pro-B cell differentiation is dependent on pre-B cell receptor expression. J. Immunol. 161, 6038–6045 (1998).
Socolovsky, M., Fallon, A.E., Wang, S., Brugnara, C. & Lodish, H.F. Fetal anemia and apoptosis of red cell progenitors in Stat5a−/−5b−/− mice: a direct role for Stat5 in Bcl-XL induction. Cell 98, 181–191 (1999).
Kieslinger, M. et al. Antiapoptotic activity of Stat5 required during terminal stages of myeloid differentiation. Genes Dev. 14, 232–244 (2000).
Grillot, D.A.M. et al. bcl-x exhibits regulated expression during B cell development and activation and modulates lymphocyte survival in transgenic mice. J. Exp. Med. 183, 381–391 (1996).
Motoyama, N. et al. Massive cell death of immature hematopoietic cells and neurons in bcl-x-deficient mice. Science 267, 1506–1510 (1995).
Decker, T. et al. Stepwise activation of enhancer and promoter regions of the B cell commitment gene Pax5 in early lymphopoiesis. Immunity 30, 508–520 (2009).
Ye, S.-K. et al. The IL-7 receptor controls the accessibility of the TCRγ locus by Stat5 and histone acetylation. Immunity 15, 813–823 (2001).
Rolink, A. et al. A subpopulation of B220+ cells in murine bone marrow does not express CD19 and contains natural killer cell progenitors. J. Exp. Med. 183, 187–194 (1996).
Tudor, K.S., Payne, K.J., Yamashita, Y. & Kincade, P.W. Functional assessment of precursors from murine bone marrow suggests a sequence of early B lineage differentiation events. Immunity 12, 335–345 (2000).
Matthews, A.G. et al. RAG2 PHD finger couples histone H3 lysine 4 trimethylation with V(D)J recombination. Nature 450, 1106–1110 (2007).
Fuxa, M. et al. Pax5 induces V-to-DJ rearrangements and locus contraction of the immunoglobulin heavy-chain gene. Genes Dev. 18, 411–422 (2004).
Schebesta, M., Pfeffer, P.L. & Busslinger, M. Control of pre-BCR signaling by Pax5-dependent activation of the BLNK gene. Immunity 17, 473–485 (2002).
Novobrantseva, T.I. et al. Rearrangement and expression of immunoglobulin light chain genes can precede heavy chain expression during normal B cell development in mice. J. Exp. Med. 189, 75–87 (1999).
Xu, Y., Davidson, L., Alt, F.W. & Baltimore, D. Deletion of the Igκ light chain intronic enhancer/matrix attachment region impairs but does not abolish VκJκ rearrangement. Immunity 4, 377–385 (1996).
Lin, H. & Grosschedl, R. Failure of B-cell differentiation in mice lacking the transcription factor EBF. Nature 376, 263–267 (1995).
Rucker, E.B. III. et al. Bcl-x and Bax regulate mouse primordial germ cell survival and apoptosis during embryogenesis. Mol. Endocrinol. 14, 1038–1052 (2000).
Willis, S.N. et al. Apoptosis initiated when BH3 ligands engage multiple Bcl-2 homologs, not Bax or Bak. Science 315, 856–859 (2007).
Peschon, J.J. et al. Early lymphocyte expansion is severely impaired in interleukin 7 receptor-deficient mice. J. Exp. Med. 180, 1955–1960 (1994).
Shinkai, Y. et al. RAG-2-deficient mice lack mature lymphocytes owing to inability to initiate V(D)J rearrangement. Cell 68, 855–867 (1992).
Ogilvy, S. et al. Constitutive Bcl-2 expression throughout the hematopoietic compartment affects multiple lineages and enhances progenitor cell survival. Proc. Natl. Acad. Sci. USA 96, 14943–14948 (1999).
Yang, Y.W., Model, P. & Heintz, N. Homologous recombination based modification in Escherichia coli and germline transmission in transgenic mice of a bacterial artificial chromosome. Nat. Biotechnol. 15, 859–865 (1997).
Smyth, G.K. & Speed, T. Normalization of cDNA microarray data. Methods 31, 265–273 (2003).
Ren, B. et al. Genome-wide location and function of DNA binding proteins. Science 290, 2306–2309 (2000).
Ji, H. & Wong, W.H. TileMap: create chromosomal map of tiling array hybridizations. Bioinformatics 21, 3629–3636 (2005).
Acknowledgements
We thank L. Hennighausen (National Institutes of Health) for Stat5fl/fl and Bcl2l1fl/fl mice; T. Rabbitts (Leeds Institute of Molecular Medicine) for the Rag1Cre/Cre mouse; J. Adams (Walter and Eliza Hall Institute of Medical Research) for the Vav-Bcl2 mouse; R. Grosschedl (Max-Planck Institute Freiburg) for the Ebf1+/− mouse; L. Hennighausen, R. Moriggl, T. Decker and A. Rolink for discussions; M. Fuxa for advice on immunoglobulin recombination analysis; A. Ebert for help with ChIP analyses; I. Tamir for bioinformatics analysis of ChIP-on-chip data; and G. Stengl for flow cytometry sorting. Supported by Boehringer Ingelheim, the Austrian Science Fund (P16701-BO9), the Austrian GEN-AU initiative (Bundesminsterium für Bildung und Wissenschaft) and the European Union Sixth Framework Programme (funding the Epigenome Network of Excellence and a Marie-Curie fellowship to S. Malin).
Author information
Authors and Affiliations
Contributions
S. Malin did most experiments; S. McManus contributed the ChIP-on-chip analyses; C.C. generated and characterized the Ikzf1Ebf1 transgenic mouse; A.D. did the histological analysis of autoimmune mice; M.N. did the bioinformatics analysis of the Igh locus; P.B. and A.S. provided the Mcl1fl/fl mouse; and M.B. and S. Malin planned the project, designed the experiments and wrote the manuscript.
Corresponding author
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–18 and Supplementary Tables 1–2 (PDF 2886 kb)
Rights and permissions
About this article
Cite this article
Malin, S., McManus, S., Cobaleda, C. et al. Role of STAT5 in controlling cell survival and immunoglobulin gene recombination during pro-B cell development. Nat Immunol 11, 171–179 (2010). https://doi.org/10.1038/ni.1827
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ni.1827
This article is cited by
-
Igh and Igk loci use different folding principles for V gene recombination due to distinct chromosomal architectures of pro-B and pre-B cells
Nature Communications (2023)
-
JAK-STAT signaling pathway-related gene single nucleotide polymorphisms and susceptibility to ankylosing spondylitis in eastern Chinese Han population
Clinical Rheumatology (2023)
-
Activated interleukin-7 receptor signaling drives B-cell acute lymphoblastic leukemia in mice
Leukemia (2022)
-
Interleukin 7 regulates switch transcription in developing B cells
Cellular & Molecular Immunology (2021)
-
Targeting BCL-2 in B-cell malignancies and overcoming therapeutic resistance
Cell Death & Disease (2020)