Abstract
The stepwise commitment from hematopoietic stem cells in the bone marrow to T lymphocyte–restricted progenitors in the thymus represents a paradigm for understanding the requirement for distinct extrinsic cues during different stages of lineage restriction from multipotent to lineage-restricted progenitors. However, the commitment stage at which progenitors migrate from the bone marrow to the thymus remains unclear. Here we provide functional and molecular evidence at the single-cell level that the earliest progenitors in the neonatal thymus had combined granulocyte-monocyte, T lymphocyte and B lymphocyte lineage potential but not megakaryocyte-erythroid lineage potential. These potentials were identical to those of candidate thymus-seeding progenitors in the bone marrow, which were closely related at the molecular level. Our findings establish the distinct lineage-restriction stage at which the T cell lineage–commitment process transits from the bone marrow to the remote thymus.
Similar content being viewed by others
Accession codes
References
Reya, T., Morrison, S.J., Clarke, M.F. & Weissman, I.L. Stem cells, cancer, and cancer stem cells. Nature 414, 105–111 (2001).
Katsura, Y. Redefinition of lymphoid progenitors. Nat. Rev. Immunol. 2, 127–132 (2002).
Donskoy, E. & Goldschneider, I. Thymocytopoiesis is maintained by blood-borne precursors throughout postnatal life. A study in parabiotic mice. J. Immunol. 148, 1604–1612 (1992).
Scollay, R., Smith, J. & Stauffer, V. Dynamics of early T cells: prothymocyte migration and proliferation in the adult mouse thymus. Immunol. Rev. 91, 129–157 (1986).
Allman, D. et al. Thymopoiesis independent of common lymphoid progenitors. Nat. Immunol. 4, 168–174 (2003).
Benz, C. & Bleul, C.C. A multipotent precursor in the thymus maps to the branching point of the T versus B lineage decision. J. Exp. Med. 202, 21–31 (2005).
Bhandoola, A., von Boehmer, H., Petrie, H.T. & Zuniga-Pflucker, J.C. Commitment and developmental potential of extrathymic and intrathymic T cell precursors: plenty to choose from. Immunity 26, 678–689 (2007).
Wada, H. et al. Adult T-cell progenitors retain myeloid potential. Nature 452, 768–772 (2008).
Bell, J.J. & Bhandoola, A. The earliest thymic progenitors for T cells possess myeloid lineage potential. Nature 452, 764–767 (2008).
Desanti, G.E. et al. Clonal analysis reveals uniformity in the molecular profile and lineage potential of CCR9+ and CCR9− thymus-settling progenitors. J. Immunol. 186, 5227–5235 10.4049/jimmunol.1002686 (2011).
Masuda, K. et al. Thymic anlage is colonized by progenitors restricted to T, NK, and dendritic cell lineages. J. Immunol. 174, 2525–2532 (2005).
Kondo, M., Weissman, I.L. & Akashi, K. Identification of clonogenic common lymphoid progenitors in mouse bone marrow. Cell 91, 661–672 (1997).
Adolfsson, J. et al. Identification of Flt3+ lympho-myeloid stem cells lacking erythro-megakaryocytic potential a revised road map for adult blood lineage commitment. Cell 121, 295–306 (2005).
Orkin, S.H. & Zon, L.I. Hematopoiesis: an evolving paradigm for stem cell biology. Cell 132, 631–644 (2008).
Luc, S., Buza-Vidas, N. & Jacobsen, S.E. Delineating the cellular pathways of hematopoietic lineage commitment. Semin. Immunol. 20, 213–220 (2008).
Doulatov, S. et al. Revised map of the human progenitor hierarchy shows the origin of macrophages and dendritic cells in early lymphoid development. Nat. Immunol. 11, 585–593 (2010).
Goardon, N. et al. Coexistence of LMPP-like and GMP-like leukemia stem cells in acute myeloid leukemia. Cancer Cell 19, 138–152 (2011).
Taub, D.D. & Longo, D.L. Insights into thymic aging and regeneration. Immunol. Rev. 205, 72–93 (2005).
Ceredig, R., Bosco, N. & Rolink, A.G. The B lineage potential of thymus settling progenitors is critically dependent on mouse age. Eur. J. Immunol. 37, 830–837 (2007).
Sambandam, A. et al. Notch signaling controls the generation and differentiation of early T lineage progenitors. Nat. Immunol. 6, 663–670 (2005).
Radtke, F. et al. Deficient T cell fate specification in mice with an induced inactivation of Notch1. Immunity 10, 547–558 (1999).
Feyerabend, T.B. et al. Deletion of Notch1 converts pro-T cells to dendritic cells and promotes thymic B cells by cell-extrinsic and cell-intrinsic mechanisms. Immunity 30, 67–79 (2009).
Hobeika, E. et al. Testing gene function early in the B cell lineage in mb1-cre mice. Proc. Natl. Acad. Sci. USA 103, 13789–13794 (2006).
Srinivas, S. et al. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus. BMC Dev. Biol. 1, 4 (2001).
Campbell, K.J. et al. Elevated Mcl-1 perturbs lymphopoiesis, promotes transformation of hematopoietic stem/progenitor cells, and enhances drug resistance. Blood 116, 3197–3207 (2010).
Kiel, M.J. et al. SLAM family receptors distinguish hematopoietic stem and progenitor cells and reveal endothelial niches for stem cells. Cell 121, 1109–1121 (2005).
Balazs, A.B., Fabian, A.J., Esmon, C.T. & Mulligan, R.C. Endothelial protein C receptor (CD201) explicitly identifies hematopoietic stem cells in murine bone marrow. Blood 107, 2317–2321 (2006).
Solar, G.P. et al. Role of c-mpl in early hematopoiesis. Blood 92, 4–10 (1998).
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).
Zlotoff, D.A. et al. CCR7 and CCR9 together recruit hematopoietic progenitors to the adult thymus. Blood 115, 1897–1905 (2010).
Krueger, A., Willenzon, S., Lyszkiewicz, M., Kremmer, E. & Forster, R. CC chemokine receptor (CCR) 7 and 9 double-deficient hematopoietic progenitors are severely impaired in seeding the adult thymus. Blood 115, 1906–1912 (2010).
Luc, S. et al. Downregulation of Mpl marks the transition to lymphoid-primed multipotent progenitors with gradual loss of granulocyte-monocyte potential. Blood 111, 3424–3434 (2008).
Godfrey, D.I., Kennedy, J., Suda, T. & Zlotnik, A. A developmental pathway involving four phenotypically and functionally distinct subsets of CD3−CD4−CD8− triple-negative adult mouse thymocytes defined by CD44 and CD25 expression. J. Immunol. 150, 4244–4252 (1993).
Mansson, R. et al. Molecular evidence for hierarchical transcriptional lineage priming in fetal and adult stem cells and multipotent progenitors. Immunity 26, 407–419 (2007).
Ehrlich, L.I., Serwold, T. & Weissman, I.L. In vitro assays misrepresent in vivo lineage potentials of murine lymphoid progenitors. Blood 117, 2618–2624 (2011).
Inlay, M.A. et al. Ly6d marks the earliest stage of B-cell specification and identifies the branchpoint between B-cell and T-cell development. Genes Dev. 23, 2376–2381 (2009).
Pronk, C.J.H. et al. Elucidation of the phenotypic, functional, and molecular topography of a myeloerythroid progenitor cell hierarchy. Cell Stem Cell 1, 428–442 (2007).
David-Fung, E.S. et al. Transcription factor expression dynamics of early T-lymphocyte specification and commitment. Dev. Biol. 325, 444–467 (2009).
Wendorff, A.A. et al. Hes1 is a critical but context-dependent mediator of canonical Notch signaling in lymphocyte development and transformation. Immunity 33, 671–684 (2010).
Petrie, H.T. & Kincade, P.W. Many roads, one destination for T cell progenitors. J. Exp. Med. 202, 11–13 (2005).
Chi, A.W. et al. Identification of Flt3CD150 myeloid progenitors in adult mouse bone marrow that harbor T lymphoid developmental potential. Blood 118, 2723–2732 (2011).
Schlenner, S.M. et al. Fate mapping reveals separate origins of T cells and myeloid lineages in the thymus. Immunity 32, 426–436 (2010).
Coustan-Smith, E. et al. Early T-cell precursor leukaemia: a subtype of very high-risk acute lymphoblastic leukaemia. Lancet Oncol. 10, 147–156 (2009).
Metzler, M. et al. A conditional model of MLL-AF4 B-cell tumourigenesis using invertor technology. Oncogene 25, 3093–3103 (2006).
Faust, N., Varas, F., Kelly, L.M., Heck, S. & Graf, T. Insertion of enhanced green fluorescent protein into the lysozyme gene creates mice with green fluorescent granulocytes and macrophages. Blood 96, 719–726 (2000).
Tehranchi, R. et al. Persistent malignant stem cells in del(5q) myelodysplasia in remission. N. Engl. J. Med. 363, 1025–1037 (2010).
Acknowledgements
We thank N. Sakaguchi (Kumamoto University) for mice with Rag1-driven GFP expression; T. Graf (Center for Genomic Regulation) for mice with expression of enhanced GFP (eGFP) driven by the gene encoding lysozyme M4; S. Cory (Walter and Eliza Hall Institute of Medical Research) for vavP-Mcl1–trangenic mice; M. Reth (Max Planck Institute of Immunobiology) for Cd79atm1(cre)Reth mice; S. Srinivas (University of Oxford) for mice expressing enhanced yellow fluorescent protein from the Rosa26 locus; A. Cumano (Institut Pasteur) for OP9 and OP9-DL1 stromal cells; Biomedical Services at Oxford University for animal support; S. Clark, T. Furey and B. Wu for technical assistance; and E. Zuo and M. Eckart at the Stanford Protein and Nucleic Acid Facility for gene array services. Supported by the Medical Research Council, UK (H4RPLK0 to S.E.W.J. and EU-FP7 EuroSyStem Integrated projects to S.E.W.J., C.B. and A.F.), Leukaemia and Lymphoma Research (C.B., A.F. and A.J.M.), the Crafoord Foundation (A.H.), The George Danielsson Foundation (A.H.) and Swedish Society for Medicine and Swedish Cancer Foundation (A.H.).
Author information
Authors and Affiliations
Contributions
S.E.W.J. and S.L. designed and conceived of the overall research, analyzed the data and wrote the manuscript, which was subsequently reviewed and approved by all authors; J.B. processed RNA samples; I.C.M. and S.S. analyzed the microarray data; A.J.M., D.A. and A.H. did quantitative and single-cell PCR; A.J.M., S.M. and K.A. did morphology analyses; H.F., S.L. and M.L. sorted cells by flow cytometry; S.L., M.L., T.B.-J., S.D., N.B.-V., H.B., T.C.L., A.D. and S.J.L. contributed to flow cytometry and in vitro culture experiments; S.L., S.D., N.B.-V., P.S.W., T.C.L. and H.B. did in vivo transplantations; T.E. provided assistance in the design and analysis of microarray experiments; C.B., A.F., R.P., M.d.B., I.G. and T.M. contributed advice and input on experimental design; and C.N., A.S.-P. and C.C. generated and provided input on studies of mice expressing eGFP driven by Vwf (encoding the von Willebrand factor homolog).
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Supplementary information
Supplementary Text and Figures
Supplementary Figures 1–9, Tables 1–5 and Note (PDF 2354 kb)
Rights and permissions
About this article
Cite this article
Luc, S., Luis, T., Boukarabila, H. et al. The earliest thymic T cell progenitors sustain B cell and myeloid lineage potential. Nat Immunol 13, 412–419 (2012). https://doi.org/10.1038/ni.2255
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/ni.2255
- Springer Nature America, Inc.
This article is cited by
-
Generation and clinical potential of functional T lymphocytes from gene-edited pluripotent stem cells
Experimental Hematology & Oncology (2022)
-
Acetylation licenses Th1 cell polarization to constrain Listeria monocytogenes infection
Cell Death & Differentiation (2022)
-
Differentiation Epitopes Define Hematopoietic Stem Cells and Change with Cell Cycle Passage
Stem Cell Reviews and Reports (2022)
-
A transcriptomic continuum of differentiation arrest identifies myeloid interface acute leukemias with poor prognosis
Leukemia (2021)
-
An Mb1-Cre-driven oncogenic Kras mutation results in a mouse model of T-acute lymphoblastic leukemia/lymphoma with short latency and high penetrance
Leukemia (2021)