A transgenic reporter mouse strain, which expressed the human CD25 (hCD25) surface marker as a reporter under the control of the pre-T cell receptor α(pTα) promoter, was used to identify lymphoid precursors that expressed pTα intracellularly. The hCD25 reporter marked intra- and extrathymic precursors of lymphocytes but not myeloid cells. The earliest intrathymic precursors were CD4loCD8−CD25−CD44+c-Kit+ cells that expressed elevated levels of Notch-1 mRNA. Clonogenic assays showed that the extrathymic precursors were common lymphoid progenitors (CLPs) that included CD19−, B220+, Thy1+ and CD4+ cells. Thus, the pTα reporter can be used to trace lymphopoiesis between CLPs and αβ T cells. The slower extinction of the hCD25 reporter compared to pTα enabled us to define points at which pTα− lineages branched off.
Access optionsAccess options
Subscribe to Journal
Get full journal access for 1 year
only $18.75 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Rent or Buy article
Get time limited or full article access on ReadCube.
All prices are NET prices.
Spangrude, G. J., Heimfeld, S. & Weissman, I. L. Purification and characterization of mouse hematopoietic stem cells. Science 241, 58–62 (1988).
Morrison, S. J. & Weissman, I. L. The long-term repopulating subset of hematopoietic stem cells is deterministic and isolatable by phenotype. Immunity 1, 661–673 (1994).
Weissman, I. L., Anderson, D. J. & Gage, F. Stem and progenitor cells: origins, phenotypes, lineage commitments, and transdifferentiations. Annu. Rev. Cell Dev. Biol. 17, 387–403 (2001).
Adolfsson, J. et al. Upregulation of Flt3 expression within the bone marrow Lin−Sca1+c-kit+ stem cell compartment is accompanied by loss of self-renewal capacity. Immunity 15, 659–669 (2001).
Kondo, M., Weissman, I. L. & Akashi, K. Identification of clonogenic common lymphoid progenitors in mouse bone marrow. Cell 91, 661–672 (1997).
Fisher, R. C. & Scott, E. W. Role of PU.1 in hematopoiesis. Stem Cells 16, 25–37 (1998).
Urbanek, P., Wang, Z. Q., Fetka, I., Wagner, E. F. & Busslinger, M. Complete block of early B cell differentiation and altered patterning of the posterior midbrain in mice lacking Pax5/BSAP. Cell 79, 901–912 (1994).
O'Riordan, M. & Grosschedl, R. Coordinate regulation of B cell differentiation by the transcription factors EBF and E2A. Immunity 11, 21–31 (1999).
Wilson, A., MacDonald, H. R. & Radtke, F. Notch 1-deficient common lymphoid precursors adopt a B cell fate in the thymus. J. Exp. Med. 194, 1003–1012 (2001).
Radtke, F. et al. Deficient T cell fate specification in mice with an induced inactivation of Notch1. Immunity 10, 547–558 (1999).
Allman, D. et al. Separation of Notch1 promoted lineage commitment and expansion/transformation in developing T cells. J. Exp. Med. 194, 99–106 (2001).
Pui, J. C. et al. Notch1 expression in early lymphopoiesis influences B versus T lineage determination. Immunity 11, 299–308 (1999).
Jaleco, A. C. et al. Differential effects of Notch ligands Delta-1 and Jagged-1 in human lymphoid differentiation. J. Exp. Med. 194, 991–1002 (2001).
Groettrup, M. et al. A novel disulfide-linked heterodimer on pre-T cells consists of the T cell receptor β chain and a 33 kd glycoprotein. Cell 75, 283–294 (1993).
Saint-Ruf, C. et al. Analysis and expression of a cloned pre-T cell receptor gene. Science 266, 1208–1212 (1994).
Deftos, M. L., Huang, E., Ojala, E. W., Forbush, K. A. & Bevan, M. J. Notch1 signaling promotes the maturation of CD4 and CD8 SP thymocytes. Immunity 13, 73–84 (2000).
Reizis, B. & Leder, P. Direct induction of T lymphocyte specific gene expression by the mammalian Notch signaling pathway. Genes Dev. 16, 295–300 (2002).
Bruno, L., Rocha, B., Rolink, A., von Boehmer, H. & Rodewald, H. R. Intra- and extra-thymic expression of the pre-T cell receptor α gene. Eur. J. Immunol. 25, 1877–1882 (1995).
Martensson, I. L., Melchers, F. & Winkler, T. H. A transgenic marker for mouse B lymphoid precursors. J. Exp. Med. 185, 653–661 (1997).
Monroe, R. J. et al. RAG2:GFP knockin mice reveal novel aspects of RAG2 expression in primary and peripheral lymphoid tissues. Immunity 11, 201–212 (1999).
Yu, W. et al. Continued RAG expression in late stages of B cell development and no apparent re-induction after immunization. Nature 400, 682–687 (1999).
Reizis, B. & Leder, P. The upstream enhancer is necessary and sufficient for the expression of the Pre-T cell receptor α gene in immature T lymphocytes. J. Exp. Med. 194, 979–990 (2001).
Reizis, B. & Leder, P. Expression of the mouse pre-T cell receptor α gene is controlled by an upstream region containing a transcriptional enhancer. J. Exp. Med. 189, 1669–1678 (1999).
Nishi, M., Ishida, Y. & Honjo, T. Expression of functional interleukin-2 receptors in human light chain/Tac transgenic mice. Nature 331, 267–269 (1988).
Shortman, K. & Wu, L. Early T lymphocyte progenitors. Annu. Rev. Immunol. 14, 29–47 (1996).
Wu, L., Antica, M., Johnson, G. R., Scollay, R. & Shortman, K. Developmental potential of the earliest precursor cells from the adult mouse thymus. J. Exp. Med. 174, 1617–1627 (1991).
Matsuzaki, Y. et al. Characterization of c-kit positive intrathymic stem cells that are restricted to lymphoid differentiation. J. Exp. Med. 178, 1283–1292 (1993).
Aifantis, I. et al. On the role of the pre-T cell receptor in αβ versus γδ T lineage commitment. Immunity 9, 649–655 (1998).
Yu, W. et al. Coordinate regulation of RAG1 and RAG2 by cell type-specific DNA elements 5′ of RAG2. Science 285, 1080–1084 (1999).
Hardy, R. R. & Hayakawa, K. B cell development pathways. Annu. Rev. Immunol. 19, 595–621 (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).
Res, P. C., Couwenberg, F., Vyth-Dreese, F. A. & Spits, H. Expression of pTα mRNA in a committed dendritic cell precursor in the human thymus. Blood 94, 2647–2657 (1999).
Ikawa, T., Kawamoto, H., Fujimoto, S. & Katsura, Y. Commitment of common T/Natural killer (NK) progenitors to unipotent T and NK progenitors in the murine fetal thymus revealed by a single progenitor assay. J. Exp. Med. 190, 1617–1626 (1999).
Kawamoto, H., Ohmura, K., Fujimoto, S. & Katsura, Y. Emergence of T cell progenitors without B cell or myeloid differentiation potential at the earliest stage of hematopoiesis in the murine fetal liver. J. Immunol. 162, 2725–2731 (1999).
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).
Bruno, L., Seidl, T. & Lanzavecchia, A. Mouse pre-immunocytes as non-proliferating multipotent precursors of macrophages, interferon-producing cells, CD8a+ and CD8a- dendritic cells. Eur. J. Immunol. 31, 3403–3412 (2001).
Asselin-Paturel, C. et al. Mouse type I IFN-producing cells are immature APCs with plasmacytoid morphology. Nature Immunol. 2, 1144–1150 (2001).
Wu, L. et al. CD4 expressed on earliest T-lineage precursor cells in the adult murine thymus. Nature 349, 71–74 (1991).
Bruno, L., Res, P., Dessing, M., Cella, M. & Spits, H. Identification of a committed T cell precursor population in adult human peripheral blood. J. Exp. Med. 185, 875–884 (1997).
Georgopoulos, K. Haematopoietic cell fate decisions, chromatin regulation and Ikaros. Nature Rev. Immunol. 2, 162–174 (2002).
Dejbakhsh-Jones, S., Jerabek, L., Weissman, I. L. & Strober, S. Extrathymic maturation of β T cells from hemopoietic stem cells. J. Immunol. 155, 3338–3344 (1995).
Douagi, I. I., Andre, I. I., Ferraz, J. C. & Cumano, A. Characterization of T cell precursor activity in the murine fetal thymus: evidence for an input of T cell precursors between days 12 and 14 of gestation. Eur. J. Immunol. 30, 2201–2210 (2000).
Supported by NIH grants (R01A147281 and R01A145846 to H. v. B.); the “Eugenia Spanopoulou” Irvington Award (to I. A.), the Division of Medical Sciences of the Harvard Medical School (C. M.); and the Cancer Research Institute (B.R.). We thank K. Akashi and P. Kincade for advice and encouragement; A. Harrington for oocyte injections; T. Honjo and Y. Ishida for the hCD25 minigene; and M. Handley and H. Levine for cell sorting. Supported by the Albert J. Ryan Foundation (to C.M.).
The authors declare no competing financial interests.
About this article
The Journal of Immunology (2015)
Annual Review of Immunology (2015)
Nature Immunology (2014)
In vivo fate mapping identifies pre-TCRα expression as an intra- and extrathymic, but not prethymic, marker of T lymphopoiesis
The Journal of Experimental Medicine (2013)