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Tracing lymphopoiesis with the aid of a pTα-controlled reporter gene

Nature Immunology volume 3, pages 489–496 (2002)Cite this article

Abstract

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.

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Figure 1: Characterization of hCD25 transgene expression in the thymus and periphery.
Figure 2: Thymic γδ cells originate from a pTα-expressing precursor.
Figure 3: Phenotype of BM hCD25+ cells.
Figure 4: In vitro culture of the hCD25+CD19−CD4−Thy1.1− population.
Figure 5: In vivo reconstitution potential of BM hCD25+ cells.
Figure 6: Clonogenic and small subpopulation developmental potential of hCD25+CD19− BM progenitors.
Figure 7: Expression of lineage-specific genes in pTα-expressing populations.

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Acknowledgements

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.).

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Author notes

  1. Fotini Gounari, Iannis Aifantis and Colin Martin: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pathology, Harvard Medical School, Dana-Farber Cancer Institute, Boston, 02115, MA, USA

    Fotini Gounari, Iannis Aifantis, Colin Martin, Sonja Hoeflinger & Harald von Boehmer

  2. Department of Immunology, Medical Faculty/University Clinics Ulm, D-89070, Ulm, Germany

    Hans-Joerg Fehling

  3. Department of Genetics and Howard Hughes Medical Institute, Harvard Medical School, Boston, 02115, MA, USA

    Philip Leder & Boris Reizis

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Correspondence to Harald von Boehmer.

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Gounari, F., Aifantis, I., Martin, C. et al. Tracing lymphopoiesis with the aid of a pTα-controlled reporter gene. Nat Immunol 3, 489–496 (2002). https://doi.org/10.1038/ni778

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