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


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 CD4loCD8CD25CD44+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+CD19CD4Thy1.1 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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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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Correspondence to Harald von Boehmer.

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