Abstract
The checkpoint in γδ cell development that controls successful T cell receptor (TCR) gene rearrangements remains poorly characterized. Using mice expressing a reporter gene 'knocked into' the Tcrd constant region gene, we have characterized many of the events that mark the life of early γδ cells in the adult thymus. We identify the developmental stage during which the Tcrd locus 'opens' in early T cell progenitors and show that a single checkpoint controls γδ cell development during the penultimate CD4−CD8− stage. Passage through this checkpoint required the assembly of γδ TCR heterodimers on the cell surface and signaling via the Lat adaptor protein. In addition, we show that γδ selection triggered a phase of sustained proliferation similar to that induced by the pre-TCR.
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Acknowledgements
We thank C.A. Stewart, D.J. Pennington (University of London, London, UK), W. Held (Ludwig Institute for Cancer Research, Lausanne, Switzerland) and R. Ceredig (Center for Biomedecine, Basel, Switzerland) for discussions; and P. Grenot, M. Barad, F. Danjan, M. Richelme, P. Perrin, C. Grégoire, M. Fallet, S. Sarazin and A. Gillet for advice. Supported by Centre National de la Recherche Scientifique (B.M.), Institut National de la Santé et de la Recherche Médicale (B.M.), Association pour la Recherche contre le Cancer (B.M.), Fondation pour la Recherche Médicale (B.M.), Ministère de l'Education Nationale et de la Recherche (Plate-forme RIO-MNG; B.M.), Agence National de Recherches (B.M.), the European Community (MUGEN Network of Excellence; B.M.) and a Marie Curie Intra-European Fellowship within the 6th European Community Framework Program (I.P.).
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All authors contributed to discussions of experimental design and data analysis; I.P. did all experimental studies unless otherwise indicated; A.S. provided technical assistance; A.K. helped design the 'knock-in' strategy; L.A. and M.M. generated the Janus kinase 3–deficient mice; and I.P. and B.M. wrote the manuscript.
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Supplementary information
Supplementary Fig. 1
Generation and identification of Tcrd-H2BEGFP mice. (PDF 2726 kb)
Supplementary Fig. 2
Schematic of rearrangement, transcription and translation of the targeted Tcrd gene. (PDF 669 kb)
Supplementary Fig. 3
Normal percentages and numbers of thymocyte subsets in lymphoid organs of Tcrd-H2BEGFP mice. (PDF 55 kb)
Supplementary Fig. 4
Comparable numbers of T cells, NK cells and NKT cells in lymphoid organs of WT and Tcrd-H2BEGFP mice. (PDF 17 kb)
Supplementary Fig. 5
Lineage-negative bone marrow cells from Tcrd-H2BEGFP mice efficiently generate γδ T cells during competitive reconstitution. (PDF 17 kb)
Supplementary Fig. 6
Tcrd transcription occurs in the absence of Jak3. (PDF 89 kb)
Supplementary Fig. 7
Turnover of EGFPhigh DN3 and EGFPhigh DN4 cells from Tcrd-H2BEGFP mice. (PDF 11 kb)
Supplementary Fig. 8
Earliest steps of αβ and γδ cell development in adult thymus. (PDF 21 kb)
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Prinz, I., Sansoni, A., Kissenpfennig, A. et al. Visualization of the earliest steps of γδ T cell development in the adult thymus. Nat Immunol 7, 995–1003 (2006). https://doi.org/10.1038/ni1371
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DOI: https://doi.org/10.1038/ni1371
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