The Journal of Experimental Medicine
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Published 11 February 2002. doi:10.1084/jem.20010798
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© Rockefeller University Press, 0022-1007/2002/2/437/ $5.00
The Journal of Experimental Medicine, Volume 195, Number 4, February 18, 2002 437-449

Original Article

 Characterization of T Cell Differentiation in the Murine Gut

Florence Lambolez1, Orly Azogui2, Anne-Marie Joret1, Corinne Garcia3, Harald von Boehmer4, James Di Santo5, Sophie Ezine1 and Benedita Rocha1

1 Institut National de la Santé et de la Recherche Médicale (INSERM) U345, Institut Necker, 75730 Paris, France
2 E9925, Institut Necker, 75730 Paris, France
3 U373, Institut Necker, 75730 Paris, France
4 Harvard Medical School, Dana-Farber Cancer Institute, Boston, MA 02115
5 Institut Pasteur, 75015 Paris, France

Address correspondence to Benedita Rocha, INSERM U345, Institut Necker, CHU Necker, 156, Rue de Vaugirard, 75730 Paris Cedex 15, France. Phone: 33-1-40-61-53-65; Fax: 33-1-40-61-55-80; E-mail: rocha{at}necker.fr

Gut intraepithelial CD8 T lymphocytes (T-IEL) are distinct from thymus-derived cells and are thought to derive locally from cryptopatch (CP) precursors. The intermediate stages of differentiation between CP and mature T-IEL were not identified, and the local differentiation process was not characterized. We identified and characterized six phenotypically distinct lineage-negative populations in the CP and the gut epithelium: (a) we determined the kinetics of their generation from bone marrow precursors; (b) we quantified CD3-{epsilon}, recombination activating gene (Rag)-1, and pre-T{alpha} mRNAs expression at single cell level; (c) we characterized TCR-ß, -{gamma}, and -{alpha} locus rearrangements; and (d) we studied the impact of different mutations on the local differentiation. These data allowed us to establish a sequence of T cell precursor differentiation in the gut. We also observed that the gut differentiation varied from that of the thymus by a very low frequency of pre-T{alpha} chain mRNA expression, a different kinetics of Rag-1 mRNA expression, and a much higher impact of CD3 {epsilon}/{delta} and pre-T{alpha} deficiencies. Finally, only 3% of CP cells were clearly involved in T cell differentiation, suggesting that these structures may have additional physiological roles in the gut.

Key Words: T cells • precursors • differentiation • pre-TCR{alpha} • CD3-{epsilon}


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