The Journal of Experimental Medicine
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© The Rockefeller University Press, 0022-1007/1998/6/1871/ $5.00
The Journal of Experimental Medicine, Volume 187, Number 11, June 1, 1998 1871-1883

Articles

Quantitative Analysis of the T Cell Repertoire Selected by a Single Peptide–Major Histocompatibility Complex

Laurent Gapin*, Yoshinori Fukui{ddagger}, Jean Kanellopoulos*, Tetsuro Sano{ddagger}, Armanda Casrouge*, Vanessa Malier*, Emmanuel Beaudoing§, Daniel Gautheret§, Jean-Michel Claverie§, Takehiko Sasazuki{ddagger}, and Philippe Kourilsky*

From the * Laboratoire de Biologie Moléculaire du Gène, Institut National de la Santé et de la Recherche Médicale U277 and Institut Pasteur, 75724 Paris, France; the {ddagger} Department of Genetics, Medical Institute of Bioregulation and Ministry of Education, Science, Sports and Culture, Japan Science and Technology Corporation, Fukuoka 812, Japan; and the § Information Genetique et Structurale, Centre National de la Recherche Scientifique, Institut de Biologie Structurale et Microbiologie, 13402 Marseille, France

The positive selection of CD4+ T cells requires the expression of major histocompatibility complex (MHC) class II molecules in the thymus, but the role of self-peptides complexed to class II molecules is still a matter of debate. Recently, it was observed that transgenic mice expressing a single peptide–MHC class II complex positively select significant numbers of diverse CD4+ T cells in the thymus. However, the number of selected T cell specificities has not been evaluated so far. Here, we have sequenced 700 junctional complementarity determining regions 3 (CDR3) from T cell receptors (TCRs) carrying Vβ11-Jβ1.1 or Vβ12-Jβ1.1 rearrangements. We found that a single peptide–MHC class II complex positively selects at least 105 different Vβ rearrangements. Our data yield a first evaluation of the size of the T cell repertoire. In addition, they provide evidence that the single E{alpha}52-68–I-Ab complex skews the amino acid frequency in the TCR CDR3 loop of positively selected T cells. A detailed analysis of CDR3 sequences indicates that a fraction of the β chain repertoire bears the imprint of the selecting self-peptide.

Key Words: thymus • major histocompatibility complex • T cell receptors • repertoire development • transgenic/knockout

Address correspondence to Philippe Kourilsky, Laboratoire de Biologie Moléculaire du Gène, INSERM U277 and Institut Pasteur, 28, rue du Docteur Roux, 75724 Paris cedex 15, France. Phone: 33-1-45-68-85-45; Fax: 33-1-45-68-85-48; E-mail: kourilsk{at}pasteur.fr

L. Gapin is supported by l'Association pour la Recherche contre le Cancer and Pasteur-Weizmann fellowships. This work was supported by grants from Institut National de la Santé et de la Recherche Médicale and Institut Pasteur.

L. Gapin's current address is La Jolla Institute for Allergy and Immunology, Division of Developmental Immunology, San Diego, CA 92121.

Abbreviations used: CLIP, class II–associated invariant chain peptide; Tg, transgenic mice.


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