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Evidence for Two Subgroups of CD4^-CD8^- NKT Cells with Distinct TCRß Repertoires and Differential Distribution in Lymphoid Tissues¹

Irina Apostolou^2,*, Ana Cumano, Gabriel Gachelin^* and Philippe Kourilsky^*

^* Unité de Biologie Moléculaire du Gène, Institut National de la Santé et de la Recherche Médicale Unité 277, and Institut Pasteur, Paris, France; and the Unité du Développement des Lymphocytes, Institut Pasteur, Paris, France

NKT cells are a subset of T lymphocytes that is mainly restricted by the nonclassical MHC class I molecule, CD1d, and that includes several subpopulations, in particular CD4⁺ and CD4^-CD8^- (DN) cells. In the mouse, differential distribution of these subpopulations as well as heterogeneity in the expression of various markers as a function of tissue localization have been reported. We have thus undertaken a detailed study of the DN NKT cell subpopulation. With a highly sensitive semiquantitative RT-PCR technique, its TCR repertoire was characterized in various tissues. We found that mouse DN NKT cells are a variable mixture of two subgroups, one bearing the invariant V14 chain paired to rearranged Vß2, Vß7, Vß8.1, Vß8.2, or Vß8.3 ß-chains and the other exhibiting unskewed - and ß-chains. The proportion of these subgroups varies from about 100:0 in thymus, 80:20 in liver, and 50:50 in spleen to 20:80% in bone marrow, respectively. Finally, further heterogeneity in the tissue-derived DN NKT cells was discovered by sequencing extensively Vß8.2-Jß2.5 rearrangements in individual mice. Despite a few recurrences in TCR sequences, we found that each population exhibits its own and broad TCRß diversity.

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