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Yoshinori Fukui^*,, Osamu Hashimoto^*, Ayumi Inayoshi^*, Takahiro Gyotoku^*, Tetsuro Sano^*, Takahiro Koga^*, Toshifumi Gushima^*, and Takehiko Sasazuki^*,

From the ^* Department of Genetics, Medical Institute of Bioregulation, Kyushu University, and Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Corporation, Fukuoka 812-8582, Japan

The T cell repertoire is shaped by positive and negative selection of thymocytes through the interaction of /β-T cell receptors (TCR) with self-peptides bound to self-major histocompatibility complex (MHC) molecules. However, the involvement of specific TCR-peptide contacts in positive selection remains unclear. By fixing TCR-β chains with a single rearranged TCR-β irrelevant to the selecting ligand, we show here that T cells selected to mature on a single MHC–peptide complex express highly restricted TCR- chains in terms of V usage and amino acid residue of their CDR3 loops, whereas such restriction was not observed with those selected by the same MHC with diverse sets of self-peptides including this peptide. Thus, we visualized the TCR structure required to survive positive selection directed by this single ligand. Our findings provide definitive evidence that specific recognition of self-peptides by TCR could be involved in positive selection of thymocytes.

Key Words: positive selection • single major histocompatibility complex–peptide complex • single rearranged T cell receptor β chain • T cell repertoire • transgenic knockout mice

Abbreviations used: β2^0/0, mice lacking β2-microglobulin; B2L or B2H, mouse lines expressing I-Aβ^b chain covalently bound to E52-68; B6, C57BL/6 mice; CLIP, invariant chain–derived class II–associated peptide; DKO, mice lacking endogenous I-Aβ^b and invariant chains; E52-68, E-derived peptide; E-B6, C57BL/6 mice transgenic for E; H-2M^0/0, mice lacking H-2M expression; TKO, mice lacking endogenous I-Aβ^b, invariant chain, and β2-microglobulin; TKO/2B4β, B2L TKO/2B4β, β2^0/0/2B4β, E-B6/2B4β; TKO, B2L TKO, β2^0/0 and E-B6 mice expressing 2B4 TCR-β chain.

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