title: Immunobiological analysis of TCR single-chain transgenic mice reveals new possibilities for interaction between CDR3alpha and an antigenic peptide bound to MHC class I.
abstract: The interaction between TCRs and peptides presented by MHC molecules determines the specificity of the T cell-mediated immune response. To elucidate the biologically important structural features of this interaction, we generated TCR beta-chain transgenic mice using a TCR derived from a T cell clone specific for the immunodominant peptide of vesicular stomatitis virus (RGYVYQGL, VSV8) presented by H-2K(b). We immunized these mice with VSV8 or analogs substituted at TCR contact residues (positions 1, 4, and 6) and analyzed the CDR3alpha sequences of the elicited T cells. In VSV8-specific CTLs, we observed a highly conserved residue at position 93 of CDR3alpha and preferred Jalpha usage, indicating that multiple residues of CDR3alpha are critical for recognition of the peptide. Certain substitutions at peptide position 4 induced changes at position 93 and in Jalpha usage, suggesting a potential interaction between CDR3alpha and position 4. Cross-reactivity data revealed the foremost importance of the Jalpha region in determining Ag specificity. Surprisingly, substitution at position 6 of VSV8 to a negatively charged residue induced a change at position 93 of CDR3alpha to a positively charged residue, suggesting that CDR3alpha may interact with position 6 in certain circumstances. Analogous interactions between the TCR alpha-chain and residues in the C-terminal half of the peptide have not yet been revealed by the limited number of TCR/peptide-MHC crystal structures reported to date. The transgenic mouse approach allows hundreds of TCR/peptide-MHC interactions to be examined comparatively easily, thus permitting a wide-ranging analysis of the possibilities for Ag recognition in vivo.
authors.affiliation: Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
authors.forename: W
authors.initials: W
authors.lastname: Zhang
authors.forename: S
authors.initials: S
authors.lastname: Honda
authors.forename: F
authors.initials: F
authors.lastname: Wang
authors.forename: T P
authors.initials: TP
authors.lastname: DiLorenzo
authors.forename: A M
authors.initials: AM
authors.lastname: Kalergis
authors.forename: D A
authors.initials: DA
authors.lastname: Ostrov
authors.forename: S G
authors.initials: SG
authors.lastname: Nathenson
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017951:Antigen Presentation; D000956:Antigens, Viral / Q000276:immunology; D022801:Complementarity Determining Regions / Q000378:metabolism*; D017124:Conserved Sequence; D019674:Genes, T-Cell Receptor beta; D006183:H-2 Antigens / Q000378:metabolism*; D016056:Immunodominant Epitopes / Q000276:immunology; D051379:Mice; D008822:Mice, Transgenic; D008958:Models, Molecular; D009842:Oligopeptides / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology; D011485:Protein Binding; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000235:genetics / Q000378:metabolism*; D013602:T-Lymphocytes, Cytotoxic; D014721:Vesicular stomatitis Indiana virus / Q000276:immunology
chemical_list: D000956:Antigens, Viral; D022801:Complementarity Determining Regions; D006183:H-2 Antigens; C032876:H-2Kb protein, mouse; D016056:Immunodominant Epitopes; D009842:Oligopeptides; D010446:Peptide Fragments; D016693:Receptors, Antigen, T-Cell, alpha-beta