Immunobiological analysis of TCR single-chain transgenic mice reveals new possibilities for interaction between CDR3alpha and an antigenic peptide bound to MHC class I.
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.
Zhang
W
W
Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Honda
S
S
Wang
F
F
DiLorenzo
T P
TP
Kalergis
A M
AM
Ostrov
D A
DA
Nathenson
S G
SG
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
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