The recognition of gammadelta TCR to protein antigen does not depend on the hydrophobic I97 residue of CDR3delta.
The crystal structure analysis demonstrated that the hydrophobic amino acid residue (isolecuine/leucine/valine) at conserved position 97 of Vdelta2 TCR plays an important role in recognizing the non-peptide antigen. But its importance to protein antigen remains unclear until now. In the present study, we focus on the role of hydrophobic amino acid residue at conserved position 97 of Vdelta2 TCR in complementarity determining region (CDR)3delta-mediated binding to protein antigen. We employed CDR3delta peptide and membrane-engineered gammadelta TCR as detecting molecules with mutated 97 hydrophobic amino acid residue in CDR3delta (nominated as OT10), a Vdelta2 CDR3 sequence derived from tumor infiltrating lymphocytes in ovarian epithelial carcinoma (OEC). Binding assays revealed that OT10 peptide and membrane-engineered gammadelta TCR (gammadelta TCR transfected cells with OT10 sequence) could bind specifically ovarian tumor cell line (SKOV3). The mutant analysis indicated that any amino acid substitution at position deltaI97 could abolish the response of the transfected cells to iso-butylamine, a known non-peptide antigen of gammadelta T cells. But amino acid substitution of isoleucine at position delta97 did not change the responsiveness of gammadelta TCR transfected cell to protein antigen. Our data suggested that a mechanism other than non-peptide antigen might mediate the recognition of Vdelta2gammadelta T cells for protein antigen. This finding may provide a possibility that gammadelta TCR recognize different ligands in diversity manners.
Xi
Xueyan
X
Department of Immunology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and School of Basic Medicine, Peking Union Medical College, 5 Dong Dan San Tiao, Beijing 100005, China.
Cui
Lianxian
L
He
Wei
W
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000941:Antigens / Q000276:immunology*; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D003001:Cloning, Molecular; D022801:Complementarity Determining Regions / Q000235:genetics / Q000276:immunology*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007532:Isoleucine / Q000235:genetics / Q000276:immunology*; D019169:Jurkat Cells; D008969:Molecular Sequence Data; D010455:Peptides / Q000276:immunology; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000235:genetics / Q000276:immunology*; D013601:T-Lymphocytes / Q000276:immunology
D000941:Antigens; D022801:Complementarity Determining Regions; D010455:Peptides; D016692:Receptors, Antigen, T-Cell, gamma-delta; C521523:T-cell receptor Vdelta2, human; D007532:Isoleucine