A study of functionally active amino acids involved in the interaction of HLA-A2 or HLA-A3 molecules with cytolytic T lymphocytes.
A large series of HLA-A2/HLA-A3 recombinant genes were generated by using the in vivo recombination technique. These genes have each been modified in the last two-thirds of the third exon such that one or several HLA-A2-specific substitutions have been made in the HLA-A3 gene and vice versa. The recombinant genes were transfected into the murine cell line P815 and the transfectants were used as targets for a series of 20 human CTL lines or clones specific for HLA-A2 or HLA-A3, or restricted by HLA-A2 and specific for influenza A. Several patterns of anti-HLA-A2, anti-HLA-A3, and HLA-A2-restricted anti-influenza CTL activity were observed and when uncloned cell lines were studied, a progressive selection of some clones with a similar pattern of activity was regularly found. From the comparison of these different patterns the following conclusions can be drawn: 1) In most but not all cases both domains of the class I molecule were essential for CTL recognition, but residue 152 was critically important for the majority of CTL tested; 2) amino acids 114/116 were also critical in most cases, and their position close to amino acid 152 in the tertiary structure of the molecule may have some functional significance; and 3) amino acid 161, although highly conserved, plays an unexpected but very important role in CTL function.
Healy
F
F
Laboratoire d'Immunologie et de Virologie des Tumeurs, INSERM 152, Paris, France.
Sire
J
J
Gomard
E
E
Yssel
H
H
Jordan
B
B
Levy
J P
JP
D000596:Amino Acids / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000961:Antilymphocyte Serum / Q000276:immunology; D001667:Binding, Competitive; D001782:Blood Donors; D002460:Cell Line; D003601:Cytotoxicity Tests, Immunologic / Q000379:methods; D000939:Epitopes / Q000276:immunology; D015234:HLA-A Antigens / Q000032:analysis / Q000276:immunology*; D015789:HLA-A2 Antigen; D015791:HLA-A3 Antigen; D006801:Humans; D009980:Influenza A virus / Q000276:immunology; D051379:Mice; D010455:Peptides / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000961:Antilymphocyte Serum; D000939:Epitopes; D015234:HLA-A Antigens; D015789:HLA-A2 Antigen; D015791:HLA-A3 Antigen; D010455:Peptides