Epitope mapping of human monoclonal antibodies to HLA-B27 by using natural and mutated antigenic variants.
The epitopes defined by three human monoclonal antibodies (mAbs) (Tr3B6, TrCG10, TrBH12) against HLA-B27 have been mapped by flow cytometry. For this purpose we used murine transfected cells expressing at their surface hybrid antigens between HLA-B7 and -B27 and, in addition, Epstein-Barr virus cell lines expressing the six HLA-B27 alleles B*2701 to B*2706. The results indicated that the mAbs are domain specific. TrBH12 recognizes the first external (alpha-1) domain. Residues critical for the TrBH12 epitope are located in the alpha-1 helix and include the polypeptide stretch 63-76 plus a critical amino acid at position 77. Tr3B6 binds the second external (alpha-2) domain, and one mutation (VAL152----GLU152) destroyed its epitope. TrCG10 also binds the alpha-2 domain.
Weyl
D
D
INSERM U. 283, Hôpital Cochin, Paris, France.
Hansen
T
T
Deschamps
Y
Y
Hannestad
K
K
Toubert
A
A
D000483:Alleles; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000940:Antigenic Variation / Q000276:immunology*; D000939:Epitopes / Q000276:immunology*; D005260:Female; D005434:Flow Cytometry; D015870:Gene Expression; D015796:HLA-B27 Antigen / Q000276:immunology*; D015793:HLA-B7 Antigen / Q000235:genetics; D004854:Herpesvirus 4, Human / Q000235:genetics; D006801:Humans; D051379:Mice; D016254:Mutagenesis, Insertional; D014162:Transfection; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015796:HLA-B27 Antigen; D015793:HLA-B7 Antigen