Epitope mapping of HLA-B27 and HLA-B7 antigens by using intradomain recombinants.
To study the HLA-B7 and HLA-B27 antigenic determinants, hybrid genes between these two alleles were constructed by in vivo recombination in Escherichia coli. After transfection of these genes into P815 (high transfection efficiency recipient) murine cells, the bindings of Bw6, HLA-B7, and HLA-B27 allele-specific mAb were studied, as well as that of human anti-HLA-B7 and anti-HLA-B27 monospecific alloantisera. Most of the HLA-B7 antigenic determinants were assigned to the first external domain of the molecule. Four different epitopic areas could be defined: the Bw6 epitope was associated with residues 82 and 83; the BB7.1 epitope to amino acids 63, 67, and 70; the MB40.2 and MB40.3 epitope to amino acid sequence 177-180, and human alloantisera identified as an epitope associated with residue 9. HLA-B27 antigenicity studied by TM-1 mAb was found to involve residues 77 and 80 in the alpha-1 domain. Results obtained with human monospecific alloantisera allowed the definition of an additional allospecific site associated with the NH2 terminal part on the alpha-1 domain of HLA-B27. Epitope mapping fits with data obtained by sequence comparisons and is discussed with reference to the crystallographic three-dimensional structure of the HLA-A2 molecule.
Toubert
A
A
INSERM, CNRS, Marseille-Luminy, France.
Raffoux
C
C
Boretto
J
J
Sire
J
J
Sodoyer
R
R
Thurau
S R
SR
Amor
B
B
Colombani
J
J
Lemonnier
F A
FA
Jordan
B R
BR
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D003601:Cytotoxicity Tests, Immunologic; D000939:Epitopes / Q000235:genetics* / Q000276:immunology; D005260:Female; D005813:Genes, Synthetic; D015235:HLA-B Antigens / Q000235:genetics* / Q000276:immunology; D015796:HLA-B27 Antigen; D015793:HLA-B7 Antigen; D006650:Histocompatibility Testing; D006801:Humans; D007518:Isoantibodies; D051379:Mice; D008969:Molecular Sequence Data; D010449:Peptide Mapping; D011995:Recombination, Genetic; D014162:Transfection
D000911:Antibodies, Monoclonal; D000939:Epitopes; D015235:HLA-B Antigens; D015796:HLA-B27 Antigen; D015793:HLA-B7 Antigen; D007518:Isoantibodies