Identification of residues involved in polymorphic antibody binding epitopes on HLA-DR molecules.
Based on previous studies it was predicted that amino acids 4 or 25 of the DR4 beta 1 and DR7 beta 1 chains are involved in polymorphic antibody binding epitopes on DR4 or DR7 molecules. These predictions were tested by analyzing monoclonal antibody (mAb) binding to transfectants expressing mutant DR4 beta 1 or DR7 beta 1 chains with single amino acid substitutions at positions 4 or 25. Antibody binding to transfectants expressing additional DR4/7 beta 1 hybrids was also analyzed to assess further the contributions of four segments of the DR4 beta 1 or DR7 beta 1 chains: amino acids 1-20, 21-40, 41-97, and the beta 2 domain. Single amino acid substitutions at positions 4 and 25 of the DR4 beta 1 chain or DR7 beta 1 chain eliminate binding of several mAb to DR4 or DR7 molecules, documenting that these residues are involved in antibody epitopes. However, the data with the hybrid DR4/7 beta 1 chains indicate that some of these epitopes require contributions from both segments 1-20 and 21-40 of these DR beta chains, whereas other epitopes can be generated by placing the appropriate segment in the context of the other DR beta chain. In addition, the data with other mAb indicate that their epitopes are determined primarily by sequences within the 41-97 segment or in the beta 2 domain.
Department of Veterans Affairs Medical Center, Iowa City, Iowa.
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Fu
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Yu
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Alber
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Benson
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Watts
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Nordwig
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Johnson
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Knowles
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Karr
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D003001:Cloning, Molecular; D000939:Epitopes / Q000235:genetics / Q000276:immunology*; D005455:Fluorescent Antibody Technique; D006684:HLA-DR Antigens / Q000235:genetics / Q000276:immunology*; D007739:L Cells; D051379:Mice; D016297:Mutagenesis, Site-Directed; D009693:Nucleic Acid Hybridization; D011110:Polymorphism, Genetic; D014162:Transfection
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000939:Epitopes; D006684:HLA-DR Antigens