Characterization and quantitation of peptide-MHC complexes produced from hen egg lysozyme using a monoclonal antibody.
Here we describe generation of Aw3.18, a monoclonal antibody that recognizes peptide residues 48-62 of hen egg lysozyme (HEL) bound to the MHC class II molecule I-Ak. Epitope mapping revealed that Aw3.18 detects a change in the solvent-exposed surface of this peptide-MHC complex upon substitution of the peptide side chain at position P1. Furthermore, Aw3.18 blocked recognition by some, but not all, of the HEL 48-62-reactive T cell hybridomas tested, suggesting a heterogeneity in the T cell response toward this complex. Finally, using Aw3.18, it was possible to determine the fraction of I-Ak molecules loaded with 48-62 peptide after culture of an antigen-presenting cell in medium containing HEL.
Dadaglio
G
G
Center for Immunology, Department of Pathology, Washington Unviversity School of Medicine, St. Louis, Missouri 63110, USA.
Nelson
C A
CA
Deck
M B
MB
Petzold
S J
SJ
Unanue
E R
ER
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000938:Antigen-Presenting Cells / Q000276:immunology; D001402:B-Lymphocytes / Q000276:immunology; D001666:Binding Sites, Antibody; D001667:Binding, Competitive; D000949:Histocompatibility Antigens Class II / Q000276:immunology*; D046911:Macromolecular Substances; D051379:Mice; D008969:Molecular Sequence Data; D009113:Muramidase / Q000276:immunology; D010455:Peptides / Q000276:immunology*; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D013601:T-Lymphocytes / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000949:Histocompatibility Antigens Class II; D046911:Macromolecular Substances; D010455:Peptides; D009113:Muramidase