title: Monoclonal antibodies against Dermatophagoides group I allergens as pseudo-cystatins blocking the catalytic site of cysteine proteinases.
abstract: The enzyme allergens Der p I and Der f I produced by the house dust mites Dermatophagoides pteronyssinus and D. farinae display partial sequence homology with other members of the cysteine proteinase superfamily. We report that certain widely used mouse mAbs against these Group I allergens indeed crossreact with the plant enzymes papain, bromelain and ficin. The recognition sites of these anti Group I mAbs comprise conformational and thermolabile epitopes involved in molding the catalytic center of the proteinases. Thus, the mAbs inhibit the enzymatic hydrolysis of specific chromogenic substrates by the Group I allergens, while specific cysteine proteinase inhibitors abolish the recognition of the enzymes by the mAbs. Similarly, activation of the thiol-proteases with L-cysteine abrogates their binding in the two-site mAb system, indicating that the mAbs recognize a proenzyme conformational peptide epitope. It follows that mAb-based assays for mite Group I components can neither detect the allergens after inactivation, nor in their fully activated forms.
authors.affiliation: Research Laboratories CBF LETI, Tres Cantos, Madrid, Spain.
authors.forename: O
authors.initials: O
authors.lastname: Cambra
authors.forename: L
authors.initials: L
authors.lastname: Berrens
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D039741:Antigens, Dermatophagoides; D001963:Bromelains / Q000276:immunology; D003429:Cross Reactions; D015891:Cystatins; D015853:Cysteine Proteinase Inhibitors; D006023:Glycoproteins / Q000276:immunology*; D006801:Humans; D006967:Hypersensitivity / Q000276:immunology*; D007124:Immunoenzyme Techniques; D051379:Mice; D010206:Papain / Q000276:immunology; D011852:Radioallergosorbent Test
chemical_list: D000911:Antibodies, Monoclonal; D039741:Antigens, Dermatophagoides; D015891:Cystatins; D015853:Cysteine Proteinase Inhibitors; D006023:Glycoproteins; D001963:Bromelains; D010206:Papain