Mapping of Lol p I allergenic epitopes by using murine monoclonal antibodies.
Murine monoclonal antibodies (MAbs) against three non-overlapping epitopes of Lol p I allergen were previously produced and subsequently used for purification of the allergen. In the present study, these MAbs were further characterized, and the biological activity of the purified allergen assessed. The three MAbs were of the IgG isotype and carried a kappa light chain. Their affinity constants were in the range of 7.4-15.1 x 10(-9) mol/l. Purified Lol p I kept its biological activity, as shown by its ability to induce histamine release by basophils of Lol p I-sensitive patients. The profiles of histamine release induced by either Lol p I or crude Lolium perenne extracts were comparable. This observation suggests that human IgE bound to basophils are polyspecific which has been confirmed by immunoblot and inhibition assay. Our data indicated also that Lol p I possesses a major allergenic epitope recognized by all human serum IgE tested. This epitope seems to be partially shared by those recognized by the three MAbs. Finally, preincubation of Lol p I with either one of the Mabs did not affect significantly the basophil-histamine release induced by the purified allergen. This suggests that Lol p I possesses allergenic sites other than the one shared by MAbs and IgE Abs.
Mourad
W
W
Unité de Recherche Inflammation et Immunologie-Rhumatologie, Ste-Foy, Canada.
Bernier
D
D
Jobin
M
M
Hébert
J
J
D000485:Allergens; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D052179:Antigens, Plant; D001667:Binding, Competitive / Q000502:physiology; D000939:Epitopes / Q000032:analysis*; D006632:Histamine / Q000378:metabolism; D006801:Humans; D006969:Hypersensitivity, Immediate / Q000276:immunology; D007163:Immunosorbent Techniques; D007962:Leukocytes / Q000378:metabolism; D051379:Mice; D010940:Plant Proteins; D011058:Pollen / Q000032:analysis / Q000276:immunology*; D012434:Secale / Q000276:immunology
D000485:Allergens; D000911:Antibodies, Monoclonal; D052179:Antigens, Plant; D000939:Epitopes; C042814:Lol p I protein, Lolium perenne; D010940:Plant Proteins; D006632:Histamine