Immunohistochemical study of beta-adrenergic receptors in the psoriatic epidermis using an anti-alprenolol anti-idiotypic antibody

Abstract

In order to study the beta-adrenergic receptor-cyclic AMP system in the human skin, we have developed a monoclonal antibody against the receptor binding site in two fusion steps. We produced an anti-alprenolol monoclonal antibody using a hybridoma technique after the immunization of a mouse with alprenolol-BSA conjugate. This antibody was then used to immunize mice, whose splenocytes were used for the second fusion. Colonies were screened by enzyme linked immunosorbent assay (ELISA) using turkey erythrocyte membrane as an antigen. One of the positive colonies was grown in mice to obtain ascites fluids. This antibody showed the ability to compete with [125I]-iodocyanopindolol for the binding to beta-adrenergic receptors of turkey erythrocytes. The binding of the antibody to the turkey erythrocytes was prevented by the existence of the anti-alprenolol antibody and also L-alprenolol. This antibody showed a prominent inhibition of the adrenaline-stimulated adenylate cyclase activity of the turkey erythrocyte membrane. The data indicate that the anti-idiotypic monoclonal antibody against alprenolol binds to the beta-adrenergic receptors and acts like an antagonist. We further studied the localization of the beta-adrenergic receptors with this antibody in the normal human skin as well as in the psoriatic skin using immunohistochemical technique. A prominent decrease of the beta-receptors was observed in the psoriatic epidermis.