Localization of tryptase to human cutaneous mast cells and keratinocytes by immunofluorescence and immunoperoxidase cytochemistry with monoclonal antitryptase antibody

Abstract

A monoclonal antibody (H4) against tryptase purified from human pulmonary mast cells was prepared and used as an immunoreactive marker for the cellular localization of tryptase in normal human skin and in lesional skin from subjects with systemic mastocytosis. Mast cells had characteristic metachromatic staining of cytoplasmic granules with Giemsa reagent and were detected in small numbers about superficial vessels in the papillary dermis of nonlesional skin and in large numbers about deep as well as superficial vessels of lesional skin. By both direct immunofluorescence with fluorescein isothiocyanate-H4 and indirect immunoperoxidase cytochemistry with H4, all mast cells were selectively stained. The reactivity was confined to the cytoplasm and was granular in character. In addition, keratinocytes in epidermal tissue and in cell suspensions stained diffusely with H4 antibody. A tryptase-like activity that cleaved tosyl- l-arginine methyl ester (0.003 U/10 6 cells) and was not inhibited by soy bean and lima bean trypsin inhibitors was detected in sonicated suspensions of purified epidermal keratinocytes. Monoclonal antitryptase antibody represents an immunologic probe for the presence of tryptase, a preformed mediator of human mast cells, in tissues and cells.