Keratin intermediate filaments bear antigenic determinants for stratum corneum antibodies.
Stratum corneum (SC) antibodies are directed against antigens in the SC of the epidermis and are known to occur in all normal human sera. They have been shown by indirect immunofluorescence to be frequently associated with upper cytoplasmic (U-Cyt) antibodies. We have recently identified keratin intermediate filaments (KIF) as antigens for U-Cyt antibodies. In this study we investigated whether KIF also bear antigenic sites for SC antibodies. Normal human sera that contained SC and/or U-Cyt antibodies by indirect immunofluorescence were studied. Using immunoblot techniques 3 selected sera were shown to bind to high-molecular-weight (HMW) KIF proteins which had been extracted from 2 different epidermal cell preparations, that is, human callus or epidermis from which the SC had been removed by tape-stripping. The 3 test sera were absorbed on KIF which had been reconstituted in vitro from urea extracts from both epidermal substrates. As shown by indirect immunofluorescence, the SC and U-Cyt antibodies of all 3 sera were absorbed out with KIF from callus and with KIF from epidermis without SC. Immunoblot experiments, which are more sensitive than indirect immunofluorescence, demonstrated the absorption of anti-KIF protein antibodies of the 3 test sera on callus KIF and 2 of the sera on KIF obtained from epidermis without SC. This was shown by the lack of staining of the respective HMW KIF proteins with the postabsorption sera. With the third serum a marked reduction of antibody binding was found after absorption on KIF from epidermis without SC. These data indicate that KIF bear antigenic sites for SC antibodies.
D000042:Absorption; D000906:Antibodies / Q000032:analysis*; D003593:Cytoplasm / Q000276:immunology; D003599:Cytoskeleton / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004817:Epidermis / Q000276:immunology*; D000939:Epitopes / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007633:Keratins / Q000276:immunology*
D000906:Antibodies; D000939:Epitopes; D007633:Keratins