Binding specificity of guinea pig anti-alpha-keratin polypeptide sera on human keratinocytes: comparison of their receptors with those of human epidermal cytoplasmic antibodies.
Experimental anti-keratin polypeptide sera (KPS) which were prepared by immunizing guinea pigs with the P1 polypeptide (molecular weight: 67 0000 dalton) of alpha-keratin of normal human stratum corneum, were shown to react in immunofluorescence only to cell cytoplasmic antigen of the upper layers of the epidermis. No staining was detected in the basal layer. An immunolabeling performed on free epidermal cells obtained after trypsinization demonstrated by electron microscopy that receptors for KPS were tonofilaments. Minor proportions of negative cells containing tonofilaments and numerous melanosomes detected might correspond to the basal cells. An IF pattern similar to that seen with KPS was observed with some human epidermal cytoplasmic sera (ECS). However, reciprocal blocking tests performed on both rabbit lip and normal human skin with the different sera showed no inhibition. No cross-reaction was detected by immunodiffusion test between whole purified alpha-keratin and human ECS. In electron microscopy, the receptors for ECS appeared not to be keratin-like, but were located on a granular part of the peripheral keratinocyte cytoplasm. These findings confirmed two steps (basal and malphighian layers) in epidermal differentiation defined by antigenic markers and especially by the alpha-keratin component of MW 67 000 d present in normal stratum corneum.
D000818:Animals; D000906:Antibodies; D001666:Binding Sites, Antibody; D003593:Cytoplasm / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D004817:Epidermis / Q000276:immunology*; D000939:Epitopes; D005260:Female; D005455:Fluorescent Antibody Technique; D006168:Guinea Pigs; D006801:Humans; D005779:Immunodiffusion; D007124:Immunoenzyme Techniques; D007633:Keratins / Q000276:immunology* / Q000378:metabolism; D008854:Microscopy, Electron; D010455:Peptides / Q000276:immunology*; D011817:Rabbits; D011971:Receptors, Immunologic / Q000276:immunology*; D014357:Trypsin / Q000494:pharmacology
D000906:Antibodies; D000939:Epitopes; D010455:Peptides; D011971:Receptors, Immunologic; D007633:Keratins; D014357:Trypsin