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Generation and Culturing of Primary Human Keratinocytes from Adult Skin
Published on: December 22, 2017
Dynamic expression of pemphigus and desmosomal antigens by cultured keratinocytes
K Iwatsuki1, K Sugaya, M Takigawa
1Department of Dermatology, Fukushima Medical College, Japan.
Abstract:
Dynamic expression of pemphigus antigens by cultured human and mouse keratinocytes was compared with that of desmosome-associated molecules and cellular markers relating to epidermal differentiation. Plakoglobin was detected in localized areas of keratinocyte sheets in low Ca2+ (0.15 mM) KGM medium. In minimum essential medium (MEM) containing 1.8 mM Ca2+, plakoglobin was expressed in the intercellular spaces (ICS) throughout the keratinocyte sheet. Desmoplakin I and II, which were present in the cytoplasm of keratinocytes in the low Ca2+ medium, moved to the cell surface after the medium was changed to MEM. Desmoglein 1 and pemphigus vulgaris (PV) antigens were observed in the ICS of both the monolayers and stratified areas in the MEM. Pemphigus foliaceus (PF) antigens, frequently together with desmoglein 1, involucrin and keratins specific for the upper layer of the epidermis, were expressed by stratified keratinocytes but not the cells in the monolayers. The Western blotting study of the cultured keratinocyte extract showed 160- and 130-kDa bands positive for desmoglein 1 antigens and a 130-kDa band stained with PV sera. These findings suggest that although desmoglein 1 molecules bear PF antigenic sites, their expression pattern by cultured keratinocytes is closely related to that of PV rather than PF antigens. The PF antigenic sites may be formed on desmoglein 1 during epidermal differentiation.
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