Pemphigoid, Pemphigus and Pr Antigens in Adult Human Keratinocytes Grown on Nonviable Substrates

Woodley, David T.; Saurat, J.‐H.; Pruniéras, M; Régnier, M

doi:10.1111/1523-1747.ep12510438

Cited by 27 publications

(3 citation statements)

References 19 publications

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“…Also, laminin ( fig. lb), collagen IV and proteoglycans antigens are revealed by immunofluorescence [14], REp on DED. Adult human skin was obtained after excision in plastic surgery.…”

Section: Reconstructed Epidermismentioning

confidence: 96%

Reconstructed Human Epidermis: A Model to Study in vitro the Barrier Function of the Skin

Régnier

Caron

Reichert

et al. 1992

Skin Pharmacol Physiol

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In previous studies, we have shown that architecture and differentiation of human epidermis reconstructed by culturing dissociated keratinocytes on deepidermized dermis at the air-liquid interface resembles very much those of epidermis in vivo. The various types of epidermal layers are present with the appropriate differentiation markers, such as the bullous pemphigoid antigen, suprabasal keratins, involucrin, membrane-bound transglutaminase and filaggrin, positioned almost like in normal epidermis. At the ultrastructural level, heterogeneous keratohyalin granules and intra- and extracellular membrane-coating granules are observed. After 7 days of culture, a compact stratum corneum covers the reconstructed epidermis. In the present work, the barrier function of the reconstructed epidermis was evaluated by measuring fluxes of ³H-water. Our results show that under the various culture conditions tested, the presence of the reconstructed epidermis reduces dramatically the permeability of the deepidermized dermis although the skin equivalent exhibits a higher permeability than normal skin. Conditions that favor terminal differentiation of the keratinocytes such as maintaining the cultures in delipidized serum improve the barrier function. Reduction of the relative humidity has a similar effect, whereas the age of the culture is of no influence. Experiments are in progress to reconstruct human epidermis with a barrier function as efficient as in normal skin.

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“…Also, laminin ( fig. lb), collagen IV and proteoglycans antigens are revealed by immunofluorescence [14], REp on DED. Adult human skin was obtained after excision in plastic surgery.…”

Section: Reconstructed Epidermismentioning

confidence: 96%

Reconstructed Human Epidermis: A Model to Study in vitro the Barrier Function of the Skin

Régnier

Caron

Reichert

et al. 1992

Skin Pharmacol Physiol

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“…Therefore both PV and PF antigens are expressed in reconstituted skin [47, 55–57]. By culturing keratinocytes air-exposed on a dermal equivalent, it is possible to reconstruct a multilayered epidermis [46, 58].…”

Section: Keratinocyte Culturesmentioning

confidence: 99%

Experimental Human Cell and Tissue Models of Pemphigus

Wier

Pas

Jonkman

2010

Dermatology Research and Practice

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Pemphigus is a chronic mucocutaneous autoimmune bullous disease that is characterized by loss of cell-cell contact in skin and/or mucous membranes. Past research has successfully identified desmosomes as immunological targets and has demonstrated that acantholysis is initiated through direct binding of IgG. The exact mechanisms of acantholysis, however, are still missing. Experimental model systems have contributed considerably to today's knowledge and are still a favourite tool of research. In this paper we will describe to what extent human cell and tissue models represent the in vivo situation, for example, organ cultures of human skin, keratinocyte cultures, and human skin grafted on mice and, furthermore, how suitable they are to study the pathogenesis of pemphigus. Organ cultures closely mimic the architecture of the epidermis but are less suitable to answer posed biochemical questions. Cultured keratinocyte monolayers are convenient in this respect, but their desmosomal make-up in terms of adhesion molecules does not exactly reflect the in vivo situation. Reconstituted skin is a relatively new model that approaches organ culture. In models of human skin grafted on mice, acantholysis can be studied in actual human skin but now with all the advantages of an animal model.

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“…During experiments to show that BP antigen is synthesized by human kératinocytes without the influence of mesenchymal cells [24][25][26], it was discov ered that in cell culture BP antigen was found within the cell as well as being deposited in the extracellular matrix. Fur ther, in cell culture, it appeared that BP anti gen was processed intracellularly and ex pressed first in a perinuclear distribution, then within the Golgi apparatus and finally at the cell plasma membrane in a polar dis tribution [24], For many years there was a problem that haunted investigators studying BP antigen. The problem was that when trypsinized, suspended living kératinocytes were tagged with BP antibodies, less than 5% of the cells would be bound with anti body.…”

Section: Intracellular Bp Antigenmentioning

confidence: 99%