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  • 1
    Electronic Resource
    Electronic Resource
    Expression of Annexin I in freshly isolated human epidermal cells and in cultured keratinocytes (1994)
    Springer
    Archives of dermatological research 286 (1994), S. 268-272 
    Details
    ISSN: 1432-069X
    Keywords: Annexin I ; Human epidermal cells ; Keratinocytes ; Langherans cells ; Inflammation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Annexin I belongs to a newly characterized family of intracellular proteins involved in the regulation of the production of inflammatory lipid mediators such as prostaglandins and leucotrienes. Annexin I (named p35, lipocortin I or calpactin II) was initially described as a protein inducible by glucocorticoids. In the skin, the role of annexins has still not been elucidated. In the study reported here we investigated the expression of annexin I both in freshly isolated epidermal cells and in cultured keratinocytes using immunofluorescence, FACS analysis and immunoblotting techniques. Using epidermal cells freshly isolated from normal skin, annexin I was detected by double immunostaining mainly in basal and suprabasal keratinocytes. Langerhans cells isolated from Ficoll gradient were faintly stained compared with keratinocytes. Annexin I was also highly expressed in keratinocytes maintained in culture in a serum-free medium without hydrocortisone. By confocal microscopy, annexin I was shown to be mainly localized in the cytoplasm of the cells. The protein was characterized by Western blot and immunoprecipitation as a 35-kDa protein in freshly isolated epidermal cells and cultured keratinocytes. Results from in vivo studies confirmed the presence of annexin I in the basal and suprabasal layers of normal human skin with modified reactivity patterns in hyperproliferative lesions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00387599
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  • 2
    Electronic Resource
    Electronic Resource
    Glucocorticoid receptor localization in human epidermal cells (1996)
    Springer
    Archives of dermatological research 288 (1996), S. 140-146 
    Details
    ISSN: 1432-069X
    Keywords: Key words Glucocorticoid receptors ; Human epidermal ; cells ; Keratinocytes ; Langerhans cells ; Melanocytes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Glucocorticoids, which are widely used in therapy, exert their immunosuppressive actions through specific receptors. These receptors have been characterized in cultured human skin fibroblasts and keratinocytes, but their localization in vitro and in vivo has not been established. To determine the tissue and cellular distribution of glucocorticoid receptors (GR), two specific polyclonal rabbit anti-human GR antibodies were used to detect these receptors in skin biopsy specimens, in freshly isolated and cultured human epidermal cells and in keratinocyte cell lines. Immunoreactive GR were only faintly detected in normal and abnormal differentiated cells and as well as those in the stratum granulosum and corneocytes. These immunolocalization studies were confirmed by fluorescence cell sorter analysis of isolated basal and suprabasal keratinocytes. Immunoreactive GR were highly expressed in normal cultured human keratinocytes, Langerhans cells and several cell lines whereas they were less expressed in melanocytes. Based upon these results the main targets of glucocorticoids in the epidermis appear to be basal and Langerhans cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004030050036
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  • 3
    Electronic Resource
    Electronic Resource
    Growth and differentiation of human keratinocytes on extracellular matrix (1987)
    Springer
    Archives of dermatological research 279 (1987), S. 241-246 
    Details
    ISSN: 1432-069X
    Keywords: Cultures ; Epidermal cells ; Extracellular matrices ; Keratinocytes ; Pemphigus ; Pemphigoid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Extracellular matrices (ECM) have been reported to enhance epithelial cell attachment and proliferation as well as to induce differentiation in vitro. Since ECM components are physiological constituents of the dermoepidermal basement membrane, we studied the growth and differentiation of human keratinocytes on ECM in order to determine the benefits of culturing epidermal epithelial cells (keratinocytes) on reconstituted basement membranes. Dissaggregated epidermal cells were grown in primary and subcultures in liquid medium; the attachment of the cells was greatly enhanced by ECM and noted within the first few hours after seeding; cells formed small islets that reached confluence within 2–12 days depending upon the plating density and the type of culture (primary or passages). Histological and ultrastructural crosssections of the cultures clearly indicated that a multilayered epithelium can be obtained including a basal cell layer, several intermediate cell layers with cytoplasmic organelles, intermediate size filaments, desmosomes, and keratohyaline granules, and an upper layer of anucleated cells. Using immunofluorescence, both pemphigus and pemphigoid (basal membrane zone) antigens were expressed. The keratin pattern noted indicated that these epithelia differentiate and keratinize but do not express a complete program of keratinization, a finding usually noted when cells are grown submersed. These data show that ECM favor epidermal cell proliferation and differentiation and suggest that they may be used to obtain large amounts of epidermal equivalent suitable for grafting and/or in vitro studies.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00417322
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  • 4
    Electronic Resource
    Electronic Resource
    Effect of various metals on intercellular adhesion molecule-1 expression and tumour necrosis factor alpha production by normal human keratinocytes (1994)
    Springer
    Archives of dermatological research 286 (1994), S. 466-470 
    Details
    ISSN: 1432-069X
    Keywords: Keratinocytes ; Metals ; ICAM-1 ; TNF-α ; Hsp72
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Nickel, cobalt and chromium are metals very often implicated in allergic contact dermatitis. In vivo, keratinocytes, which are the first target cells, can be directly activated to participate in the local reaction, especially through the expression of the membrane antigen ICAM-1, a ligand of the leucocyte antigen LFA-1, and the production of cytokines. Our aim was to assess the effects of sensitizing metal haptens (nickel, cobalt and chromium) compared with the toxic metal cadmium on the induction of ICAM-1 and the production of TNFα by epidermal cells. For this purpose, normal human keratinocytes obtained during plastic skin surgery were cultured in low-calcium defined medium (MCDB153) and the metals were used in non-toxic concentrations. Using FACS analysis, ICAM-1 expression was found to be induced only by nickel. This stimulation appeared as early as 24 h after stimulation. All the metals induced a low expression of TNFα detectable by immunocytochemistry correlating with the induction of the nuclear stress protein Hsp72 which is closely linked genetically with the TNFα locus. However, only Ni2+, Co2+ and Cr2+ induced a significant release of TNFα detectable by ELISA after 48 h stimulation. This secretion was lower than that observed with known stimulants such as lipopolysaccharide. These results indicate that the metals studied are able to induce an aggressive cellular effect, and that nickel, by its ICAM-1 induction, may play a major role in the keratinocyte activation state during allergic contact dermatitis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00371573
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  • 5
    Electronic Resource
    Electronic Resource
    Effects of thermal shocks on interleukin-1 levels and heat shock protein 72 (HSP72) expression in normal human keratinocytes (1992)
    Springer
    Archives of dermatological research 284 (1992), S. 414-417 
    Details
    ISSN: 1432-069X
    Keywords: Thermal shocks ; Keratinocytes ; Interleukin ; HSP72
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Interleukin-1 expression is reported to be modified under a number of cell conditions including physiological stress, injury and activation. We report the effects of the physiological stresses cold and heat shock on IL-1 levels in keratinocytes. Having observed that normal human skin obtained from plastic surgery, usually stored at 4
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00372072
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  • 6
    Electronic Resource
    Electronic Resource
    Substance P and keratinocyte activation markers: an in vitro approach (1996)
    Springer
    Archives of dermatological research 288 (1996), S. 85-90 
    Details
    ISSN: 1432-069X
    Keywords: Key words Substance P ; Keratinocytes ; ICAM-1 ; Cytokines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Substance P (SP) released by cutaneous C fibres is involved in the physiopathology of cutaneous lesions. As normal human keratinocytes have been reported to express SP receptors, we studied the effects of SP on keratinocyte activation markers such as ICAM-1 induction and cytokine production. Human keratinocytes derived from skin obtained during plastic surgery were cultured in defined medium (MCDB 153) and were stimulated by SP. Flow cytometry analysis showed that SP (10 –7 and 10 –5 M ) as well as the specific NK1 agonist Sar 9 Met(O 2 ) 11 SP (Sar Met) induced a slight but significant expression of ICAM-1 at the cell surface during treatment periods of 24 h and 48 h. SP (10 –5 M ) also induced a significant but transient increase in the production of IL-1α, IL-1β, IL-1 receptor antagonist and IL-8 which was detectable by ELISA techniques 6 h after stimulation. This elevation returned to constitutive levels 24 or 48 h postinduction. TNFα secretion was detected in stimulated cells only after 48 h. These results suggest that SP can activate keratinocytes and support its role in the local inflammatory reaction.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004030050028
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  • 7
    Electronic Resource
    Electronic Resource
    Expression of vascular endothelial growth factor in normal epidermis, epithelial tumors and cultured keratinocytes (1997)
    Springer
    Archives of dermatological research 289 (1997), S. 158-163 
    Details
    ISSN: 1432-069X
    Keywords: Key words Angiogenesis ; Skin tumors ; Keratinocytes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Vascular endothelial growth factor (VEGF), an endothelium-specific growth factor and microvessel hypermeability factor, is expressed and secreted by several kinds of cells and is implicated in angiogenesis of tumors. The present study was performed to determine the relationship between the expression of VEGF in normal skin, benign and malignant epithelial lesions and cultured keratinocytes and the proliferative activity and degree of differentiation of keratinocytes. Skin lesions were studied immunohistochemically by staining with two anti-VEGF antibodies and secretion and production of VEGF by keratinocyte cultures were evaluated using an enzyme-linked immunosorbent assay. Low to moderate VEGF expression was observed in normal epidermis. In epithelial tumors, different reactivity patterns were observed and different areas of the same tumor expressed different amounts of VEGF. A more prominent labelling occurred in proliferative layers and/or more differentiated cells of virus-induced lesions, squamous cell carcinomas and Bowen’s disease, whereas basal cell carcinomas always stained weakly for VEGF. In cultured keratinocytes, the amount of cell-associated and secreted VEGF increased with time, and the constitutively produced VEGF was mostly released extracellularly. High calcium concentrations upregulated the intracellular content of VEGF but downregulated its release. Taken together, these results showed a modulated expression and release of VEGF in relation to the stage of cell differentiation and in rapidly growing or activated keratinocytes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s004030050172
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