RESUMEN
Receptors of the integrin family are largely confined to the basal layer of keratinocytes, both in human epidermis and in stratified cultures of human keratinocytes. However, suprabasal integrin expression is observed during epidermal wound healing and in psoriatic lesions. We have investigated potential stimuli of suprabasal expression. Addition of transforming growth factor-beta (TGF-beta), interferon-gamma (IFN-gamma), or tumor necrosis factor-alpha (TNF-alpha) to keratinocytes cultured with a 3T3 feeder layer did not induce suprabasal expression. The cytokines caused small changes in the levels of alpha 2 beta 1 or alpha 3 beta 1 on the surface of basal keratinocytes but had no significant effect on the proportion of cells adhering to fibronectin, type IV collagen, and laminin, and did not cause changes in the mobility of integrin subunits on polyacrylamide gels. Injection of TNF-alpha or IFN-gamma intradermally into healthy human volunteers induced an inflammatory response but did not induce suprabasal integrin expression. However, we did observe transient suprabasal integrin expression when keratinocytes were grown on a dermal equivalent consisting of fibroblasts in a collagen gel. One week after raising the cultures to the air-liquid interface, beta 1 integrins were found in all the viable cell layers, with suprabasal cells co-expressing integrins and involucrin; 1 week later integrins were confined to the basal layer. Addition of TGF-beta, IFN-gamma, or TNF-alpha to the dermal equivalents neither induced nor inhibited suprabasal integrin expression. We conclude that suprabasal integrin expression is not induced by the inflammatory cytokines tested, and instead may reflect the proliferation/differentiation status of the epidermis.
Asunto(s)
Integrinas/fisiología , Interferón gamma/farmacología , Queratinocitos/química , Factor de Crecimiento Transformador beta/farmacología , Factor de Necrosis Tumoral alfa/farmacología , Células Cultivadas , Citometría de Flujo , Humanos , Recién Nacido , MasculinoRESUMEN
We have examined integrin expression during the remodeling of the epidermis that takes place during wound healing, using a suction blister model in which the epidermis is detached from the dermis, leaving the basement membrane intact. By immunofluorescence microscopy, we found that the same integrin subunits were expressed during wound healing as in normal epidermis with very little change in the relative intensity or distribution of staining at the leading edge of the migrating epidermis. However, at the time of wound closure, when the epidermis is still hyperproliferative, alpha 2, alpha 3, alpha 6, and beta 1 were no longer confined to the basal layer, as in normal epidermis, but were also found in all the living suprabasal cell layers, coexpressed with the terminal differentiation markers involucrin, keratin 10, and keratin 16. Strong suprabasal staining for alpha v was also found in one specimen. beta 4, which normally forms a heterodimer with alpha 6, and alpha 5 remained predominantly basal. Three of the integrin ligands, fibronectin, type IV collagen, and laminin, remained largely confined to the basement membrane zone and dermis. By 14 d after wounding, the integrins were once more restricted to the basal layer. Suprabasal integrin expression was also observed in involved psoriatic lesions. Thus, in two situations in which the epidermis is hyperproliferative, there is a failure to downregulate integrin expression on initiation of terminal differentiation. The functional consequences of this aberrant integrin expression remain to be explored.
Asunto(s)
Epidermis/metabolismo , Integrinas/metabolismo , Psoriasis/metabolismo , Cicatrización de Heridas/fisiología , Vesícula/metabolismo , Diferenciación Celular , Movimiento Celular , Epidermis/patología , Proteínas de la Matriz Extracelular/metabolismo , Humanos , Queratinocitos/citología , Psoriasis/patología , Valores de ReferenciaRESUMEN
In order to investigate the role of extracellular matrix receptors of the integrin family in establishing the spatial organization of epidermal kerotinocytes, we used immunofluorescence microscopy to examine the expression of a range of integrin subunits during development of human palm and sole skin. All of the integrins expressed during development were also present in mature epidermis and were largely confined to the basal layer of keratinocytes in a pericellular distribution. The alpha 3 and beta 1 subunits were expressed prior to the initiation of stratification and did not change in abundance or distribution during subsequent development. alpha 4 and beta 3 were not detected at any time in the epidermis. Every other subunit examined showed spatial or temporal changes in expression. Staining for alpha 1 was strong before stratification and until mid-development, but was greatly decreased in neonatal epidermis. alpha 2 was first detected in small patches of basal cells prior to stratification, and thereafter was found in the entire basal layer, with greater staining in developing sweat glands. alpha 5 was not expressed until mid-development, and then primarily in developing sweat glands, with faint expression in neonatal epidermis. alpha v was detected following stratification, in developing sweat glands, and occasionally in neonatal epidermis. alpha 6 and beta 4 were peribasally expressed before stratification, but thereafter became concentrated at the basal cell surface in contact with the basement membrane, co-localizing with hemidesmosomes as determined by staining with bullous pemphigoid antiserum. We also examined the distribution of three known ligands for keratinocyte integrins: laminin and collagen type IV were present in the basement membrane zone at all stages of development, whereas fibronectin was only evident there until about 13 weeks estimated gestational age. Finally, we found that the changes in integrin expression that occur on initiation of stratification in vivo could be reproduced in organ cultures of developing skin; such cultures therefore provided a useful experimental model for further studies of the role of integrins in epidermal stratification.
Asunto(s)
Desarrollo Embrionario y Fetal/fisiología , Matriz Extracelular/fisiología , Integrinas/genética , Piel/embriología , Pie/embriología , Expresión Génica , Edad Gestacional , Mano/embriología , Humanos , Integrinas/análisis , Microscopía Electrónica , Microscopía Fluorescente , Técnicas de Cultivo de Órganos , Piel/química , Fenómenos Fisiológicos de la PielRESUMEN
Embryonic avian tissue is resistant to the transforming potential of Rous sarcoma virus (RSV) in ovo. Analysis of the pattern of host-viral interactions in the first semester of chick development has demonstrated that RSV is first expressed in a limited population of muscle precursor cells and proceeds to spread throughout the developing dorsal and ventral limb musculature. The number of non-muscle cells participating in the infection is initially low but gradually increases as development continues. The data show that RSV infection in ovo is both compatible with the process of differentiation and the maintenance of the differentiated state of the limb. The kinetics of viral spreading and competence for transformation are developmentally regulated in the embryo. The contrasting properties of embryonic cells in ovo as compared with those of the adult provide an opportunity for evaluating host related regulatory factors that are of significance to the expression of viral transforming function.
Asunto(s)
Virus del Sarcoma Aviar/genética , Transformación Celular Neoplásica , Músculos/embriología , Proteínas de los Retroviridae/análisis , Animales , Virus del Sarcoma Aviar/aislamiento & purificación , Células Cultivadas , Embrión de Pollo , Técnica del Anticuerpo Fluorescente , Productos del Gen gag , Músculos/microbiología , Miosinas/análisisRESUMEN
Tumors induced in chickens by Rous sarcoma virus remain localized at the site of injection even though the animals become viremic. Tumors have now been shown to be inducible at other sites if a wound is inflicted or if the tissue is injured by administration of tumor promoters. These findings indicate that local wounding plays a role in the spread of tumorigenicity of Rous sarcoma virus.