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Human Epidermal Cells (human + epidermal_cell)
Selected AbstractsCD4 is expressed by epidermal Langerhans' cells predominantly as covalent dimersEXPERIMENTAL DERMATOLOGY, Issue 5 2003G. W. Lynch Abstract:, Langerhans' cells (LC) of skin are CD4 expressing, dendritic, antigen-presenting cells, that are essential for activation of primary immune responses and are productively infected by HIV. We have shown previously that lymphocytes and monocytes express CD4 both as monomers and covalently linked homodimers. In those cells the 55-kDa monomer structure predominates. LC in un-fractionated human epidermal cell (EC) suspension also expresses both forms of CD4, but in EC the dimer form is predominant. Because isolation of LC into single cell suspension by trypsin, as is routinely used for LC isolation, degrades CD4, a systematic study for an alternate procedure for LC isolation was performed. Thus it was found that collagenase blend F treatment can efficiently release LC into suspension, under conditions of only minimal degradation of control soluble recombinant CD4 or CEM-T4 or THP-1 cell CD4, or importantly of LC surface CD4. SDS,PAGE immunoblotting of purified LC extracted from EC by collagenase confirmed CD4 structure as predominantly 110-kDa dimers, with only minimal 55-kDa monomers. The suitability of LC prepared thus for functional studies was demonstrated with binding of functional ligand HIV gp120. It remains to be determined, however, why tissue embedded LC express mainly CD4 dimers, but single-celled blood lymphocytes and monocytes mainly monomers. [source] In vivo UVB irradiation induces clustering of Fas (CD95) on human epidermal cellsEXPERIMENTAL DERMATOLOGY, Issue 6 2003Bo Bang Abstract:,In vitro studies with human cell lines have demonstrated that the death receptor Fas plays a role in ultraviolet (UV)-induced apoptosis. The purpose of the present study was to investigate the relation between Fas expression and apoptosis as well as clustering of Fas in human epidermis after a single dose of UVB irradiation. Normal healthy individuals were irradiated with three minimal erythema doses (MED) of UVB on forearm or buttock skin. Suction blisters from unirradiated and irradiated skin were raised, and Fas, FasL, and apoptosis of epidermal cells were quantified by flow cytometry. Clustering of Fas was demonstrated by confocal laser scanning microscopy on cryostat sections from skin biopsies. Soluble FasL in suction blister fluid was quantified by ELISA. Flow cytometric analysis demonstrated increased expression intensity of Fas after irradiation, with 1.6-, 2.2- and 2.7-fold increased median expression at 24, 48 and 72 h after irradiation, respectively (n = 4). Apoptosis was demonstrated by the TUNEL reaction, and the maximum of apoptotic cells was detected at 48 h after irradiation. Double-staining for Fas and TUNEL showed that apoptosis was restricted to the Fas-positive epidermal subpopulation, but there was no correlation between the intensities of Fas expression and TUNEL reaction. Median expression intensity of FasL-positive cells transiently decreased to 0.9- and 0.8-fold of the preirradiation respective level after 24 h and 48 h, respectively, and returned to the respective preirradiation level at 72 h after irradiation (n = 4). Concentrations of soluble FasL in suction blister fluid from UVB-irradiated skin did not differ from those in unirradiated skin (n = 5). Confocal laser scanning microscopy showed a rapid clustering of Fas within 30 min after irradiation. A simultaneous clustering of the adapter signalling protein FADD suggested that Fas clustering has a functional significance. Our results are in accordance with previous findings from in vitro studies, and suggest that Fas is activated in vivo in human epidermis after UVB exposure. [source] AKT and MAPK signaling in KGF-treated and UVB-exposed human epidermal cellsJOURNAL OF CELLULAR PHYSIOLOGY, Issue 3 2007Lavinia Vittoria Lotti Regulation of proliferation and differentiation in keratinocyte is a complex and dynamic process that involves activation of multiple signaling pathways triggered by different growth factors. Keratinocyte growth factor (KGF) is not only a potent mitogen, but differently from other growth factors, is a potent inducer of differentiation. The MAP kinase and AKT pathways are involved in proliferation and differentiation of many cell types including keratinocytes. We investigated here the role of KGF in modulating AKT and MAPK activity during differentiation of human keratinocytes. Our results show that the mechanisms of action of KGF are dose-dependent and that a sustained activation of the MAPK signaling cascade causes a negative regulation of AKT. We also demostrated increasing expression of KGFR substrates, such as PAK4 during keratinocyte differentiation parallel to the receptor upregulation. J. Cell. Physiol. 212:633,642, 2007. © 2007 Wiley-Liss, Inc. [source] Expression of haptoglobin in human keratinocytes and Langerhans cellsBRITISH JOURNAL OF DERMATOLOGY, Issue 5 2005H. Wang Summary Background, Epidermal Langerhans cells (LCs) play an important role in cutaneous immunological reactions. Freshly obtained or intraepidermal LCs are incapable of activating autologous naive T cells. However, when they are cultured for 2,3 days, LCs are able to activate autologous T cells. It has been proposed that haptoglobin (Hp) is the inhibitor that prevents LC functional transformation in the skin. Abundant Hp has been found in the cytoplasm of epidermal LCs. However, the source of Hp in LCs has not been addressed. Objectives, To determine the expression of Hp in epidermal cells, and to provide evidence that there is a functional relationship between LCs and keratinocytes (KCs) through Hp. Methods, Normal human epidermal cells and HaCaT cells were used for detection of Hp mRNA by in situ hybridization and reverse transcription,polymerase chain reaction, and Hp protein by immunohistochemical staining, immunofluorescence counterstaining and Western blotting. Results, Hp mRNA was expressed in normal human KCs and HaCaT cells, but not in normal human epidermal LCs. Hp protein was detected by immunohistochemical staining and immunofluorescence counterstaining in CD1a+ epidermal dendritic cells (LCs), but not in KCs. Hp protein was weakly expressed by HaCaT cells. Conclusions, Hp mRNA is present in normal human KCs and HaCaT cells, suggesting that they have the potential to synthesize Hp protein. Normal human epidermal LCs are unable to synthesize Hp protein by themselves, although they have abundant Hp protein in their cytoplasm. It is likely that LCs acquire Hp through an exogenous pathway. [source] | ||||||||||