RESUMEN
The cutaneous T cell-attracting chemokine (CTACK)/CCL27 is indispensable in skin inflammation. CTACK/CCL27 is exclusively produced by epidermal keratinocytes to attract CCR10-expressing T lymphocytes to the skin. We investigated the mechanism of CTACK/CCL27 production from normal human epidermal keratinocytes (NHEKs) by the proinflammatory cytokines TNFα and IFNγ. CTACK/CCL27 production was induced by TNFα via ERK, JNK, p38, and NFκB. The induction of CTACK/CCL27 by TNFα was suppressed by IFNγ via a pathway dependent on JAK, STAT1, and STAT3. Our results also demonstrated that IFNγ and TNFα induced the phosphorylation of EGFR and the following phosphorylation of ERK, which is partly responsible for the suppressive effect of IFNγ on TNFα-induced production of CTACK/CCL27. Peri-lesional skin of psoriasis demonstrates early inflammatory changes as we have previously reported. CTACK/CCL27 expression was diffuse in the peri-lesional epidermis, while it was restricted to basal layer in lesional epidermis, suggesting that CTACK/CCL27 expression was induced in the early stage of psoriatic plaque formation, and IFNγ could participate in the suppression of CTACK/CCL27 expression in the lesional epidermis, reflecting the later stage of psoriatic plaque formation. Our study suggests that CTACK/CCL27 may have a pivotal role in the early stage of psoriasis plaque formation, but should be downregulated in the later stage to induce inflammation characteristic for chronic psoriasis plaques.
Asunto(s)
Quimiocina CCL27/genética , Receptores ErbB/genética , Interferón gamma/genética , Psoriasis/genética , Quimiocina CCL27/biosíntesis , Epidermis/crecimiento & desarrollo , Epidermis/metabolismo , Receptores ErbB/metabolismo , Regulación del Desarrollo de la Expresión Génica , Humanos , Inflamación/genética , Inflamación/patología , Interferón gamma/metabolismo , Queratinocitos/metabolismo , Psoriasis/patología , Transducción de Señal , Piel/metabolismo , Piel/patologíaRESUMEN
Gout occurs in individuals with hyperuricemia when monosodium urate (MSU) crystals precipitate in tissues and induce acute inflammation via phagocytic cells such as monocytes. MSU crystals have been demonstrated in skin diseases such as tophaceous gout or psoriasis; however, the importance of MSU crystals in the skin is totally unknown. In this study, we found that MSU crystals, through P2Y(6) receptors, stimulated normal human keratinocytes (NHK) to produce IL-1α, IL-8/CXCL8, and IL-6. P2Y(6) receptor expression increased in MSU-stimulated NHK. Both P2Y(6)-specific antagonist and P2Y(6) antisense oligonucleotides significantly inhibited the production of IL-1α, IL-8/CXCL8, and IL-6 by NHK. Similarly, the P2Y(6)-specific antagonist completely inhibited the MSU-induced production of IL-1ß by THP-1 cells, a human monocytic cell line. Remarkably, the P2Y(6)-specific antagonist significantly reduced neutrophil influx in both mouse air pouch and peritonitis models. Thus, these results indicate that the P2Y(6) receptor signaling pathway may be a potential therapeutic target for MSU-associated inflammatory diseases, such as tophaceous gout.
Asunto(s)
Antioxidantes/efectos adversos , Queratinocitos/inmunología , Psoriasis/inmunología , Antagonistas del Receptor Purinérgico P2/inmunología , Transducción de Señal/efectos de los fármacos , Ácido Úrico/efectos adversos , Animales , Antioxidantes/farmacología , Línea Celular , Citocinas/inmunología , Citocinas/metabolismo , Modelos Animales de Enfermedad , Gota/inmunología , Gota/metabolismo , Gota/patología , Humanos , Hiperuricemia/inmunología , Hiperuricemia/metabolismo , Hiperuricemia/patología , Inflamación/inducido químicamente , Inflamación/inmunología , Inflamación/metabolismo , Inflamación/patología , Queratinocitos/metabolismo , Queratinocitos/patología , Ratones , Ratones Endogámicos BALB C , Monocitos/inmunología , Monocitos/metabolismo , Monocitos/patología , Infiltración Neutrófila/efectos de los fármacos , Infiltración Neutrófila/inmunología , Peritonitis/inmunología , Peritonitis/metabolismo , Peritonitis/patología , Psoriasis/inducido químicamente , Psoriasis/metabolismo , Psoriasis/patología , Antagonistas del Receptor Purinérgico P2/metabolismo , Receptores Purinérgicos P2 , Transducción de Señal/inmunología , Ácido Úrico/farmacologíaRESUMEN
Lysophosphatidic acid (LPA) is a potent lipid mediator with a wide variety of biological actions mediated through G protein-coupled receptors (LPA(1-6)). LPA(4) has been identified as a G(13) protein-coupled receptor, but its physiological role is unknown. Here we show that a subset of LPA(4)-deficient embryos did not survive gestation and displayed hemorrhages and/or edema in many organs at multiple embryonic stages. The blood vessels of bleeding LPA(4)-deficient embryos were often dilated. The recruitment of mural cells, namely smooth muscle cells and pericytes, was impaired. Consistently, Matrigel plug assays showed decreased mural cell coverage of endothelial cells in the neovessels of LPA(4)-deficient adult mice. In situ hybridization detected Lpa4 mRNA in the endothelium of some vasculatures. Similarly, the lymphatic vessels of edematous embryos were dilated. These results suggest that LPA(4) regulates establishment of the structure and function of blood and lymphatic vessels during mouse embryogenesis. Considering the critical role of autotaxin (an enzyme involved in LPA production) and Gα(13) in vascular development, we suggest that LPA(4) provides a link between these 2 molecules.
Asunto(s)
Vasos Sanguíneos/embriología , Desarrollo Embrionario/fisiología , Vasos Linfáticos/embriología , Receptores Purinérgicos/metabolismo , Animales , Vasos Sanguíneos/metabolismo , Northern Blotting , Southern Blotting , Femenino , Inmunohistoquímica , Hibridación in Situ , Vasos Linfáticos/metabolismo , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
B cells play critical roles in the pathogenesis of lupus. To examine the influence of B cells on disease pathogenesis in a murine lupus model, New Zealand Black and New Zealand White F(1) hybrid (NZB/W) mice were generated that were deficient for CD19 (CD19(-/-) NZB/W mice), a B cell-specific cell surface molecule that is essential for optimal B cell signal transduction. The emergence of anti-nuclear Abs was significantly delayed in CD19(-/-) NZB/W mice compared with wild type NZB/W mice. However, the pathologic manifestations of nephritis appeared significantly earlier, and survival was significantly reduced in CD19(-/-) NZB/W mice compared with wild type mice. These results demonstrate both disease-promoting and protective roles for B cells in lupus pathogenesis. Recent studies have identified a potent regulatory B cell subset (B10 cells) within the rare CD1d(hi)CD5(+) B cell subset of the spleen that regulates acute inflammation and autoimmunity through the production of IL-10. In wild type NZB/W mice, the CD1d(hi)CD5(+)B220(+) B cell subset that includes B10 cells was increased by 2.5-fold during the disease course, whereas CD19(-/-) NZB/W mice lacked this CD1d(hi)CD5(+) regulatory B cell subset. However, the transfer of splenic CD1d(hi)CD5(+) B cells from wild type NZB/W mice into CD19(-/-) NZB/W recipients significantly prolonged their survival. Furthermore, regulatory T cells were significantly decreased in CD19(-/-) NZB/W mice, but the transfer of wild type CD1d(hi)CD5(+) B cells induced T regulatory cell expansion in CD19(-/-) NZB/W mice. These results demonstrate an important protective role for regulatory B10 cells in this systemic autoimmune disease.
Asunto(s)
Anticuerpos Antinucleares/biosíntesis , Antígenos CD19/genética , Subgrupos de Linfocitos B/inmunología , Terapia de Inmunosupresión , Nefritis Lúpica/inmunología , Linfopenia/inmunología , Animales , Anticuerpos Antinucleares/fisiología , Antígenos CD19/metabolismo , Subgrupos de Linfocitos B/metabolismo , Subgrupos de Linfocitos B/patología , Progresión de la Enfermedad , Femenino , Nefritis Lúpica/genética , Nefritis Lúpica/patología , Linfopenia/genética , Linfopenia/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Endogámicos NZB , Ratones NoqueadosRESUMEN
Although contact hypersensitivity (CHS) has been considered a prototype of T cell-mediated immune reactions, recently a significant contribution of regulatory B cell subsets in the suppression of CHS has been demonstrated. CD22, one of the sialic acid-binding immunoglobulin-like lectins, is a B cell-specific molecule that negatively regulates BCR signaling. To clarify the roles of B cells in CHS, CHS in CD22(-/-) mice was investigated. CD22(-/-) mice showed delayed recovery from CHS reactions compared with that of wild-type mice. Transfer of wild-type peritoneal B-1a cells reversed the prolonged CHS reaction seen in CD22(-/-) mice, and this was blocked by the simultaneous injection with IL-10 receptor Ab. Although CD22(-/-) peritoneal B-1a cells were capable of producing IL-10 at wild-type levels, i.p. injection of differentially labeled wild-type/CD22(-/-) B cells demonstrated that a smaller number of CD22(-/-) B cells resided in lymphoid organs 5 d after CHS elicitation, suggesting a defect in survival or retention in activated CD22(-/-) peritoneal B-1 cells. Thus, our study reveals a regulatory role for peritoneal B-1a cells in CHS. Two distinct regulatory B cell subsets cooperatively inhibit CHS responses. Although splenic CD1d(hi)CD5(+) B cells have a crucial role in suppressing the acute exacerbating phase of CHS, peritoneal B-1a cells are likely to suppress the late remission phase as "regulatory B cells." CD22 deficiency results in disturbed CHS remission by impaired retention or survival of peritoneal B-1a cells that migrate into lymphoid organs.
Asunto(s)
Subgrupos de Linfocitos B/inmunología , Inhibición de Contacto/inmunología , Dermatitis por Contacto/inmunología , Peritoneo/citología , Peritoneo/inmunología , Lectina 2 Similar a Ig de Unión al Ácido Siálico/biosíntesis , Lectina 2 Similar a Ig de Unión al Ácido Siálico/genética , Traslado Adoptivo , Animales , Subgrupos de Linfocitos B/citología , Subgrupos de Linfocitos B/patología , Subgrupos de Linfocitos B/trasplante , Movimiento Celular/genética , Movimiento Celular/inmunología , Células Cultivadas , Inhibición de Contacto/genética , Dermatitis por Contacto/metabolismo , Dermatitis por Contacto/patología , Interleucina-10/biosíntesis , Interleucina-10/deficiencia , Interleucina-10/fisiología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Peritoneo/patología , Lectina 2 Similar a Ig de Unión al Ácido Siálico/fisiologíaRESUMEN
Systemic sclerosis (SSc) is a systemic disorder that typically results in fibrosis of the skin and multiple internal organ systems. Although the precise mechanism is unknown, overproduction of extracellular matrix proteins, including collagens and fibronectins, and aberrant immune activation might be involved in the pathogenesis. The soluble cluster of differentiation 21 (sCD21) represents the extracellular portion of the CD21 glycoprotein that is released by shedding from the cell surfaces into plasma. sCD21 binds complement fragments and activates monocytes through binding to membrane CD23. The present study was undertaken to investigate the serum levels of sCD21 in patients with SSc. Serum sCD21 levels were reduced with age both in patients with SSc and normal controls. Serum sCD21 levels in patients with SSc were significantly decreased compared to those in control subjects. When we divided patients with SSc into limited cutaneous SSc (lcSSc) and diffuse cutaneous SSc (dcSSc), patients with lcSSc had lower levels of serum sCD21 than those with dcSSc. Moreover, the prevalence of pulmonary fibrosis in the patients with dcSSc inversely correlated with serum sCD21 levels. Our finding may support the notion that B-cell activation is involved in the mechanism for pulmonary fibrosis and skin sclerosis.
Asunto(s)
Receptores de Complemento 3d/sangre , Esclerodermia Sistémica/sangre , Esclerodermia Sistémica/epidemiología , Adulto , Anciano , Regulación hacia Abajo/fisiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Fibrosis Pulmonar/sangre , Fibrosis Pulmonar/epidemiología , Fibrosis Pulmonar/inmunología , Esclerodermia Sistémica/inmunología , Piel/inmunología , Piel/patología , SolubilidadRESUMEN
Dendritic cells (DCs) are a group of professional antigen-presenting cells, and many genes are known to be associated with their maturation. We compared the transcriptional profiles of immature and mature mouse Langerhans cells using the suppressive, subtractive hybridization method and identified a novel gene of unknown function, termed herein transmembrane protein 123 (Tmem123), of which mRNA expression was enhanced in mature but not in immature Langerhans cells. Its expression was also enhanced in other mature DCs such as bone marrow-derived DCs (BMDCs) and splenic DCs. Interestingly, CD40 expression was up-regulated on mature BMDCs cultured with colchicine concurrently with the enhanced expression of Tmem123 compared with that of fresh BMDCs. Furthermore, the expression of CD40 was enhanced on Tmem123-transfected DC2.4 cells, a mouse BMDC-derived cell line, compared with that on mock-transfected DC2.4 cells. This enhancement of CD40 expression did not occur after deletion of lysosome/endosome targeting YXXÏ motifs (where X is any amino acid and Ï is a bulky hydrophobic amino acid) in the Tmem123 cytoplasmic tail. By stimulation with anti-CD40 monoclonal antibody, these transfectants secreted an increased amount of IL-12/23 p40 compared with mock-transfected DC2.4 cells. Thus, our study demonstrates that Tmem123 may be used as a new maturation marker in DCs and that this molecule may be closely associated with the cell surface expression of CD40.
Asunto(s)
Antígenos CD40/biosíntesis , Células de Langerhans/metabolismo , Proteínas de la Membrana/biosíntesis , ARN Mensajero/biosíntesis , Bazo/metabolismo , Regulación hacia Arriba/fisiología , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Animales , Antígenos CD40/genética , Células COS , Chlorocebus aethiops , Células HeLa , Humanos , Péptidos y Proteínas de Señalización Intracelular , Células de Langerhans/citología , Proteínas de la Membrana/genética , Ratones , Receptores de Superficie Celular , Eliminación de Secuencia , Bazo/citologíaRESUMEN
CD19 is a B-cell transmembrane molecule that is critical for B-cell activation. CD19 serves as a scaffold protein for key signal transduction molecules including Lyn, PI3K, and Vav, by providing docking sites for these molecules via phosphorylation of CD19-Y(513), CD19-Y(482), and CD19-Y(391). We investigated the process of CD19 tyrosine phophorylation during B-cell activation using Ab specific for each of these phosphorylated tyrosines. BCR engagement induced differential tyrosine phosphorylation, as CD19-Y(513) phophorylation occurred first, and CD19-Y(482) phosphorylation was delayed and transient. Different BCR isotypes exhibited distinct patterns of CD19 phosphorylation: IgG-BCR ligation resulted in faster phosphorylation of CD19-Y(513) and more intense phosphorylation of CD19-Y(391) than IgM-BCR ligation. This affected CD19-mediated downstream pathways involving Vav, PI3K, and Akt. Additionally, the phosphorylation profile of CD19 differed distinctly according to its plasma membrane location. CD19 phosphorylated at Y(513) was almost exclusively located within lipid rafts, whereas phosphorylated Y(482) and Y(391) were found both inside and outside of the rafts. Furthermore, the phosphorylation of all three tyrosines was remarkably enhanced and prolonged following the simultaneous stimulation of BCR and CD40. Thus, variations in phosphorylation patterns may contribute to the complexity of CD19-regulated signal transduction.
Asunto(s)
Antígenos CD19/metabolismo , Linfocitos B/metabolismo , Activación de Linfocitos , Procesamiento Proteico-Postraduccional , Animales , Antígenos CD19/química , Antígenos CD19/genética , Linfocitos B/inmunología , Antígenos CD40/inmunología , Línea Celular Tumoral , Cruzamientos Genéticos , Inmunoglobulina G/inmunología , Inmunoglobulina M/inmunología , Linfoma de Células B/patología , Microdominios de Membrana/metabolismo , Ratones , Ratones Endogámicos C57BL , Fosfatidilinositol 3-Quinasas/metabolismo , Fosforilación , Fosfotirosina/análisis , Proteínas Proto-Oncogénicas c-akt/metabolismo , Proteínas Proto-Oncogénicas c-vav/metabolismo , ARN Interferente Pequeño/farmacología , Receptores de Antígenos de Linfocitos B/inmunología , Receptores de Antígenos de Linfocitos B/metabolismo , Transducción de Señal/inmunología , Organismos Libres de Patógenos EspecíficosRESUMEN
OBJECTIVES: To investigate the difference in the dynamics of transforming growth factor ß (TGF-ß) receptors between normal and scleroderma fibroblasts. METHODS: The cell surface expression levels of TGF-ß receptors were determined by biotinylation and immunoprecipitation assay. The dynamics of TGF-ß receptors on the cell surface was determined by the reversible biotinylation assay. The subcellular localisation of TGF-ß receptors was determined by immunoprecipitation using antibodies against clathrin and caveolin. RESULTS: Although the total expression levels of TGF-ß receptors were elevated in scleroderma fibroblasts compared with normal fibroblasts, there was no significant difference in the cell surface expression levels of TGF-ß receptors between these two groups. However, the internalisation rate of TGF-ß receptors was higher in scleroderma fibroblasts compared with normal fibroblasts. Furthermore, caveolin constitutively made a complex with TGF-ß receptors, while the interaction of clathrin with TGF-ß receptors was marginal in scleroderma fibroblasts. CONCLUSIONS: The dynamics of TGF-ß receptors on the cell surface is accelerated in scleroderma fibroblasts. Considering that the activation state of TGF-ß signalling is regulated by a balance between the clathrin-dependent internalisation and the lipid raft-caveolar internalisation, the accumulation of TGF-ß receptors in caveolin-positive vesicles may result in the deceleration of caveolin-dependent internalisation and subsequently lead to the relative acceleration of clathrin-dependent internalisation.
Asunto(s)
Fibroblastos/metabolismo , Receptores de Factores de Crecimiento Transformadores beta/metabolismo , Esclerodermia Sistémica/metabolismo , Biotinilación , Membrana Celular/metabolismo , Células Cultivadas , Humanos , Esclerodermia Sistémica/patología , Piel/metabolismoRESUMEN
Histamine is a biological amine that plays an important role in allergic responses. However, the involvement of histamine signaling in late allergic responses in the skin is poorly understood. Therefore, we attempted to investigate the involvement of histamine signaling in late allergic responses, especially in keratinocytes (KCs). HaCaT KCs and normal human KCs (NHKs) predominantly expressed histamine H1 receptor (H1R) and H2 receptor (H2R). Histamine suppressed tumor necrosis factor α (TNF-α)- and interferon-γ (IFN-γ)-induced production of CC chemokine ligand 17(CCL17), a type 2 T-helper (Th2) chemokine, by HaCaT KCs. It suppressed the phosphorylation of p38 mitogen-activated protein (MAP) kinase, but not that of extracellular signal-regulated kinases (ERKs), and TNF-α- and IFN-γ-induced nuclear factor κB (NFκB) activity. In contrast, histamine enhanced the production of CXC chemokine ligand 10 (CXCL10), a Th1 chemokine, by TNF-α- and IFN-γ-stimulated HaCaT KCs and NHKs. TNF-α- and IFN-γ-induced CXCL10 production was upregulated by suppression of p38 MAP kinase or NF-κB activity, which could explain histamine involvement. We concluded that histamine suppresses CCL17 production by KCs by suppressing p38 MAP kinase and NF-κB activity through H1R and may act as a negative-feedback signal for existing Th2-dominant inflammation by suppressing CCL17 and enhancing CXCL10 production.
Asunto(s)
Quimiocinas/biosíntesis , Histamina/fisiología , Queratinocitos/metabolismo , Receptores Histamínicos H1/fisiología , Células TH1/metabolismo , Células Th2/metabolismo , Secuencia de Bases , Línea Celular , Cartilla de ADN , Humanos , Proteínas Quinasas/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
BACKGROUND: Some chemokines are known to accelerate wound healing. However, there has been no report on the relationship between Thymus and activation-regulated chemokine (TARC)/CC chemokine ligand (CCL) 17 and wound healing. The purpose of this study was to determine whether CCL17 enhances response to cutaneous injury. METHODS: We made a full-thickness dorsal wound in transgenic (Tg) mice, in which CCL17 was overexpressed and in control mice. Wound size was compared over the course of time. We evaluated the effect of CCL17 on fibroblast migration by a Boyden chamber assay and a scratch wound assay. RESULTS: Wound closure in Tg mice was more accelerated than in control mice. CCL17 enhanced nerve growth factor (NGF) production by 2B4, which is mouse T cell hybridoma. Further, in the wound area of Tg mice, the number of CCR4(+) fibroblasts, CCR4(+) lymphocytes and mast cells was increased compared to control mice, as was the number of NGF(+) lymphocytes around the wound area. In vitro assay, CCL17 was shown to enhance the migration of fibroblasts. CONCLUSION: These results suggest that CCL17 accelerates wound healing, mainly by enhancing fibroblast migration, and possibly by increasing NGF(+) lymphocytes and mast cells, which have independently been reported to enhance wound healing.
Asunto(s)
Movimiento Celular/fisiología , Quimiocina CCL17/fisiología , Fibroblastos/citología , Piel/lesiones , Cicatrización de Heridas/fisiología , Animales , Línea Celular , Proliferación Celular , Quimiocina CCL17/genética , Fibroblastos/fisiología , Linfocitos/citología , Linfocitos/fisiología , Mastocitos/citología , Mastocitos/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Modelos Animales , Factor de Crecimiento Nervioso/sangre , Piel/citología , Factores de TiempoRESUMEN
Ectodomain shedding is a proteolytic mechanism by which a transmembrane protein is converted into a secreted form. Pmel17/gp100 is a melanocyte-specific membrane-bound glycoprotein that has amyloid characteristics and forms fibrillar structures in melanosomes after a complex sequence of post-translational processing and trafficking events, including cleavage by a furin-like proprotein convertase (PC). A secreted form of Pmel17 (termed sPmel17) was also thought to be released due to cleavage by a PC. We used multidisciplinary approaches to demonstrate that sPmel17 is released by ectodomain shedding at the juxtamembrane and/or intramembrane motif and to show that this is independent of cleavage by a PC. We further show that sPmel17 consists of 2 fragments linked by disulfide bonds and that the shedding is inhibited at low temperature but not by metalloproteinase inhibitors. Moreover, treatment with a phorbol ester or a calmodulin inhibitor induces Pmel17 shedding. We also refine the reactivity of HMB50 and NKI/beteb, 2 monoclonal antibodies commonly used as melanoma-specific markers. The fact that those antibodies require physically separated domains of Pmel17 sheds interesting light on its 3-dimensional conformation. We conclude that sPmel17 is released by regulated proteolytic ectodomain shedding.-Hoashi, T., Tamaki, K., Hearing, V. J. The secreted form of a melanocyte membrane-bound glycoprotein (Pmel17/gp100) is released by ectodomain shedding.
Asunto(s)
Melanocitos/metabolismo , Glicoproteínas de Membrana/metabolismo , Adenoviridae , Animales , Anticuerpos Monoclonales/metabolismo , Calmodulina/antagonistas & inhibidores , Línea Celular , Línea Celular Tumoral , Glicosilación , Células HeLa , Humanos , Immunoblotting , Inmunoprecipitación , Melanocitos/efectos de los fármacos , Glicoproteínas de Membrana/química , Glicoproteínas de Membrana/genética , Microscopía Fluorescente , Ésteres del Forbol/farmacología , Fosforilcolina/análogos & derivados , Fosforilcolina/farmacología , Ácidos Polimetacrílicos/farmacología , Proproteína Convertasas/metabolismo , Unión Proteica/efectos de los fármacos , Estructura Terciaria de Proteína/genética , Estructura Terciaria de Proteína/fisiología , Conejos , Azida Sódica/farmacología , Sulfonamidas/farmacología , Antígeno gp100 del MelanomaRESUMEN
Melanosomes are organelles specialized for the production of melanin pigment and are specifically produced by melanocytic cells. More than 150 pigmentation-related genes have been identified, including glycoprotein nonmetastatic melanoma protein b (GPNMB). A recent proteomics analysis revealed that GPNMB is localized in melanosomes, and GPNMB is a membrane-bound glycoprotein that shows high homology with a well-known melanosomal structural protein, Pmel17/gp100. In this study, we show that GPNMB is expressed in melanocytes of normal human skin, as well as in human melanoma cells. GPNMB is heavily glycosylated and is enriched in mature (stage III and IV) melanosomes in contrast to MART-1 and Pmel17, which are abundant in early (stage I and II) melanosomes. MART-1 and Pmel17 play critical roles in the maturation of early melanosomes; thus, we speculate that GPNMB might be important in the functions of late melanosomes, possibly their transport and/or transfer to keratinocytes. We also demonstrate that a secreted form of GPNMB is released by ectodomain shedding from the largely Golgi-modified form of GPNMB and that the PKC and Ca(2+) intracellular signaling pathways regulate that shedding. We conclude that GPNMB is a melanosomal protein that is released by proteolytic ectodomain shedding and might be a useful and specific histological marker of melanocytic cells.
Asunto(s)
Biomarcadores de Tumor/metabolismo , Melanocitos/metabolismo , Melanoma/metabolismo , Melanosomas/metabolismo , Glicoproteínas de Membrana/metabolismo , Neoplasias Cutáneas/metabolismo , Línea Celular Tumoral , Medios de Cultivo/metabolismo , Epidermis/metabolismo , Aparato de Golgi/metabolismo , Células HeLa , Humanos , Melanoma/patología , Estadificación de Neoplasias , Estabilidad Proteica , Proteómica , Neoplasias Cutáneas/patología , Antígeno gp100 del MelanomaRESUMEN
BACKGROUND: ß7 Integrin, a cell adhesion molecule, is present in the form of α4ß7 integrin or αEß7 integrin. α4ß7 Integrin is expressed on most leucocytes and is essential for their migration to gut-associated lymphoid tissues by interacting with its primary ligand, mucosal addressin cell adhesion molecule-1, which is preferentially expressed in gut-associated lymphoid tissues. Although the importance of α4ß7 integrin in intestinal inflammation has been established, its role in cutaneous inflammation remains to be elucidated. OBJECTIVE: We sought to investigate the role of ß7 integrin in cutaneous inflammation. METHODS: We used a murine contact hypersensitivity model and examined the role of ß7 integrin by using ß7 integrin-deficient and αE integrin-deficient mice. RESULTS: ß7 Integrin-deficient mice, not αE integrin-deficient mice, are defective in contact hypersensitivity responses. ß7 Integrin deficiency does not affect irritant contact dermatitis. The distribution, migration, and function of antigen presenting cells from ß7 integrin-deficient mice are comparable to those from wild-type mice. Moreover, sensitized ß7 integrin-deficient T cells are able to respond to antigen stimuli in vitro and elicit contact hypersensitivity responses when directly injected into the skin. However, they are defective in reaching the skin under inflammatory conditions, resulting in reduced contact hypersensitivity responses when intravenously injected. Furthermore, intraperitoneal injection of anti-α4ß7 integrin neutralizing antibody elicit impaired contact hypersensitivity responses. CONCLUSION: α4ß7 Integrin contributes to contact hypersensitivity responses by regulating T-cell migration to inflammatory skin.
Asunto(s)
Dermatitis por Contacto/inmunología , Integrinas/metabolismo , Piel/inmunología , Linfocitos T/metabolismo , Traslado Adoptivo , Animales , Anticuerpos Bloqueadores/administración & dosificación , Movimiento Celular/efectos de los fármacos , Movimiento Celular/genética , Células Cultivadas , Dermatitis por Contacto/tratamiento farmacológico , Dermatitis por Contacto/genética , Inmunidad Mucosa/efectos de los fármacos , Integrinas/antagonistas & inhibidores , Integrinas/genética , Integrinas/inmunología , Activación de Linfocitos/efectos de los fármacos , Activación de Linfocitos/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Piel/efectos de los fármacos , Piel/patología , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología , Linfocitos T/patologíaRESUMEN
Given the importance of appropriate diagnosis and appropriate assessment of cutaneous symptoms in treatment of atopic dermatitis, the basics of treatment in this guideline are composed of (1) investigation and countermeasures of causes and exacerbating factors, (2) correction of skin dysfunctions (skin care), and (3) pharmacotherapy, as three mainstays. These are based on the disease concept that atopic dermatitis is a inflammatory cutaneous disease with eczema by atopic diathesis, multi-factorial in onset and aggravation, and accompanied by skin dysfunctions. These three points are equally important and should be appropriately combined in accordance with the symptoms of each patient. In treatment, it is important to transmit the etiological, pathological, physiological, or therapeutic information to the patient to build a favorable partnership with the patient or his/her family so that they may fully understand the treatment. This guideline discusses chiefly the basic therapy in relation to the treatment of this disease. The goal of treatment is to enable patients to lead an uninterrupted social life and to control their cutaneous symptoms so that their quality of life (QOL) may meet a satisfactory level.
Asunto(s)
Dermatitis Atópica/diagnóstico , Dermatitis Atópica/terapia , Dermatitis Atópica/complicaciones , Dermatitis Atópica/fisiopatología , Progresión de la Enfermedad , Humanos , Japón , Derivación y Consulta , Factores de Riesgo , Cuidados de la PielRESUMEN
Sarcoidosis is a chronic multisystem disease of unknown origin, which is characterized by T-helper-1 (Th1)-mediated immune responses. On the other hand, atopic dermatitis (AD) is characterized by Th2-mediated immune responses. We recently experienced an interesting case of AD in which the patient experienced a significant resolution of AD after the onset of sarcoidosis. We herein describe the details of his clinical course and discuss the impact of negative cross-regulation between Th1 and Th2 immune responses.
Asunto(s)
Dermatitis Atópica/fisiopatología , Sarcoidosis/fisiopatología , Células TH1/fisiología , Células Th2/fisiología , Adulto , Biopsia , Dermatitis Atópica/inmunología , Humanos , Masculino , Prednisona/administración & dosificación , Sarcoidosis/inmunología , Tomografía Computarizada por Rayos XRESUMEN
Psoriasis vulgaris is an autoimmune dermatosis with Th17 infiltration. Prolactin (PRL) may participate in the pathogenesis of psoriasis. The chemokine CCL20 recruits Th17 cells, and CCL20 production by epidermal keratinocytes is enhanced in psoriatic lesions. We examined the in vitro effects of PRL on CCL20 production in human keratinocytes. PRL increased basal and IL-17-induced CCL20 secretion, and mRNA expression in keratinocytes. CCL20 production by PRL was suppressed by antisense oligonucleotides against the AP-1 components c-Fos and c-Jun, whereas that by IL-17 was suppressed by antisense NF-kappaB p50 and p65. CCL20 production induced by PRL plus IL-17 was suppressed by antisense c-Fos, c-Jun, p50, and p65. PRL alone increased the transcriptional activity of AP-1, and c-Fos and c-Jun expression; moderately enhanced NF-kappaB activity and IkappaBalpha phosphorylation; and potently increased IL-17-induced NF-kappaB activity. MEK and JNK inhibitors suppressed PRL- or PRL-plus-IL-17-induced CCL20 production and AP-1 activities. MEK inhibitor suppressed PRL-induced c-Fos expression, whereas JNK inhibitor suppressed c-Jun expression. PRL induced ERK and JNK phosphorylation. These results suggest that PRL may enhance basal and IL-17-induced CCL20 production in keratinocytes by AP-1 and NF-kappaB activation, which is partially mediated via MEK/ERK and JNK. PRL may promote Th17 infiltration into psoriatic lesions via CCL20.
Asunto(s)
Quimiocina CCL20/biosíntesis , Queratinocitos/efectos de los fármacos , FN-kappa B/metabolismo , Prolactina/farmacología , Factor de Transcripción AP-1/metabolismo , Células Cultivadas , Quimiocina CCL20/genética , Humanos , Interleucina-17/farmacología , Queratinocitos/inmunología , FN-kappa B/inmunología , Oligonucleótidos Antisentido/genética , Fosforilación , Proteínas Quinasas/metabolismo , Psoriasis/inmunología , Piel/inmunología , Piel/patología , Factor de Transcripción AP-1/inmunologíaRESUMEN
BACKGROUND: Anti-cyclic citrullinated peptide antibodies (anti-CCP) are reported to be found in 5-13% of patients with psoriatic arthritis (PsA). However, whether anti-CCP-positive PsA patients and rheumatoid arthritis (RA) patients have a similar pathophysiological background still remains uncertain. OBJECTIVE: To determine the prevalence of anti-CCP antibodies in patients with PsA and characterize these anti-CCP-positive patients of PsA. METHODS: We measured the serum levels of the anti-CCP antibodies in patients with PsA (n=16), psoriasis (n=15), RA (n=9) and healthy controls (n=11). Serum levels of rheumatoid factor (RF), matrix metalloproteinase-3 (MMP-3), cartilage oligomeric matrix protein (COMP), interleukin (IL)-23p19 and IL-12p40 were also measured in all the samples. RESULTS: Two of the 16 PsA patients (13%) were positive for anti-CCP antibodies with high titers of RF. However, the serum IL-23p19 levels were two orders of magnitude higher in the anti-CCP-positive PsA patients as compared with those in the RA patients and anti-CCP-negative PsA patients. No significant elevation of the serum levels of MMP-3, COMP and IL-12p40 was found in these patients. CONCLUSION: Thirteen percent of the PsA patients were positive for anti-CCP. These patients do not fulfill the American College of Rheumatology (ACR) classification criteria for RA so far. Furthermore, they showed the typical clinical features of PsA rather than those of RA. Although anti-CCP-positive PsA patients may possibly be have a risk of developing RA, we propose that these patients be classified, for the moment, into a independent subtype of PsA, as a different entity from RA.
Asunto(s)
Anticuerpos Antiidiotipos/sangre , Artritis Psoriásica/sangre , Subunidad p19 de la Interleucina-23/sangre , Péptidos Cíclicos/inmunología , Adulto , Artritis Psoriásica/etnología , Artritis Psoriásica/inmunología , Biomarcadores/sangre , Proteína de la Matriz Oligomérica del Cartílago , Estudios de Casos y Controles , Proteínas de la Matriz Extracelular/sangre , Femenino , Glicoproteínas/sangre , Humanos , Subunidad p40 de la Interleucina-12/sangre , Japón , Masculino , Proteínas Matrilinas , Metaloproteinasa 3 de la Matriz/sangre , Persona de Mediana Edad , Factor Reumatoide/sangreRESUMEN
Elevated blood levels of thymus and activation-regulated chemokine (TARC)/CCL17 have been observed in atopic dermatitis (AD) and may serve as a new biomarker for AD. However, the normal levels, especially in children, have not been well determined. We sought to establish an efficient enzyme-linked immunosorbent assay (ELISA) with a wide range of detection that would be suitable for measurement of serum TARC/CCL17 and to determine the normal ranges of this chemokine in different age groups and its diagnostic usefulness for AD. A sensitive specific ELISA for TARC/CCL17, which we previously reported, was modified to accommodate the wide range of TARC/CCL17 values often found in sera. Twenty-seven children with AD under 6 yr of age and 25 age-matched normal non-atopic controls, and 18 patients with AD and 27 controls who were 6 yr and older were enrolled. The severity of AD was evaluated using the SCORAD index. The serum levels of TARC/CCL17 were measured with the ELISA, and the serum levels of IP-10/CXCL10 were also measured. With the novel ELISA system, the assayable range of TARC/CCL17 was 14-8000 pg/ml, and the coefficient of variation at various concentrations ranged from 2.3% to 5.0%. The serum levels of TARC/CCL17 in normal individuals were significantly higher in young children, especially in the age group of 0-1 yr. The cut-off values of TARC/CCL17 for the diagnosis of AD were 1431 pg/ml for 0-1 yr group, 803 pg/ml for 2-5 yr group and 510 pg/ml for the 6 yr and older group, with high sensitivity and specificity of 0.83 and 0.93, 0.83 and 0.92, 0.85 and 0.96, respectively. The magnitude of the decrease in the SCORAD index after treatment with topical steroids correlated significantly with the decrease in serum TARC/CCL17. There was no difference in the serum levels of IP-10/CXCL10 between AD and the controls. The TARC/CCL17:IP-10/CXCL10 ratio tended to be higher in the control children aged 0-1 yr than in those aged 2-5 yr. The serum level of TARC/CCL17 reflects the severity and therapeutic response in AD. The high normal levels in infants should be taken into account when assaying TARC/CCL17.
Asunto(s)
Biomarcadores/sangre , Quimiocina CCL17/sangre , Dermatitis Atópica , Timo/inmunología , Factores de Edad , Quimiocina CXCL10/sangre , Niño , Preescolar , Dermatitis Atópica/sangre , Dermatitis Atópica/diagnóstico , Dermatitis Atópica/inmunología , Dermatitis Atópica/fisiopatología , Ensayo de Inmunoadsorción Enzimática , Humanos , Lactante , Recién Nacido , Índice de Severidad de la Enfermedad , Células TH1/inmunología , Células Th2/inmunologíaRESUMEN
BACKGROUND: Pemphigus is a rare life-threatening intractable autoimmune blistering disease caused by IgG autoantibodies to desmogleins. It has been difficult to conduct a double-blind clinical study for pemphigus partly because, in a placebo group, appropriate treatment often must be provided when the disease flares. OBJECTIVE: A multicenter, randomized, placebo-controlled, double-blind trial was conducted to investigate the therapeutic effect of a single cycle of high-dose intravenous immunoglobulin (400, 200, or 0 mg/kg/d) administered over 5 consecutive days in patients relatively resistant to systemic steroids. METHODS: We evaluated efficacy with time to escape from the protocol as a novel primary end point, and pemphigus activity score, antidesmoglein enzyme-linked immunosorbent assay scores, and safety as secondary end points. RESULTS: We enrolled 61 patients with pemphigus vulgaris or pemphigus foliaceus who did not respond to prednisolone (> or =20 mg/d). Time to escape from the protocol was significantly prolonged in the 400-mg group compared with the placebo group (P < .001), and a dose-response relationship among the 3 treatment groups was observed (P < .001). Disease activity and enzyme-linked immunosorbent assay scores were significantly lower in the 400-mg group than in the other groups (P < .05 on day 43, P < .01 on day 85). There was no significant difference in the safety end point among the 3 treatment groups. LIMITATION: Prednisolone at 20 mg/d or more may not be high enough to define steroid resistance. CONCLUSION: Intravenous immunoglobulin (400 mg/kg/d for 5 d) in a single cycle is an effective and safe treatment for patients with pemphigus who are relatively resistant to systemic steroids. Time to escape from the protocol is a useful indicator for evaluation in randomized, placebo-controlled, double-blind studies of rare and serious diseases.