RESUMEN
Plaque psoriasis presents with focal skin inflammation, partially maintained by IL-17-mediated interactions between infiltrating epidermal T cells and activated keratinocytes. Here we show that the majority of lesional epidermal CD8 T cells express granzyme A, alone or in combination with IL-17. To assess proinflammatory properties of granzyme A in psoriasis, primary human keratinocytes were stimulated with granzyme A in the presence or absence of IL-17. Out of 33 analysed keratinocyte-derived inflammatory mediators, granzyme A potentiated IL-17-induced secretion of CXCL 1, CXCL 12 and CCL 4. Intriguingly, all three chemokines are implicated in psoriasis pathogenesis and are involved in recruitment of T cells, neutrophils and pDCs into inflamed tissues. Our results indicate that granzyme A produced by lesional CD8 T cells specifically increase the chemokine production from inflamed keratinocytes, thereby amplifying a chemotactic inflammatory loop that sustains psoriasis lesions.
Asunto(s)
Linfocitos T CD8-positivos/enzimología , Granzimas/metabolismo , Interleucina-17/metabolismo , Queratinocitos/metabolismo , Psoriasis/inmunología , Adulto , Anciano , Estudios de Casos y Controles , Femenino , Humanos , Masculino , Persona de Mediana Edad , Psoriasis/enzimologíaRESUMEN
Most psoriasis susceptibility genes were identified in cohorts of mixed clinical phenotypes and the exploration of genes in clinical subtypes is scarce. IL-22 has an established role in host defense and in psoriasis skin pathology, reflecting the delicate balance between control of infection and immunopathology. In a case-control study, we compared the genetic association to IL22 in psoriasis onset in patients between 0-9 (n=207), 10-20 (n=394), and 21-40 (n=468) years with healthy controls (n=1,529). Logistic regression analysis revealed association to regulatory elements in the IL22 promoter confined to onset of psoriasis before puberty (odds ratio=1.45, P<0.0007). The associated variants contain putative binding sites for AhR, a potent inducer of IL-22 expression. In a luciferase assay, transcriptional activity of a high-risk gene variant resulted in 80% higher promoter activity (P=0.012) compared with a low-risk variant. Ex vivo stimulated T cells from peripheral blood were analyzed with flow cytometry. Children with psoriasis carrying a high-risk variant produced 1.7 times more IL-22 compared with low-risk variants (P=0.042). Our combined genetic and functional data support the notion that a genetic IL22 variant that promotes epithelial barrier defense is preferentially enriched in and may precipitate the onset of psoriasis at an early age.
Asunto(s)
Regulación de la Expresión Génica , Interleucinas/genética , Regiones Promotoras Genéticas , Psoriasis/genética , Linfocitos T/inmunología , Linfocitos T/metabolismo , Adolescente , Adulto , Factores de Edad , Sitios de Unión , Estudios de Casos y Controles , Niño , Preescolar , Femenino , Variación Genética , Genotipo , Haplotipos , Humanos , Lactante , Recién Nacido , Masculino , Psoriasis/inmunología , Receptores de Hidrocarburo de Aril/metabolismo , Análisis de Regresión , Análisis de Secuencia de ADN , Adulto Joven , Interleucina-22RESUMEN
BACKGROUND: Mycobacterium ulcerans is the causative agent of necrotizing skin ulcerations in distinctive geographical areas. M. ulcerans produces a macrolide toxin, mycolactone, which has been identified as an important virulence factor in ulcer formation. Mycolactone is cytotoxic to fibroblasts and adipocytes in vitro and has modulating activity on immune cell functions. The effect of mycolactone on keratinocytes has not been reported previously and the mechanism of mycolactone toxicity is presently unknown. Many other macrolide substances have cytotoxic and immunosuppressive activities and mediate some of their effects via production of reactive oxygen species (ROS). We have studied the effect of mycolactone in vitro on human keratinocytes--key cells in wound healing--and tested the hypothesis that the cytotoxic effect of mycolactone is mediated by ROS. METHODOLOGY/PRINCIPAL FINDINGS: The effect of mycolactone on primary skin keratinocyte growth and cell numbers was investigated in serum free growth medium in the presence of different antioxidants. A concentration and time dependent reduction in keratinocyte cell numbers was observed after exposure to mycolactone. Several different antioxidants inhibited this effect partly. The ROS inhibiting substance deferoxamine, which acts via chelation of Fe(2+), completely prevented mycolactone mediated cytotoxicity. CONCLUSIONS/SIGNIFICANCE: This study demonstrates that mycolactone mediated cytotoxicity can be inhibited by deferoxamine, suggesting a role of iron and ROS in mycolactone induced cytotoxicity of keratinocytes. The data provide a basis for the understanding of Buruli ulcer pathology and the development of improved therapies for this disease.
Asunto(s)
Antioxidantes/farmacología , Toxinas Bacterianas/farmacología , Proliferación Celular/efectos de los fármacos , Queratinocitos/efectos de los fármacos , Sal Disódica del Ácido 1,2-Dihidroxibenceno-3,5-Disulfónico/farmacología , Adulto , Toxinas Bacterianas/aislamiento & purificación , Catalasa/farmacología , Línea Celular , Supervivencia Celular/efectos de los fármacos , Cromanos/farmacología , Deferoxamina/farmacología , Relación Dosis-Respuesta a Droga , Humanos , Peróxido de Hidrógeno/farmacología , Queratinocitos/citología , Queratinocitos/metabolismo , Macrólidos , Mycobacterium ulcerans/metabolismo , Oxidantes/farmacología , Especies Reactivas de Oxígeno/metabolismo , Sideróforos/farmacología , Factores de TiempoRESUMEN
Several prostaglandin analogues used for glaucoma treatment have been shown to cause increased iridial pigmentation as side-effect. In the present study we identified the types of prostanoid receptors and cyclooxygenase (COX) enzymes that are expressed in human iridial melanocytes isolated from eyes of different colours. Iris specimens were obtained during trabeculectomy surgery, or from enucleated eyes, and the iridial melanocytes were isolated and cultivated. The transcription of the DP, EP1, EP2, EP3, EP4, FP, IP and TP prostanoid receptor genes as well as the COX-1 and COX-2 enzyme genes was investigated using reverse transcriptase-polymerase chain reaction (RT-PCR). Of the prostanoid receptors the FP receptor gene was found to be most consistently transcribed in the melanocytes isolated from both blue- and hazel-coloured eyes. No RNA of the DP, EP2 and TP receptor genes could be detected, whereas the EP1, EP3, EP4 and IP receptor genes were found to be transcribed in melanocytes from some eyes. The COX-2 gene was found to be transcribed, but the COX-1 gene less consistently. There was no difference in gene transcription pattern between melanocytes originating from eyes treated with latanoprost, and eyes not previously treated with the prostaglandin. These results indicate that the FP prostanoid receptor gene is transcribed in cultivated human iridial melanocytes of both blue and hazel eyes, whereas the other prostanoid receptor genes seem to be transcribed much less frequently, or not at all. Surprisingly, the COX-2 rather than the COX-1 gene, was found to be transcribed in the melanocytes.
Asunto(s)
Color del Ojo , Iris/citología , Isoenzimas/genética , Melanocitos/fisiología , Prostaglandina-Endoperóxido Sintasas/genética , Receptores de Prostaglandina/genética , Transcripción Genética , Anciano , Anciano de 80 o más Años , Células Cultivadas , Ciclooxigenasa 1 , Ciclooxigenasa 2 , Femenino , Humanos , Iris/metabolismo , Latanoprost , Masculino , Melanocitos/citología , Proteínas de la Membrana , Persona de Mediana Edad , Prostaglandinas F Sintéticas/farmacología , Receptores de Prostaglandina/metabolismoRESUMEN
Several prostaglandin analogues used for glaucoma treatment cause increased pigmentation of the iris. The purpose of the present study was investigate whether latanoprost, a PGF(2 alpha) analogue, has any effect on the production of endogenous prostaglandins in iridial melanocytes, which could be important in the mechanism leading to increased pigmentation. Bovine and human iridial melanocytes in culture were used for the experiments. Production of endogenous prostaglandins was measured by enzyme immunoassay, and the melanin content was measured spectrophotometrically. In bovine iridial melanocytes, latanoprost acid caused a significant increase of the PGE(2) production, which could be blocked by indomethacin and NS398, indicating an involvement of cyclo-oxygenase 2. In order to study the selectivity of the phenomenon other endogenous substances/drugs were tested, e.g., acetylcholine, carbachol, noradrenaline, neuropeptide Y, substance P and alpha-MSH, but none was found to have any significant effect. Human iridial melanocytes also responded to latanoprost acid with increased production of PGE(2) and in 1 out of 5 individuals increased melanogenesis coincided with increased PGE(2) production. In bovine iridial melanocytes, latanoprost acid did not stimulate melanogenesis. These results indicate that latanoprost acid cause enhanced formation of endogenous prostaglandins that may have auto- and/or paracrine effects in the melanocytes, possibly associated with melanogenesis.
Asunto(s)
Dinoprostona/metabolismo , Iris/efectos de los fármacos , Melanocitos/efectos de los fármacos , Prostaglandinas F Sintéticas/farmacología , Anciano , Anciano de 80 o más Años , Animales , Bovinos , Células Cultivadas , Femenino , Humanos , Iris/citología , Latanoprost , Masculino , Melanocitos/metabolismo , Persona de Mediana Edad , Epitelio Pigmentado Ocular/citología , Tromboxano B2/metabolismo , TrabeculectomíaRESUMEN
BACKGROUND: A three-fold increased risk for glioma among first degree relatives (FDR) to glioma patients has previously been shown. This study compared familial cases with sporadic controls of glioma to see if phenotypic differences could be detected. Different pathways to tumour growth and progression were investigated including cell cycle regulating genes (p53) and growth factors (epidermal growth factor receptor, EGFR), angiogenesis (vascular endothelial growth factor, VEGF and microvessel density, MVD), pathways of detoxification (glutathione-S-transferase, GST pi) and multidrug resistance (P-glycoprotein, Pgp). MATERIALS AND METHODS: Thirty-seven cases of familial gliomas, identified in a population-based study, were compared to 58 sporadic glioma controls chosen with a negative family history of glioma. The immunohistochemistry was performed with standard procedures using the LABSA kit (Zymed lab). RESULTS: Familial cases had significantly more frequent p53- and Pgp-negative tumours, also when correcting for age, sex and histopathology. However, Pgp was no longer significant after correcting for p53 status indicating a correlation between Pgp and p53. A significant difference between VEGF-negative to VEGF-positive tumours (low- or high-grade) was shown, but it was no longer significant when correcting for p53 status. CONCLUSION: Our study investigated phenotypic differences of familial glioma compared to sporadic control. Our finding of a distinct pattern of increased p53- and Pgp-negativity among cases warrants further investigation.