RESUMEN
OBJECTIVES: The aim of this study was to investigate the levels of cytokines in placenta-derived mesenchymal stem cells (MSCs) in normal pregnancies and those with pre-eclampsia. MATERIALS AND METHODS: C5a, CD40 Ligand, G-CSF, GM-CSF, GROalpha, I-309, sICAM-1, IFN-gamma, IL-1alpha, IL-1beta, IL-1ra, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12p70, IL-13, IL-16, IL-17, IL-17E, IL-23, IL-27, IL-32alpha, IP-10, I-TAC, MCP-1, MIF, MIP-1alpha, MIP-1beta, Serpin E1, RANTES, SDF-1, TNFalpha, and sTREM-1 were measured in mesenchymal stem cells using the human cytokine array panel A. The soluble intracellular adhesion molecule-1 (sICAM-1), stromal-derived factor-1 (SDF-1) and monocyte chemotactic protein-1 (MCP-1) were measured by real-time PCR and confirmed by Western blot analysis. RESULTS: MSCs derived from the deciduas of normal pregnancies had significantly elevated levels of sICAM (p=0.000) and SDF-1 (p=0.011), compared to the pregnancies with pre-eclampsia. The level of MCP-1 in the decidua-derived MSCs was not significantly different. No significant difference was observed between normal and pre-eclamptic pregnancies for the amnion-derived MSCs. CONCLUSIONS: The decreased levels of sICAM and SDF-1 found in the decidua-derived MSCs from pre-eclamptic pregnancies might be associated with some of the immunological alterations in pre-eclampsia.
Asunto(s)
Citocinas/metabolismo , Células Madre Mesenquimatosas/inmunología , Placenta/citología , Preeclampsia/inmunología , Embarazo/inmunología , Adulto , Diferenciación Celular/fisiología , Células Cultivadas , Quimiocina CCL2/genética , Quimiocina CCL2/metabolismo , Quimiocina CXCL12/genética , Quimiocina CXCL12/metabolismo , Femenino , Edad Gestacional , Humanos , Inmunofenotipificación , Molécula 1 de Adhesión Intercelular/genética , Molécula 1 de Adhesión Intercelular/metabolismo , Células Madre Mesenquimatosas/citología , Placenta/inmunologíaRESUMEN
Mesenchymal stem cells (MSCs) are capable of self-renewal and differentiation into lineages of mesenchymal tissues that are currently under investigation for a variety of therapeutic applications. The purpose of this study was to compare cytokine gene expression in MSCs from human placenta, cord blood (CB) and bone marrow (BM). The cytokine expression profiles of MSCs from BM, CB and placenta (amnion, decidua) were compared by proteome profiler array analysis. The cytokines that were expressed differently, in each type of MSC, were analyzed by real-time PCR. We evaluated 36 cytokines. Most types of MSCs had a common expression pattern including MIF (GIF, DER6), IL-8 (CXCL8), Serpin E1 (PAI-1), GROalpha(CXCL1), and IL-6. MCP-1, however, was expressed in both the MSCs from the BM and the amnion. sICAM-1 was expressed in both the amnion and decidua MSCs. SDF-1 was expressed only in the BM MSCs. Real-time PCR demonstrated the expression of the cytokines in each of the MSCs. The MSCs from bone marrow, placenta (amnion and decidua) and cord blood expressed the cytokines differently. These results suggest that cytokine induction and signal transduction are different in MSCs from different tissues.
Asunto(s)
Células de la Médula Ósea/citología , Citocinas/metabolismo , Sangre Fetal/citología , Células Madre Mesenquimatosas/metabolismo , Placenta/citología , Citocinas/genética , Femenino , Perfilación de la Expresión Génica , Humanos , Células Madre Mesenquimatosas/citología , Embarazo , Análisis por Matrices de ProteínasRESUMEN
The HOX family of genes plays a fundamental role in the morphogenesis of vertebrate embryonic cells. HOX genes are thought to be important for the regulation of stem cells. We investigated HOX gene expression in mesenchymal stem cells (MSCs) from human placentas. We isolated MSCs from human placentas and confirmed stemness by fluorescence-activated cell sorting (FACS) analysis and differentiation studies. Using reverse transcription PCR, mRNA expression of 39 Class I HOX genes was measured in the MSCs. The expression of HOXB6, C4, C8, C10, D3, D4, and D10 were measured by Western blot analysis. HOXC10 was expressed in 10 of 10 amnion-derived MSCs but in only 2 of 10 decidua-derived MSCs. HOXC4 and D10 were expressed in 100% of both amnion-derived MSCs and deciduas-derived MSCs. HOXD4 was silent in all amnion-derived MSCs and deciduas-derived MSCs (n = 10). HOX gene activation patterns might be a useful indicator for the detection of MSCs of different tissue origins. We demonstrated that HOXC10 is a gene that may discriminate between amnion-derived MSCs and decidua-derived MSCs.
Asunto(s)
Amnios/citología , Biomarcadores/metabolismo , Decidua/citología , Proteínas de Homeodominio/metabolismo , Células Madre Mesenquimatosas/fisiología , Isoformas de Proteínas/metabolismo , Animales , Diferenciación Celular/fisiología , Linaje de la Célula , Separación Celular , Células Cultivadas , Femenino , Perfilación de la Expresión Génica , Regulación de la Expresión Génica , Proteínas de Homeodominio/genética , Humanos , Inmunofenotipificación , Cariotipificación , Células Madre Mesenquimatosas/citología , Embarazo , Isoformas de Proteínas/genéticaRESUMEN
OBJECTIVES: To investigate intralesional cytokine levels in precancerous lesions of the uterine cervix and their relationship with human papillomavirus (HPV) type 16, HPV type 16 E6/E7, and high-risk HPV viral load. METHODS: We performed a prospective study of 67 patients between May 2005 and December 2005. Hybrid Capture II testing was used to identify patients as high-risk HPV DNA-positive or -negative. HPV DNA Chip test was performed for HPV genotyping in all cases found to be HPV DNA-positive. Real-time PCR was used to quantify HPV-16 E6, E7, interleukin-6 (IL-6), interleukin-10 (IL-10), tumor necrosis factor alpha (TNF-alpha), and interferon gamma (IFN-gamma) transcripts. RESULTS: Among high-risk HPV-infected women, intralesional TNF-alpha, IL-6, IL-10, and IFN-gamma levels had no significant differences according to histologic grade. In multivariate logistic regression analysis, TNF-alpha, IL-6, IL-10, and IFN-gamma were not associated with HPV-16. Increased IFN-gamma was significantly associated with HPV-16 E6- and E7-positive (OR 28.197, 95% CI: 2.658-299.110; OR 19.617, 95% CI: 2.135-180.253, respectively), whereas TNF-alpha, IL-6, and IL-10 were not associated with HPV-16 E6 and E7. In multiple regression analysis, elevated IFN-gamma was significantly associated with increased HPV viral load (P=0.039), whereas TNF-alpha, IL-6, and IL-10 were not significantly associated with HPV viral load. CONCLUSIONS: Among HPV-infected women, IFN-gamma is significantly associated with HPV-16 E6, E7, and high-risk HPV viral load in the uterine cervix. Thus, increased intralesional IFN-gamma may be considered to be a prognostic marker for oncogenic potential of high-risk HPV.
Asunto(s)
Citocinas/inmunología , Papillomavirus Humano 16/genética , Proteínas Oncogénicas Virales/biosíntesis , Proteínas Represoras/biosíntesis , Neoplasias del Cuello Uterino/inmunología , Neoplasias del Cuello Uterino/virología , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Genotipo , Humanos , Persona de Mediana Edad , Proteínas Oncogénicas Virales/genética , Proteínas E7 de Papillomavirus , Proteínas Represoras/genética , Carga ViralRESUMEN
Inhibin is a gonadal hormone which is composed of an alpha-subunit and one of two related beta-subunits (betaA, betaB). Inhibin is important for pituitary FSH regulation, normal follicle development and maintenance of the estrous cycle in the female, whereas the role of inhibin in the male is less clear. Thus, we examined the expression of the inhibin-alpha gene in testis during sexual maturation in male mice, to try to gain insight into its functions in the male. Male mice of the ICR strain attained fertility at 6 weeks of age, and histological analysis revealed that a functional testis was formed, with seminiferous tubules which contain mature sperm and with an abundant population of Leydig cells. Parallel with this sexual maturation, inhibin-alpha subunit protein synthesis increased, whereas synthesis of the activin betaA and activin betaB followed with a delayed time course. Inhibin-alpha mRNA also increased during this critical period, and this corresponded to a change in the methylation status of the inhibin-alpha gene. Taken together, our data reveal that activation of inhibin-alpha gene during testis development correlated with the histological maturation of the testis and the acquisition of fertility in male mice.