RESUMEN
The hepatitis C virus (HCV) encodes approximately 10 different structural and non-structural proteins, including the envelope glycoprotein 2 (E2). HCV proteins, especially the envelope proteins, bind to cell receptors and can damage tissues. Endothelial inflammation is the most important determinant of fibrosis progression and, consequently, cirrhosis. The aim of this study was to evaluate and compare the inflammatory response of endothelial cells to two recombinant forms of the HCV E2 protein produced in different expression systems (Escherichia coli and Pichia pastoris). We observed the induction of cell death and the production of nitric oxide, hydrogen peroxide, interleukin-8 and vascular endothelial growth factor A in human umbilical vein endothelial cells (HUVECs) stimulated by the two recombinant E2 proteins. The E2-induced apoptosis of HUVECs was confirmed using the molecular marker PARP. The apoptosis rescue observed when the antioxidant N-acetylcysteine was used suggests that reactive oxygen species are involved in E2-induced apoptosis. We propose that these proteins are involved in the chronic inflammation caused by HCV.
Asunto(s)
Hepacivirus/metabolismo , Células Endoteliales de la Vena Umbilical Humana/inmunología , Células Endoteliales de la Vena Umbilical Humana/patología , Óxido Nítrico/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Proteínas del Envoltorio Viral/metabolismo , Apoptosis/genética , Arginasa/metabolismo , Supervivencia Celular , Escherichia coli/metabolismo , Fibrosis , Expresión Génica/genética , Ingeniería Genética/métodos , Vectores Genéticos/metabolismo , Hepacivirus/inmunología , Antígenos de la Hepatitis C/metabolismo , Humanos , Inflamación/metabolismo , Interleucina-8/metabolismo , Pichia/metabolismo , Plásmidos/metabolismo , Proteínas Recombinantes , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
The hepatitis C virus (HCV) encodes approximately 10 different structural and non-structural proteins, including the envelope glycoprotein 2 (E2). HCV proteins, especially the envelope proteins, bind to cell receptors and can damage tissues. Endothelial inflammation is the most important determinant of fibrosis progression and, consequently, cirrhosis. The aim of this study was to evaluate and compare the inflammatory response of endothelial cells to two recombinant forms of the HCV E2 protein produced in different expression systems (Escherichia coli and Pichia pastoris). We observed the induction of cell death and the production of nitric oxide, hydrogen peroxide, interleukin-8 and vascular endothelial growth factor A in human umbilical vein endothelial cells (HUVECs) stimulated by the two recombinant E2 proteins. The E2-induced apoptosis of HUVECs was confirmed using the molecular marker PARP. The apoptosis rescue observed when the antioxidant N-acetylcysteine was used suggests that reactive oxygen species are involved in E2-induced apoptosis. We propose that these proteins are involved in the chronic inflammation caused by HCV.
Asunto(s)
Humanos , Hepacivirus/metabolismo , Células Endoteliales de la Vena Umbilical Humana/inmunología , Células Endoteliales de la Vena Umbilical Humana/patología , Óxido Nítrico/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , Proteínas del Envoltorio Viral/metabolismo , Apoptosis/genética , Arginasa/metabolismo , Supervivencia Celular , Escherichia coli/metabolismo , Fibrosis , Expresión Génica/genética , Ingeniería Genética/métodos , Vectores Genéticos/metabolismo , Hepacivirus/inmunología , Antígenos de la Hepatitis C/metabolismo , Inflamación/metabolismo , /metabolismo , Pichia/metabolismo , Plásmidos/metabolismo , Proteínas Recombinantes , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
For many fungal diseases, macrophages are the major cell population implicated in host protection, primarily by their ability to eliminate the invading fungal pathogen through phagocytosis. In sporotrichosis, this remains true, because of macrophages' ability to recognize Sporothrix schenckii through specific receptors for some of the fungus' cellular surface constituents. Further confirmation for macrophages' pivotal role in fungal diseases came with the identification of toll-like receptors, and the subsequent numerous associations found between TLR-4 deficiency and host susceptibility to diverse fungal pathogens. Involvement of TLR-4 in immune response against sporotrichosis has been conducted to investigate how TLR-4 signaling could affect inflammatory response development through evaluation of H2O2 production and IL-1ß, IL-6 and TGF-ß release during the course of S. schenckii infection on TLR-4-deficient mice. The results showed that macrophages are largely dependent on TLR-4 for inflammatory activation and that in the absence of TLR-4 signaling, increased TGF-ß release may be one of the contributing factors for the abrogated inflammatory activation of peritoneal exudate cells during mice sporotrichosis.