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Inflammatory response of endothelial cells to hepatitis C virus recombinant envelope glycoprotein 2 protein exposure.

Urbaczek AC, Ribeiro LC, Ximenes VF, Afonso A, Nogueira CT, Generoso WC, Alberice JV, Rudnicki M, Ferrer R, Fonseca LM, Costa PI - Mem. Inst. Oswaldo Cruz (2014)

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Imunologia Clínica, Departamento de Análises Clínicas, Escola de Ciências Farmacêuticas, Bauru, SP, Brasil.

ABSTRACT
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.

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: interleukin-8 (IL-8) production by envelope glycoprotein 2(E2)-stimulated human umbilical vein endothelial cells. Results presented asmean and standard deviation. The experiments were performed in triplicate. c-:cells and medium and phosphate-buffered saline (pH 7.2). LPS:lipopolysaccharide (1.0 µg/mL); Sup: culture supernatant Escherichiacoli BL21; TNF-α: tumour necrosis factor alpha (10 ng/mL); ***: p< 0.001 compared to negative control; *: p < 0.05 compared to negativecontrol.
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f07: : interleukin-8 (IL-8) production by envelope glycoprotein 2(E2)-stimulated human umbilical vein endothelial cells. Results presented asmean and standard deviation. The experiments were performed in triplicate. c-:cells and medium and phosphate-buffered saline (pH 7.2). LPS:lipopolysaccharide (1.0 µg/mL); Sup: culture supernatant Escherichiacoli BL21; TNF-α: tumour necrosis factor alpha (10 ng/mL); ***: p< 0.001 compared to negative control; *: p < 0.05 compared to negativecontrol.

Mentions: The E2 proteins were capable of inducing the production of IL-8 compared withnon-stimulated cells. The detection of IL-8 production by HUVECs is presented in Fig. 7. There was a statistically significantdifference (p < 0.05) between all of the stimuli tested compared with the negativecontrol. Unlike the IL-8 results, the E2 proteins were not able to induce the productionof TNF-α or LPS by HUVECs. However, 0.50 µM PMA induced HUVECs to produce 173.05 pg/mLTNF-α.


Inflammatory response of endothelial cells to hepatitis C virus recombinant envelope glycoprotein 2 protein exposure.

Urbaczek AC, Ribeiro LC, Ximenes VF, Afonso A, Nogueira CT, Generoso WC, Alberice JV, Rudnicki M, Ferrer R, Fonseca LM, Costa PI - Mem. Inst. Oswaldo Cruz (2014)

: interleukin-8 (IL-8) production by envelope glycoprotein 2(E2)-stimulated human umbilical vein endothelial cells. Results presented asmean and standard deviation. The experiments were performed in triplicate. c-:cells and medium and phosphate-buffered saline (pH 7.2). LPS:lipopolysaccharide (1.0 µg/mL); Sup: culture supernatant Escherichiacoli BL21; TNF-α: tumour necrosis factor alpha (10 ng/mL); ***: p< 0.001 compared to negative control; *: p < 0.05 compared to negativecontrol.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4238766&req=5

f07: : interleukin-8 (IL-8) production by envelope glycoprotein 2(E2)-stimulated human umbilical vein endothelial cells. Results presented asmean and standard deviation. The experiments were performed in triplicate. c-:cells and medium and phosphate-buffered saline (pH 7.2). LPS:lipopolysaccharide (1.0 µg/mL); Sup: culture supernatant Escherichiacoli BL21; TNF-α: tumour necrosis factor alpha (10 ng/mL); ***: p< 0.001 compared to negative control; *: p < 0.05 compared to negativecontrol.
Mentions: The E2 proteins were capable of inducing the production of IL-8 compared withnon-stimulated cells. The detection of IL-8 production by HUVECs is presented in Fig. 7. There was a statistically significantdifference (p < 0.05) between all of the stimuli tested compared with the negativecontrol. Unlike the IL-8 results, the E2 proteins were not able to induce the productionof TNF-α or LPS by HUVECs. However, 0.50 µM PMA induced HUVECs to produce 173.05 pg/mLTNF-α.

Bottom Line: 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.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Imunologia Clínica, Departamento de Análises Clínicas, Escola de Ciências Farmacêuticas, Bauru, SP, Brasil.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus