Subject(s)
Humans , Animals , Hepacivirus/genetics , Hepacivirus/metabolism , Viral Envelope Proteins/chemistry , Viral Envelope Proteins/genetics , Cell Line , Glycosylation , HeLa Cells , Hepatitis C Antibodies , Hepacivirus/immunology , Molecular Weight , Protein Processing, Post-Translational , Viral Envelope Proteins/metabolismABSTRACT
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.
Subject(s)
Humans , Hepacivirus/metabolism , Human Umbilical Vein Endothelial Cells/immunology , Human Umbilical Vein Endothelial Cells/pathology , Nitric Oxide/metabolism , Tumor Necrosis Factor-alpha/metabolism , Viral Envelope Proteins/metabolism , Apoptosis/genetics , Arginase/metabolism , Cell Survival , Escherichia coli/metabolism , Fibrosis , Gene Expression/genetics , Genetic Engineering/methods , Genetic Vectors/metabolism , Hepacivirus/immunology , Hepatitis C Antigens/metabolism , Inflammation/metabolism , /metabolism , Pichia/metabolism , Plasmids/metabolism , Recombinant Proteins , Vascular Endothelial Growth Factor A/metabolismABSTRACT
A infecção pelo vírus da hepatite C (VHC) é uma das mais comuns infecções ao redor do mundo. Aproximadamente, 20% dos pacientes infectados eliminam espontaneamente o vírus, porém a maioria dos indivíduos infectados desenvolve infecção crônica com amplo espectro de lesões hepáticas, desde inflamação leve até cirrose. A resposta imune do hospedeiro exerce grande influência sobre o desfecho da infecção pelo VHC. O objetivo deste trabalho foi analisar a influência dos polimorfismos genéticos de citocinas na susceptibilidade ou persistência da infecção por VHC e no clareamento espontâneo em uma amostra de pacientes da população do Rio de Janeiro (Brasil). Os polimorfismos genéticos das citocinas TNFA (-308), TGFB1 (codon 10 e 25), IL10 (-1082, -592), IL6 (-174) e IFNG (+874) foram analisados por PCR-SSP em 245 pacientes com hepatite C crônica (HCC), 41 pacientes que alcançaram o clareamento viral espontâneo e 189 indivíduos controle saudáveis. Além disso, os polimorfismos próximos ao gene da citocina IL28B (rs12979860, rs12980275 e rs8099917) foram analisados por PCR em tempo real em todos os grupos...
Hepatitis C virus infection is one of the most common blood-borne infections worldwide. Approximately, 20% of infected patients successfully eliminate the virus, whereas the majority of patients develop chronic infection with a wide spectrum of liver lesions, ranging from a minimal inflammation to cirrhosis. The host's immune response is an important correlate of HCV infection outcome and disease progression. The aim of this study was to explore the possibility of the inheritance of cytokine gene polymorphisms as a candidate for susceptibility to persistent HCV infection or HCV clearance in a sample of Rio de Janeiro (Brazil) population. Genetic polymorphisms in the cytokines TNFA (-308), TGFB1 (codon 10 and 25), IL10 (-1082, -592), IL6 (-174) and IFNG (+874) were analyzed by polymerase chain reaction-sequence-specific primer (SSP) in 245 chronic hepatitis C (HCC) patients, 41 spontaneous recovery (SR) patients and 189 healthy volunteers. Further, IL28B (rs12979860, rs12980275 and rs8099917) were assessed by real-time PCR in all groups...
Subject(s)
Humans , Male , Female , Liver Cirrhosis/therapy , Cytokines/analysis , Hepacivirus/metabolism , Hepatitis C, Chronic/pathology , Polymorphism, Genetic , Cytokines/genetics , Hepacivirus/pathogenicity , Hepatitis C, Chronic/complications , Heredity/geneticsABSTRACT
BACKGROUND & OBJECTIVE: India has a high prevalence of HIV-1, hapatitis C and B virus (HCV and HBV) in the blood donors but has yet to implement nucleic acid testing (NAT) in blood screening. We undertook a multicentre evaluation of blood donor testing by NAT for simultaneous detection of HIV-1, HBV and HCV in a single tube and also to determine the feasibility of NAT implementation in India's low volume setting. METHODS: A total of 12,224 unlinked samples along with their serological results were obtained from representative eight blood banks in India and were individually manually tested by the Procleix Ultrio Assay (Chiron Corp. Emeryville, CA) for simultaneous detection of HIV-1, HCV, and HBV. RESULTS: Of the 12,224 samples tested, 209 (1.71%) were seroreactive. One hundred thirty three samples (1.09%) were reactive by Ultrio assay, 84 samples were seroreactive but NAT non reactive. There were eight NAT yield cases: 1 HIV, 1 HIV-HCV co-infection, and 6 HBV. INTERPRETATION & CONCLUSION: Our observed NAT yield for all three viruses was 1 in 1528 (0.065%). We estimate NAT could interdict 3272 infectious donations a year among our approximate 5 million annual donations.
Subject(s)
Blood Banks , Blood Donors , Female , HIV Infections/diagnosis , HIV-1/metabolism , Hepacivirus/metabolism , Hepatitis B/diagnosis , Hepatitis B virus/metabolism , Hepatitis C/diagnosis , Humans , India , Male , Mass Screening/methods , Nucleic Acid Amplification Techniques/standards , RNA, Viral/analysis , Serologic Tests/standardsABSTRACT
O recente conhecimento das novas viroses hepatotrópicas, estimula o entendimento da história natural da hepatite induzida por estes agentes. Esse estudo objetivou a investigação clínica, bioquímica e sorológica, assim como a etiologia dos casos de hepatite aguda viral Não-A Não-B acompanhados num Centro de Referência para Doenças Hepáticas em Salvador-Bahia. Entre Julho/92 e Outubro/94, 147 pacientes com quadro clínico-laboratorial de hepatite aguda viral foram avaliados no Serviço de Gastro-Hepatologia. A suspeita de hepatite aguda viral se deu pelos achados clínicos clássicos da doença, assim como pela elevação da ALT superior a quatro vezes o limite máximo da normalidade. O VHA e VHB foram investigados através da determinação de anti-VHA IgM, anti-HBc IgM, AgHBs. Nos casos negativos para esses marcadores seguiuse a determinação de anti-CMV IgM, anti-EBV IgM, anti-VHC, antiVHE. O VHC-RNA por PCR foi realizado em todos os pacientes com hepatite Não-A Não-B e o VHG-RNA por PCR foi realizado nos pacientes negativos para o VHC-RNA. Dos 147 pacientes estudados, 53 (36,6%) foram relacionados ao VHA, 51 (34,7%) ao VHB, 13 (8,8%) ao VHC, 5 (3,4%) ao VHE, e 25 (17%) foram soro-negativos "pra todos os marcadores supracitados., (VHX). Nenhum desses pacientes foi positivo para EBV e CMV. No total, 43 pacientes foram considerados como hepatite aguda Não-A Não-B, sendo 5/43 (11,6%) pelo VHE, 13/43 (30,1%) pelo VHC, e 25/43 (58,1%) pelo VHX. Na "coorte" montada a partir do diagnóstico de hepatite Não-A Não-B, observamos que os 5 casos com soro-positividade para o VHE resolveram sua doença nos primeiros seis meses de acompanhamento. Nos pacientes com etiologia pelo VHC, 53,8°1 (7/13) normalizaram AL T nos primeiros seis meses de acompFnhamento, persistindo alterações ALT em 46,2% (6/13). O VHC-RNA permaneceu positivo após 6 meses de acompanhamento em 69,2% (9/13) pacientes sugerindo cronificação do processo independente dos valores de ALT...