1. Hepatitis C virus production requires apolipoprotein A-I and affects its association with nascent low-density lipoproteins
- Author
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Leopoldo Paolo Pucillo, Corinna Steindler, Maria Antonella Longo, Carmine Mancone, Laura Amicone, Tonino Alonzi, Laura Santangelo, Marco Tripodi, Giacoma Simonte, Gianpiero D'Offizi, Cristina Di Giacomo, Chrysoula Vlassi, L. Spallanzani National Institute for Infectious Diseases, IRCCS, Department of Cellular Biotechnologies and Haematology, Institut Pasteur, Fondation Cenci Bolognetti - Istituto Pasteur Italia, Fondazione Cenci Bolognetti, and Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Università degli Studi di Roma 'La Sapienza' = Sapienza University [Rome]
- Subjects
Male ,Proteomics ,MESH: Lipoproteins, LDL ,Very low-density lipoprotein ,Apolipoprotein B ,hcv infection ,lipoproteins ,MESH: Apolipoprotein A-I ,Hepacivirus ,medicine.disease_cause ,Virus Replication ,MESH: Down-Regulation ,0302 clinical medicine ,MESH: Hepacivirus ,Electrophoresis, Gel, Two-Dimensional ,Cells, Cultured ,0303 health sciences ,MESH: Middle Aged ,biology ,medicine.diagnostic_test ,MESH: Proteomics ,Gastroenterology ,Middle Aged ,MESH: Case-Control Studies ,Hepatitis C ,3. Good health ,Lipoproteins, LDL ,030220 oncology & carcinogenesis ,[SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology ,MESH: Virion ,lipids (amino acids, peptides, and proteins) ,Female ,MESH: Cells, Cultured ,Adult ,Hepatitis C virus ,Down-Regulation ,03 medical and health sciences ,Viral life cycle ,Western blot ,Downregulation and upregulation ,medicine ,Humans ,030304 developmental biology ,MESH: Hepatitis C ,MESH: Humans ,Apolipoprotein A-I ,MESH: Virus Replication ,Virion ,MESH: Adult ,Lipid metabolism ,MESH: Electrophoresis, Gel, Two-Dimensional ,Virology ,Molecular biology ,MESH: Male ,Case-Control Studies ,biology.protein ,MESH: Female ,Lipoprotein - Abstract
Background/aims The life cycle of hepatitis C virus (HCV) is intimately linked to the lipid metabolism of the host. In particular, HCV exploits the metabolic machinery of the lipoproteins in several steps of its life cycle such as circulation in the bloodstream, cell attachment and entry, assembly and release of viral particles. However, the details of how HCV interacts with and influences the metabolism of the host lipoproteins are not well understood. A study was undertaken to investigate whether HCV directly affects the protein composition of host circulating lipoproteins. Methods A proteomic analysis of circulating very low-, low- and high-density lipoproteins (VLDL, LDL and HDL), isolated from either in-treatment naive HCV-infected patients or healthy donors (HD), was performed using two-dimensional gel electrophoresis and tandem mass spectrometry (MALDI-TOF/TOF). The results obtained were further investigated using in vitro models of HCV infection and replication. Results A decreased level of apolipoprotein A-I (apoA-I) was found in the LDL fractions of HCV-infected patients. This result was confirmed by western blot and ELISA analysis. HCV cellular models (JFH1 HCV cell culture system (HCVcc) and HCV subgenomic replicons) showed that the decreased apoA-I/LDL association originates from hepatic biogenesis rather than lipoprotein catabolism occurring in the circulation, and is not due to a downregulation of the apoA-I protein concentration. The sole non-structural viral proteins were sufficient to impair the apoA-I/LDL association. Functional evidence was obtained for involvement of apoA-I in the viral life cycle such as RNA replication and virion production. The specific siRNA-mediated downregulation of apoA-I led to a reduction in both HCV RNA and viral particle levels in culture. Conclusions This study shows that HCV induces lipoprotein structural modification and that its replication and production are linked to the host lipoprotein metabolism, suggesting apoA-I as a new possible target for antiviral therapy.
- Published
- 2010
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