Interaction of hepatitis C virus-like particles and cells: a model system for studying viral binding and entry.

  • Triyatni M
  • Saunier B
  • Maruvada P
 et al. 
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Hepatitis C virus-like particles (HCV-LPs) containing the structural proteins of HCV H77 strain (1a genotype) was used as a model for HCV virion to study virus-cell interaction. HCV-LPs showed a buoyant density of 1.17 to 1.22 g/cm(3) in a sucrose gradient and formed double-shelled particles 35 to 49 nm in diameter. Flow cytometry analysis by an indirect method (detection with anti-E2 antibody) and a direct method (use of dye-labeled HCV-LPs) showed that HCV-LPs binds to several human hepatic (primary hepatocytes, HepG2, HuH7, and NKNT-3) and T-cell (Molt-4) lines. HCV-LPs binding to cells occurred in a dose- and calcium-dependent manner and was not mediated by CD81. Scatchard plot analysis suggests the presence of two binding sites for HCV-LPs with high (K(d) approximately 1 microg/ml) and low (K(d) approximately 50 to 60 microg/ml) affinities of binding. Anti-E1 and -E2 antibodies inhibited HCV-LPs binding to cells. While preincubation of HCV-LPs with very-low-density lipoprotein (VLDL), low-density lipoprotein (LDL), or high-density lipoprotein (HDL) blocked its binding to cells, preincubation of cells with VLDL, LDL, HDL, or anti-LDL-R antibody did not. Confocal microscopy analysis showed that, after binding to cells, dye-labeled HCV-LPs were internalized into the cytoplasm. This process could be inhibited with anti-E1 or anti-E2 antibodies, suggesting that E1 and E2 proteins mediate HCV-LPs binding and, subsequently, their entry into cells. Altogether, our results indicate that HCV-LPs can be used to further characterize the mechanisms involved in the early steps of HCV infection.

Author-supplied keywords

  • Antigens, CD
  • Antigens, CD81
  • Antigens, CD: metabolism
  • Carcinoma, Hepatocellular
  • Cell Line
  • Hepacivirus
  • Hepacivirus: physiology
  • Hepatocytes
  • Hepatocytes: virology
  • Humans
  • Lipoproteins
  • Lipoproteins: pharmacology
  • Membrane Fusion
  • Membrane Proteins
  • Microscopy, Confocal
  • T-Lymphocytes
  • T-Lymphocytes: virology
  • Tumor Cells, Cultured
  • Viral Envelope Proteins
  • Viral Envelope Proteins: metabolism
  • Virion
  • Virion: physiology

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  • Miriam Triyatni

  • Bertrand Saunier

  • Padma Maruvada

  • Anthony R Davis

  • Luca Ulianich

  • Theo Heller

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