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

Miriam Triyatni, Bertrand Saunier, Padma Maruvada, Anthony R. Davis, Luca Ulianich, Theo Heller, Arvind Patel, Leonard D. Kohn, T. Jake Liang

Research output: Contribution to journalArticlepeer-review

107 Scopus citations

Abstract

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/cm3 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 (Kd ∼1 μg/ml) and low (Kd ∼50 to 60 μg/ml) affinities of binding. Anti-E1 and -E2 antibodies inhibited HCV-LPs binding to cells. While preincubatioh 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.

Original languageEnglish (US)
Pages (from-to)9335-9344
Number of pages10
JournalJournal of Virology
Volume76
Issue number18
DOIs
StatePublished - Sep 2002
Externally publishedYes

ASJC Scopus subject areas

  • Immunology

Fingerprint

Dive into the research topics of 'Interaction of hepatitis C virus-like particles and cells: A model system for studying viral binding and entry'. Together they form a unique fingerprint.

Cite this