Characterisation of the differences between hepatitis C virus genotype 3 and 1 glycoproteins

Authors

  • Megan L. Shaw,

    1. Division of Virology, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow, United Kingdom
    Current affiliation:
    1. Department of Microbiology, Mount Sinai School of Medicine, New York, NY 10029.
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  • John McLauchlan,

    1. Medical Research Council Virology Unit, Institute of Virology, Glasgow, United Kingdom
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  • Peter R. Mills,

    1. Gartnavel General Hospital, North Glasgow University Hospitals, NHS Trust, Glasgow, United Kingdom
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  • Arvind H. Patel,

    1. Medical Research Council Virology Unit, Institute of Virology, Glasgow, United Kingdom
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  • Elizabeth A.B. McCruden

    Corresponding author
    1. Division of Virology, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow, United Kingdom
    2. Gartnavel General Hospital, North Glasgow University Hospitals, NHS Trust, Glasgow, United Kingdom
    • Institute of Virology, Church Street, Glasgow G11 5JR, Scotland.
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Abstract

Sequence variation in the envelope E1 and E2 glycoproteins of hepatitis C virus (HCV) could account for differences in disease pathogenesis in patients infected with different genotypes. A cDNA encoding the structural region of the hepatitis C polyprotein was constructed to match the majority sequence of viral RNA extracted from a patient infected with genotype 3a (designated strain HCV3a-Gla). The principal differences predicted between E2 of HCV3a-Gla and the corresponding H77c genotype 1a protein were that the former contained six more amino acids (361 vs. 355), but it had one fewer glycosylation site. Expression studies showed that, in common with the H77c glycoproteins, E1 and E2 from HCV3a-Gla localised to the endoplasmic reticulum (ER) membrane in both Huh-7 and BHK tissue culture cells and interacted to form native complexes. Analysis of the cross-reactivity of antibodies raised against glycoproteins of genotype 1a strains showed that three of five monoclonal antibodies that recognise linear epitopes were able to detect E2 from strain HCV3a-Gla. However, neither conformational E2 antibodies nor antibodies raised against E1 were able to detect the HCV3a-Gla glycoproteins. In receptor binding assays, E2 of HCV3a-Gla consistently failed to bind CD81, a putative cell receptor for HCV. Absence of binding to CD81 and lack of recognition by most antibodies raised to genotype 1a glycoproteins indicate important differences between these glycoproteins representative of genotypes 3a and 1a. These may be pertinent to the differences in response to interferon therapy and the prevalence of steatosis reported in patients infected with these genotypes. J. Med. Virol. 70:361–372, 2003. © 2003 Wiley-Liss, Inc.

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