Hepatitis C virus resistance to broadly neutralizing antibodies measured using replication-competent virus and pseudoparticles

Lisa N. Wasilewski, Stuart C. Ray, Justin R. Bailey

Research output: Contribution to journalArticle

Abstract

A better understanding of natural variation in neutralization resistance and fitness of diverse hepatitis C virus (HCV) envelope (E1E2) variants will be critical to guide rational development of an HCV vaccine. This work has been hindered by inadequate genetic diversity in viral panels and by a lack of standardization of HCV entry assays. Neutralization assays generally use lentiviral pseudoparticles expressing HCV envelope proteins (HCVpp) or chimeric full-length viruses that are replication competent in cell culture (HCVcc). There have been few systematic comparisons of specific infectivities of E1E2-matched HCVcc and HCVpp, and to our knowledge, neutralization of E1E2-matched HCVpp and HCVcc has never been compared using a diverse panel of human broadly neutralizing monoclonal antibodies (bNAbs) targeting distinct epitopes. Here, we describe an efficient method for introduction of naturally occurring E1E2 genes into a full-length HCV genome, producing replication-competent chimeric HCVcc. We generated diverse panels of E1E2-matched HCVcc and HCVpp and measured the entry-mediating fitness of E1E2 variants using the two systems. We also compared neutralization of E1E2-matched HCVcc and HCVpp by a diverse panel of human bNAbs targeting epitopes across E1E2. We found no correlation between specific infectivities of E1E2-matched HCVcc versus HCVpp, but found a very strong positive correlation between relative neutralization resistance of these same E1E2-matched HCVcc and HCVpp variants. These results suggest that quantitative comparisons of neutralization resistance of E1E2 variants can be made with confidence using either HCVcc or HCVpp, allowing the use of either or both systems to maximize diversity of neutralization panels.

Original languageEnglish (US)
Article number000608
Pages (from-to)2883-2893
Number of pages11
JournalJournal of General Virology
Volume97
Issue number11
DOIs
StatePublished - Nov 1 2016

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Virus Replication
Neutralizing Antibodies
Hepacivirus
Proteins
Epitopes
Monoclonal Antibodies
Viral Envelope Proteins
Virus Internalization
Vaccines
Cell Culture Techniques
Genome
Genes

Keywords

  • Broadly neutralizing antibody
  • HCVcc
  • Hepatitis C virus
  • Pseudoparticle

ASJC Scopus subject areas

  • Virology

Cite this

Hepatitis C virus resistance to broadly neutralizing antibodies measured using replication-competent virus and pseudoparticles. / Wasilewski, Lisa N.; Ray, Stuart C.; Bailey, Justin R.

In: Journal of General Virology, Vol. 97, No. 11, 000608, 01.11.2016, p. 2883-2893.

Research output: Contribution to journalArticle

Wasilewski, Lisa N.; Ray, Stuart C.; Bailey, Justin R. / Hepatitis C virus resistance to broadly neutralizing antibodies measured using replication-competent virus and pseudoparticles.

In: Journal of General Virology, Vol. 97, No. 11, 000608, 01.11.2016, p. 2883-2893.

Research output: Contribution to journalArticle

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