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Abstract

A better understanding of the antibody response during natural infection and the effect on disease progression and reinfection is necessary for the development of a protective hepatitis C virus (HCV) vaccine. The HCV pseudoparticle (HCVpp) system enables the study of viral entry and inhibition by antibody neutralization. A robust and comparable neutralization assay is crucial for the development and evaluation of experimental vaccines.

With the aim of optimizing the HCVpp–murine leukaemia virus (MLV) system, we tested the neutralization of HCVpp-harbouring E1E2 from 21 HCV isolates representing 6 different genotypes by several monoclonal antibodies (mAbs). HCVpps are generated by expressing functional envelope glycoproteins (E1E2) onto pseudoparticles derived from env-deleted MLV. Adjustments of E1E2, gag–pol and luciferase plasmid ratios resulted in increased yields for most HCVpps and recovery of one non-infectious HCVpp. We simplified and improved the protocol to achieve higher signal/noise ratios and minimized the amount of HCVpps and mAbs needed for the detection of neutralization. Using our optimized protocol, we demonstrated comparable results to previously reported data with both diluted and freeze–thawed HCVpps.

In conclusion, we successfully established a simplified and reproducible HCVpp neutralization protocol for studying a wide range of HCV variants. This simplified protocol provides highly consistent results and could be easily adopted by others to evaluate precious biological material. This will contribute to a better understanding of the antibody response during natural infection and help evaluate experimental HCV vaccines.

Funding
This study was supported by the:
  • Amsterdam University Medical Centers (Award PhD Scholarship 2016)
    • Principle Award Recipient: AnaLuz Chumbe Mendoza
  • Amsterdam University Medical Centers (Award Fellowship 2017)
    • Principle Award Recipient: MaritJ van Gils
  • Nederlandse Organisatie voor Wetenschappelijk Onderzoek (Award 015.015.042)
    • Principle Award Recipient: JankeSchinkel
  • Nederlandse Organisatie voor Wetenschappelijk Onderzoek (Award 91719372)
    • Principle Award Recipient: JankeSchinkel
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License.
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2022-11-18
2024-03-29
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References

  1. WHO guidelines Guidelines for the Care and Treatment of Persons Diagnosed with Chronic Hepatitis C Virus Infection 2018
    [Google Scholar]
  2. World Health Organization. n.d https://www.who.int/news-room/fact-sheets/detail/hepatitis-c accessed 18 October 2019
  3. WHO World Health Organization. WHO | Hepatitis C. n.d https://www.who.int/news-room/fact-sheets/detail/hepatitis-C accessed 11 November 2022
  4. Division of viral hepatitis, national center for HIV, viral hepatitis, STD and TP. Viral Hepatitis Surveillance Report 2019
    [Google Scholar]
  5. World Health Organization Global health sector strategy on viral hepatitis 2016-2021. Towards ending viral hepatitis. Glob Hepat Program Dep HIV/AIDS 2016
    [Google Scholar]
  6. Bailey JR, Barnes E, Cox AL. Approaches, progress, and challenges to hepatitis C vaccine development. Gastroenterology 2019; 156:418–430 [View Article]
    [Google Scholar]
  7. Ott DE. Cellular proteins in HIV virions. Rev Med Virol 1997; 7:167–180 [View Article]
    [Google Scholar]
  8. Millet JK, Whittaker GR. Murine leukemia virus (MLV)-based coronavirus spike-pseudotyped particle production and infection. Bio Protoc 2016; 6:1–18 [View Article]
    [Google Scholar]
  9. Ou W, Delisle J, Jacques J, Shih J, Price G et al. Induction of ebolavirus cross-species immunity using retrovirus-like particles bearing the Ebola virus glycoprotein lacking the mucin-like domain. Virol J 2012; 9:1–13 [View Article] [PubMed]
    [Google Scholar]
  10. Temperton NJ, Hoschler K, Major D, Nicolson C, Manvell R et al. A sensitive retroviral pseudotype assay for influenza H5N1-neutralizing antibodies. Influenza Other Respir Viruses 2007; 1:105–112 [View Article]
    [Google Scholar]
  11. Zheng Y, Larragoite ET, Williams ESCP, Lama J, Cisneros I et al. Neutralization assay with SARS-CoV-1 and SARS-CoV-2 spike pseudotyped murine leukemia virions. Virol J 2021; 18:1–6 [View Article] [PubMed]
    [Google Scholar]
  12. Nie J, Huang W, Liu Q, Wang Y. HIV-1 pseudoviruses constructed in China regulatory laboratory. Emerg Microbes Infect 2020; 9:32–41 [View Article] [PubMed]
    [Google Scholar]
  13. Bartosch B, Dubuisson J, Cosset F-L. Infectious hepatitis C virus pseudo-particles containing functional E1-E2 envelope protein complexes. J Exp Med 2003; 197:633–642 [View Article]
    [Google Scholar]
  14. Hsu M, Zhang J, Flint M, Logvinoff C, Cheng-Mayer C et al. Hepatitis C virus glycoproteins mediate pH-dependent cell entry of pseudotyped retroviral particles. Proc Natl Acad Sci U S A 2003; 100:7271–7276 [View Article] [PubMed]
    [Google Scholar]
  15. Wakita T, Pietschmann T, Kato T, Date T, Miyamoto M et al. Production of infectious hepatitis C virus in tissue culture from a cloned viral genome. Nat Med 2005; 11:791–796 [View Article] [PubMed]
    [Google Scholar]
  16. Zhong J, Gastaminza P, Cheng G, Kapadia S, Kato T et al. Robust hepatitis C virus infection in vitro. Proc Natl Acad Sci U S A 2005; 102:9294–9299 [View Article] [PubMed]
    [Google Scholar]
  17. Meng C, Qiu X, Jin S, Yu S, Chen H et al. Whole genome sequencing and biological characterization of Duck/JS/10, a new lentogenic class I Newcastle disease virus. Arch Virol 2012; 157:869–880 [View Article] [PubMed]
    [Google Scholar]
  18. Yang D, Zuo C, Wang X, Meng X, Xue B et al. Complete replication of hepatitis B virus and hepatitis C virus in a newly developed hepatoma cell line. Proc Natl Acad Sci U S A 2014; 111:E1264–73 [View Article]
    [Google Scholar]
  19. Wasilewski LN, Ray SC, Bailey JR. Hepatitis C virus resistance to broadly neutralizing antibodies measured using replication-competent virus and pseudoparticles. J Gen Virol 2016; 97:2883–2893 [View Article] [PubMed]
    [Google Scholar]
  20. Osburn WO, Snider AE, Wells BL, Latanich R, Bailey JR et al. Clearance of hepatitis C infection is associated with the early appearance of broad neutralizing antibody responses. Hepatology 2014; 59:2140–2151 [View Article]
    [Google Scholar]
  21. Urbanowicz RA, McClure CP, King B, Mason CP, Ball JK et al. Novel functional hepatitis C virus glycoprotein isolates identified using an optimized viral pseudotype entry assay. J Gen Virol 2016; 97:2265–2279 [View Article]
    [Google Scholar]
  22. Lavillette D, Tarr AW, Voisset C, Donot P, Bartosch B et al. Characterization of host-range and cell entry properties of the major genotypes and subtypes of hepatitis C virus. Hepatology 2005; 41:265–274 [View Article]
    [Google Scholar]
  23. Bailey JR, Urbanowicz RA, Ball JK, Law M, Foung SKH. Standardized Method for the Study of Antibody Neutralization of HCV Pseudoparticles (HCVpp). Methods Mol Biol 2019; 1911:441–450 [View Article]
    [Google Scholar]
  24. Kalemera MD, Capella-Pujol J, Chumbe A, Underwood A, Bull RA et al. Optimized cell systems for the investigation of hepatitis C virus E1E2 glycoproteins. J Gen Virol 2021; 102: [View Article]
    [Google Scholar]
  25. Urbanowicz RA, McClure CP, Brown RJP, Tsoleridis T, Persson MAA et al. A diverse panel of hepatitis C virus glycoproteins for use in vaccine research reveals extremes of monoclonal antibody neutralization resistance. J Virol 2015; 90:3288–3301 [View Article]
    [Google Scholar]
  26. Sarzotti-Kelsoe M, Bailer RT, Turk E, Lin C, Bilska M et al. Optimization and validation of the TZM-bl assay for standardized assessments of neutralizing antibodies against HIV-1. J Immunol Methods 2014; 409:131–146 [View Article] [PubMed]
    [Google Scholar]
  27. Oguntuyo KY, Stevens CS, Hung CT, Ikegame S, Acklin JA et al. Quantifying absolute neutralization titers against SARS-CoV-2 by a standardized virus neutralization assay allows for cross-cohort comparisons of COVID-19 sera. mBio 2021; 12:1–23 [View Article]
    [Google Scholar]
  28. Nie J, Li Q, Wu J, Zhao C, Hao H et al. Establishment and validation of a pseudovirus neutralization assay for SARS-CoV-2. Emerg Microbes Infect 2020; 9:680–686 [View Article] [PubMed]
    [Google Scholar]
  29. Johnson MC, Lyddon TD, Suarez R, Salcedo B, LePique M et al. Optimized pseudotyping conditions for the SARS-COV-2 spike glycoprotein. J Virol 2020; 94:1–10 [View Article]
    [Google Scholar]
  30. Merat SJ, Bru C, van de Berg D, Molenkamp R, Tarr AW et al. Cross-genotype AR3-specific neutralizing antibodies confer long-term protection in injecting drug users after HCV clearance. J Hepatol 2019; 71:14–24 [View Article] [PubMed]
    [Google Scholar]
  31. Meunier J-C, Russell RS, Goossens V, Priem S, Walter H et al. Isolation and characterization of broadly neutralizing human monoclonal antibodies to the E1 glycoprotein of hepatitis C virus. J Virol 2008; 82:966–973 [View Article] [PubMed]
    [Google Scholar]
  32. Giang E, Dorner M, Prentoe JC, Dreux M, Evans MJ et al. Human broadly neutralizing antibodies to the envelope glycoprotein complex of hepatitis C virus. Proc Natl Acad Sci U S A 2012; 109:6205–6210 [View Article] [PubMed]
    [Google Scholar]
  33. Clayton RF, Owsianka A, Aitken J, Graham S, Bhella D et al. Analysis of Antigenicity and Topology of E2 Glycoprotein Present on Recombinant Hepatitis C Virus-Like Particles. J Virol 2002; 76:9562 [View Article]
    [Google Scholar]
  34. Keck Z, Xia J, Wang Y, Wang W, Krey T et al. Human monoclonal antibodies to a novel cluster of conformational epitopes on HCV E2 with resistance to neutralization escape in a genotype 2a isolate. PLoS Pathog 2012; 8:e1002653 [View Article]
    [Google Scholar]
  35. Azzoni AR, Ribeiro SC, Monteiro GA, Prazeres DMF. The impact of polyadenylation signals on plasmid nuclease-resistance and transgene expression. J Gene Med 2007; 9:392–402 [View Article]
    [Google Scholar]
  36. Salas JH, Urbanowicz RA, Guest JD, Frumento N, Figueroa A et al. An antigenically diverse, representative panel of envelope glycoproteins for hepatitis C virus vaccine development. Gastroenterology 2022; 162:562–574 [View Article]
    [Google Scholar]
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