Gastroenterology

Gastroenterology

Volume 132, Issue 2, February 2007, Pages 667-678
Gastroenterology

Basic–liver, pancreas, and biliary tract
Hepatitis C Virus Continuously Escapes From Neutralizing Antibody and T-Cell Responses During Chronic Infection In Vivo

https://doi.org/10.1053/j.gastro.2006.12.008Get rights and content

Background & Aims: Broadly reactive neutralizing antibodies (nAbs) and multispecific T-cell responses are generated during chronic hepatitis C virus (HCV) infection and yet fail to clear the virus. This study investigated the development of autologous nAb and HCV-glycoprotein–specific T-cell responses and their effects on viral sequence evolution during chronic infection in order to understand the reasons for their lack of effectiveness.

Methods: Numerous E1E2 sequences were amplified and sequenced from serum samples collected over a 26-year period from patient H, a uniquely well-characterized, chronically infected individual. HCV pseudoparticles (HCVpp) expressing the patient-derived glycoproteins were generated and tested for their sensitivity to neutralization by autologous and heterologous serum antibodies.

Results: A strain-specific nAb response developed early in infection (8 weeks postinfection), whereas cross-reactive antibodies able to neutralize HCVpp-bearing heterologous glycoproteins developed late in infection (>33 wk postinfection). The humoral response continuously failed to neutralize viruses bearing autologous glycoprotein sequences that were present in the serum at a given time. The amplified glycoprotein sequences displayed high variability, particularly in regions corresponding to defined linear B-cell epitopes. Mutations in defined neutralizing epitopes were associated with a loss of recognition by monoclonal antibodies against these epitopes and with decreased neutralization of corresponding HCVpp. Viral escape from CD4 and CD8 T-cell responses also was shown for several novel epitopes throughout the glycoprotein region.

Conclusions: During chronic infection HCV is subjected to selection pressures from both humoral and cellular immunity, resulting in the continuous generation of escape variants.

Section snippets

Antibodies, Cells, and Reagents

Both 293T and Hep3B cells were propagated in Dulbecco’s modified Eagle medium with 10% fetal bovine serum. Antibodies against HCV E2 have been described previously.14, 23 To generate soluble CD81 an expression plasmid encoding the CD81 large extracellular loop fused to Glutathione S-transferase (GST) was transformed into Rosetta-gami Escherichia coli (Novagen, La Jolla, CA). Fusion proteins were prepared by lysis with an Avestin (Avestin, Ottawa, Canada) air emulsifier (3 passages at 15,000

Clinical Information

Patient H, the subject of this study, was infected with HCV through a blood transfusion in 1977 while undergoing cardiac surgery. After an initial high alanine aminotransferase peak (2112 IU/L), indicative of severe acute hepatitis, he went on to develop mild chronic hepatitis with persistently low alanine aminotransferase levels and HCV RNA levels in the 104 and 106 genome copies/mL range over the following 26 years (Figure 1A). Patient H underwent liver biopsy procedures on 4 occasions, each

Discussion

This study is an in-depth analysis of the nAb response against autologous HCV E1E2 sequences and shows ongoing evasion of the virus-specific adaptive immune response during persistent infection. Patient H is a uniquely well-suited subject for this study because he is representative of many patients with mild, chronic hepatitis C and clinically is well characterized with serum samples spanning a long period of time. Furthermore, specific research tools such as sequences, infectious clones,

References (59)

  • M. Alter et al.

    The natural history of community-acquired hepatitis C in the United States

    N Engl J Med

    (1992)
  • M. Martell et al.

    Hepatitis C virus (HCV) circulates as a population of different but closely related genomes: quasispecies nature of HCV genome distribution

    J Virol

    (1992)
  • R. Brown et al.

    Evolutionary dynamics of hepatitis C virus envelope genes during chronic infection

    J Gen Virol

    (2005)
  • A. Weiner et al.

    Persistent hepatitis C virus infection in a chimpanzee is associated with emergence of a cytotoxic T lymphocyte escape variant

    Proc Natl Acad Sci U S A

    (1995)
  • K. Chang et al.

    Immunological significance of cytotoxic T lymphocyte epitope variants in patients chronically infected by the hepatitis C virus

    J Clin Invest

    (1997)
  • U. Seifert et al.

    Hepatitis C virus mutation affects proteasomal epitope processing

    J Clin Invest

    (2004)
  • F. Lechner et al.

    Analysis of successful immune responses in persons infected with hepatitis C virus

    J Exp Med

    (2000)
  • R. Thimme et al.

    Determinants of viral clearance and persistence during acute hepatitis C virus infection

    J Exp Med

    (2001)
  • N.H. Shoukry et al.

    Memory CD8+ T cells are required for protection from persistent hepatitis C virus infection

    J Exp Med

    (2003)
  • A. Grakoui et al.

    HCV persistence and immune evasion in the absence of memory T cell help

    Science

    (2003)
  • M. Hsu et al.

    Hepatitis C virus glycoproteins mediate pH-dependent cell entry of pseudotyped retroviral particles

    Proc Natl Acad Sci U S A

    (2003)
  • B. Bartosch et al.

    In vitro assay for neutralizing antibody to hepatitis C virus: evidence for broadly conserved neutralization epitopes

    Proc Natl Acad Sci U S A

    (2003)
  • C. Logvinoff et al.

    Neutralizing antibody response during acute and chronic hepatitis C virus infection

    Proc Natl Acad Sci U S A

    (2004)
  • J. McKeating et al.

    Diverse hepatitis C virus glycoproteins mediate viral infection in a CD81-dependent manner

    J Virol

    (2004)
  • J.C. Meunier et al.

    Evidence for cross-genotype neutralization of hepatitis C virus pseudo-particles and enhancement of infectivity by apolipoprotein C1

    Proc Natl Acad Sci U S A

    (2005)
  • M. Yanagi et al.

    Transcripts from a single full-length cDNA clone of hepatitis C virus are infectious when directly transfected into the liver of a chimpanzee

    Proc Natl Acad Sci U S A

    (1997)
  • A. Kolykhalov et al.

    Transmission of hepatitis C by intrahepatic inoculation with transcribed RNA

    Science

    (1997)
  • N. Ogata et al.

    Nucleotide sequence and mutation rate of the H strain of hepatitis C virus

    Proc Natl Acad Sci U S A

    (1991)
  • M. Flint et al.

    Characterization of hepatitis C virus E2 glycoprotein interaction with a putative cellular receptor, CD81

    J Virol

    (1999)
  • Cited by (0)

    Supported by PHS grants CA57973, AI50798, U19 AI40034, Medical Research Council, UK (G0400802), the National Institute of Diabetes and Digestive and Kidney Diseases intramural research program, the Greenberg Medical Research Institute, and the Starr Foundation. T.v.H. is supported by a postdoctoral fellowship from the Deutsche Forschungsgemeinschaft.

    View full text