HCV REPLICATION AND IN VITRO NEUTRALIZATION ASSAY

  • Lanford, Robert E (PI)

    Project: Research project

    Project Details

    Description

    Hepatitis C virus (HCV) infections represent a worldwide health problem,
    with approximately 0.5 to 1.5% of most populations testing positive for
    antibodies to HCV. An estimated 50% of infected individuals will become
    chronic carriers of HCV and will be at increased risk for the development
    of cirrhosis and hepatocellular carcinoma and will also act as a source
    of infection for others. Transmission of HCV occurs most readily through
    contact with contaminated blood as in blood transfusions and IV drug
    abuse; however, the true epidemiology of this agent is only now emerging
    through the use of diagnostic assays. Other routes of transmission
    include sexual contact, intrafamilial contact, and transmission from
    mother to child at birth. The development of a vaccine for this virus
    must be considered a high priority. Currently, the only available method
    of testing vaccine candidates is the immunization and challenge of
    chimpanzees, a very costly and time consuming approach. The development
    of an in vitro assay for evaluating the presence of neutralizing
    antibodies would greatly facilitate vaccine development. This proposal
    will employ a recently developed primary hepatocyte tissue culture system
    for the in vitro propagation of HCV to address these needs. The specific
    aims of this proposal are: 1) to optimize methods for detecting HCV
    polypeptides using a human hepatoma cell line transfected with
    recombinant expression vectors encoding various domains of the HCV
    polyprotein; 2) to produce antibodies to the capsid and envelope proteins
    of HCV expressed in the insect cell/baculovirus system; 3) to define the
    authentic HCV proteins expressed in HCV-infected primary hepatocytes
    using immunoprecipitation and immunoblot methodology; 4) to develop a
    quantitative assay for measuring in vitro infections of primary
    hepatocytes with HCV and to use this assay to develop an in vitro
    neutralization assay for HCV; 5) to examine panels of human and
    chimpanzee sera for the presence of neutralizing antibodies and to
    evaluate antisera produced against the envelope proteins of HCV for
    neutralizing activity; and 6) to create immortalized human and chimpanzee
    hepatocyte cell lines permissive for HCV infections.
    StatusFinished
    Effective start/end date9/1/928/31/95

    Funding

    • National Institutes of Health: $221,790.00
    • National Institutes of Health

    ASJC

    • Medicine(all)

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