Method of detecting antibodies to hcv

Abstract

A family of cDNA sequences derived from hepatitis C virus (HCV) are provided. These sequences encode antigens which react immunologically with antibodies present in individuals with non-A non-B hepatitis (NANBH), but which are absent from individuals infected with hepatitis A virus, or hepatitis B virus, and also are absent in control individuals. The HCV cDNA sequences lack substantial homology to the sequences of hepatitis delta virus (HDV) and HBV. A comparison of the sequences of amino acids encoded in the HCV cDNA with the sequences of Flaviviruses indicates that HCV may be related to the Flaviviruses. The HCV cDNA sequences and the polypeptides encoded therein are useful as reagents for the detection and therapy of HCV. The reagents provided in the invention are also useful for the isolation of NANBH agent(s), for the propagation of these agents in tissue culture, and for the screening of antiviral agents for HCV.

1 . A purified HCV polynucleotide.  
     
     
         2 . A recombinant HCV polynucleotide.  
     
     
         3 . A recombinant polynucleotide comprising a sequence derived from an HCV genome or from HCV cDNA.  
     
     
         4 . A recombinant polynucleotide encoding an epitope of HCV.  
     
     
         5 . A recombinant vector containing the polynucleotide of  claim 2 , or  claim 3 , or  claim 4 .  
     
     
         6 . A host cell transformed with the vector of  claim 5 .  
     
     
         7 . A recombinant expression system comprising an open reading frame (ORF) of DNA derived from an HCV genome or from HCV cDNA, wherein the ORF is operably linked to a control sequence compatible with a desired host.  
     
     
         8 . A cell transformed with the recombinant expression system of  claim 7 .  
     
     
         9 . A polypeptide produced by the cell of  claim 8 .  
     
     
         10 . Purified HCV.  
     
     
         11 . A preparation of polypeptides from the HCV of  claim 10 .  
     
     
         12 . A purified HCV polypeptide.  
     
     
         13 . A purified polypeptide comprising an epitope which is immunologically identifiable with an epitope contained in HCV.  
     
     
         14 . A recombinant HCV polypeptide.  
     
     
         15 . A recombinant polypeptide comprised of a sequence derived from an HCV genome or from HCV cDNA.  
     
     
         16 . A recombinant polypeptide comprised of an HCV epitope.  
     
     
         17 . A fusion polypeptide comprised of an HCV polypeptide.  
     
     
         18 . A monoclonal antibody directed against an HCV epitope.  
     
     
         19 . A purified preparation of polyclonal antibodies directed against HCV.  
     
     
         20 . A particle which is immunogenic against HCV infection comprising a non-HCV polypeptide having an amino acid sequence capable of forming a particle when said sequence is produced in a eukaryotic host, and an HCV epitope.  
     
     
         21 . A polynucleotide probe for HCV.  
     
     
         22 . A kit for analyzing samples for the presence of polynucleotides derived from HCV comprising a polynucleotide probe containing a nucleotide sequence from HCV of about 8 or more nucleotides, in a suitable container.  
     
     
         23 . A kit for analyzing samples for the presence of an HCV antigen comprising an antibody directed against the HCV antigen to be detected, in a suitable container.  
     
     
         24 . A kit for analyzing samples for the presence of an antibodies directed against an HCV antigen comprising a polypeptide containing an HCV epitope present in the HCV antigen, in a suitable container.  
     
     
         25 . A polypeptide comprised of an HCV epitope, attached to a solid substrate.  
     
     
         26 . An antibody to an HCV epitope, attached to a solid substrate.  
     
     
         27 . A method for producing a polypeptide containing an HCV epitope comprising incubating host cells transformed with an expression vector containing a sequence encoding a polypeptide containing an HCV epitope under conditions which allow expression of said polypeptide.  
     
     
         28 . A polypeptide containing an HCV epitope produced by the method of  claim 27 .  
     
     
         29 . A method for detecting HCV nucleic acids in a sample comprising: 
 (a) reacting nucleic acids of the sample with a probe for an HCV polynucleotide under conditions which allow the formation of a polynucleotide duplex between the probe and the HCV nucleic acid from the sample; and    (b) detecting a polynucleotide duplex which contains the probe.    
     
     
         30 . An immunoassay for detecting an HCV antigen comprising: 
 (a) incubating a sample suspected of containing an HCV antigen with a probe antibody directed against the HCV antigen to be detected under conditions which allow the formation of an antigen-antibody complex; and    (b) detecting an antigen-antibody complex containing the probe antibody.    
     
     
         31 . An immunoassay for detecting antibodies directed against an HCV antigen comprising: 
 (a) incubating a sample suspected of containing anti-HCV antibodies with a probe polypeptide which contains an epitope of the HCV, under conditions which allow the formation of an antibody-antigen complex; and    (b) detecting the antibody-antigen complex containing the probe antigen.    
     
     
         32 . A vaccine for treatment of HCV infection comprising an immunogenic polypeptide containing an HCV epitope wherein the immunogenic polypeptide is present in a pharmacologically effective dose in a pharmaceutically acceptable excipient.  
     
     
         33 . A vaccine for treatment of HCV infection comprising inactivated HCV in a pharmacologically effective dose in a pharmaceutically acceptable excipient.  
     
     
         34 . A vaccine for treatment of HCV infection comprising attenuated HCV in a pharmacologically effective dose in a pharmaceutically acceptable excipient.  
     
     
         35 . A tissue culture grown cell infected with HCV.  
     
     
         36 . The HCV infected cell of  claim 35 , wherein the cell is of a human macrophage cell line, or is of a hepatocyte cell line, or is of a mosquito cell line, or is of a tick cell line, or is of a mouse macrophage cell line, or is an embryonic cell.  
     
     
         37 . The HCV infected cell of  claim 35 , wherein the cell is of a cell line derived from liver of an HCV infected individual.  
     
     
         38 . A method for producing antibodies to HCV comprising administering to an individual an isolated immunogenic polypeptide containing an HCV epitope in an amount sufficient to produce-an immune response.  
     
     
         39 . A method for producing antibodies to HCV comprising administering to an individual the polypeptide preparation of  claim 11 , wherein the preparation contains at least 1 immunogenic polypeptide, and the administering is of an amount sufficient to produce an immune response.