Localization and reactivity of an immunodominant domain in the NS3 region of hepatitis C virus.
Analysis of the amino acid sequences of the nonstructural region 3 (NS3) of the hepatitis C virus type 1 revealed four points with a high average hydrophilicity (Ah). Two of these potential antigenic sites were expressed in E. coli as short fragments. The first fragment of 91 residues (NS3f3: residues 1359-1449) harbors the hexapeptide K-K-K-C-D-E with an Ah of 2.33; the second fragment is 73 residues long (NS3f4: residues 1460-1532) and encompasses the heptapeptide R-S-N-R-R-G-R with an Ah of 1.79. Both fragments were expressed with truncated hepatitis B core (tHBc) as a carrier protein. The fusion proteins were purified from the bacterial lysates by affinity chromatography on immobilized monoclonal antibodies against HBc, and evaluated as antigens in an enzyme immunoassay for the detection of HCV antibodies. In a specificity control panel, reactivity with NS3f3 was only found in proven HCV carriers, while reactivity with NS3f4 was weak in HCV carriers but accounted for some of the nonspecific serological reactions. In a group of 48 genotyped HCV-infected volunteer blood donors, antibodies against NS3f3 were detected in 90% (27/30) of HCV-type 1 infections and in all HCV-type 4 infections (5/5).(ABSTRACT TRUNCATED AT 250 WORDS)
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Belgian Red Cross Blood Transfusion Center, Leuven, Belgium.
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D000595:Amino Acid Sequence; D000956:Antigens, Viral / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004279:DNA, Viral / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D016174:Hepacivirus / Q000235:genetics* / Q000276:immunology*; D006508:Hepatitis Antibodies / Q000097:blood; D006526:Hepatitis C / Q000175:diagnosis / Q000276:immunology; D018937:Hepatitis C Antibodies; D018936:Hepatitis C Antigens; D006801:Humans; D016056:Immunodominant Epitopes / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D017361:Viral Nonstructural Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*
D000956:Antigens, Viral; D017931:DNA Primers; D004279:DNA, Viral; D006508:Hepatitis Antibodies; D018937:Hepatitis C Antibodies; D018936:Hepatitis C Antigens; D016056:Immunodominant Epitopes; D011993:Recombinant Fusion Proteins; D017361:Viral Nonstructural Proteins