Recombinant hepatitis C virus-like particles expressed by baculovirus: utility in cell-binding and antibody detection assays.
Hepatitis C virus (HCV) is difficult to study due to the lack of an efficient cell culture system or small animal model. As a result, HCV-cell interactions are not well-defined. In addition, several studies have identified a subset of patients in whom HCV RNA is present, but HCV antibody is not detected. We produced recombinant baculoviruses that expressed HCV structural proteins (core, E1 and E2, nt 342-2651) or control proteins. The HCV structural protein precursor was processed into immunoreactive proteins of appropriate size, and sucrose density sedimentation and electron microscopy of infected cell lysates demonstrated particle formation. To evaluate HCV antigenicity, particularly in patients who tested negative for HCV antibody in commercial HCV immunoassays but had persistent viremia, we evaluated the virus-like particles (VLPs) in solid-phase immunoassays. VLPs reacted with sera from HCV antibody positive subjects in these solid phase immunoassays, but not with control sera. Plasma samples from 19% (5/26) of HCV antibody negative subjects who were persistently HCV RNA positive also reacted with the HCV VLPs. When incubated with MOLT-4 cells at 4 degrees C, HCV VLPs demonstrated cell binding, and behaved similar to plasma-derived HCV preparations in a flow cytometry-based cell binding assay. These data suggest that recombinant HCV VLPs may allow identification of HCV antibody in patients, including some patients with persistent viremia and who are seronegative with current assays. In addition, HCV VLPs seem useful for evaluating HCV-cell interactions.
Xiang
Jinhua
J
Department of Internal Medicine, Iowa City Veterans Administration Medical Center, Iowa City, Iowa, USA.
Wünschmann
Sabina
S
George
Sarah L
SL
Klinzman
Donna
D
Schmidt
Warren N
WN
LaBrecque
Douglas R
DR
Stapleton
Jack T
JT
D000818:Animals; D000956:Antigens, Viral / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D016367:Baculoviridae / Q000235:genetics*; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D016174:Hepacivirus / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006526:Hepatitis C / Q000097:blood / Q000276:immunology; D018937:Hepatitis C Antibodies / Q000097:blood*; D006801:Humans; D007118:Immunoassay; D008854:Microscopy, Electron; D011994:Recombinant Proteins / Q000096:biosynthesis; D018411:Spodoptera / Q000821:virology; D014758:Viral Core Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics; D015678:Viral Structural Proteins / Q000096:biosynthesis / Q000235:genetics
D000956:Antigens, Viral; D018937:Hepatitis C Antibodies; D011994:Recombinant Proteins; D014758:Viral Core Proteins; D014759:Viral Envelope Proteins; D015678:Viral Structural Proteins; C107842:protein E1, Classical swine fever virus; C091015:glycoprotein E2, Hepatitis C virus