In vitro detection of bovine immunodeficiency-like virus using monoclonal antibodies generated to a recombinant gag fusion protein.
An Escherichia coli recombinant fusion protein containing the major core protein of bovine immunodeficiency-like virus (BIV) was used to immunize mice for generation of monoclonal antibodies to BIV p26. Eight hybridomas specific for BIV p26 were identified and two antibodies, designated 104 and 142, were further characterized. Both 104 and 142 antibodies were isotyped as IgG1; they reacted specifically with both BIV p26 and the recombinant fusion protein in Western immunoblot analyses. However, the epitope specificity of the antibodies was different. Immunoperoxidase assays were used to determine if antibodies 104 and/or 142 could detect BIV replication in cell culture. Both antibodies were found to react with BIV-induced syncytia and individual BIV-infected cells. The antibodies were also used successfully in a focal immunoassay for quantitation of BIV-infected cells. These antibodies will provide valuable reagents for detection and quantitation of BIV replication in studies of viral pathogenesis and immunity.
Wannemuehler
Y
Y
Department of Microbiology, Immunology, and Preventive Medicine, College of Veterinary Medicine, Iowa State University, Ames 50011.
Isaacson
J
J
Wannemuehler
M
M
Wood
C
C
Roth
J A
JA
Carpenter
S
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000914:Antibodies, Viral / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D000956:Antigens, Viral / Q000276:immunology*; D002417:Cattle; D002459:Cell Fusion; D002460:Cell Line; D003588:Cytopathogenic Effect, Viral; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli; D005260:Female; D015683:Gene Products, gag / Q000276:immunology*; D007114:Immunization; D007118:Immunoassay; D016087:Immunodeficiency Virus, Bovine / Q000276:immunology / Q000302:isolation & purification* / Q000502:physiology; D007074:Immunoglobulin G / Q000276:immunology / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D014779:Virus Replication
D000911:Antibodies, Monoclonal; D000914:Antibodies, Viral; D000956:Antigens, Viral; D000939:Epitopes; D015683:Gene Products, gag; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins