Baculovirus expression of the glycoprotein gene of Lassa virus and characterization of the recombinant protein.
A recombinant baculovirus was constructed that expresses the glycoprotein gene of Lassa virus (Josiah strain) under the transcriptional control of the polyhedrin promoter. The expressed protein (B-LSGPC) comigrated with the authentic viral glycoprotein as observed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), was reactive with monoclonal antibodies (MAbs) in Western blots, and was glycosylated. Although the recombinant protein was not processed into the mature glycoproteins, G1 and G2, it demonstrated reactivity with all known epitopes as measured by indirect immunofluorescence (IFA), and it was immunogenic, eliciting antisera in rabbits that recognized whole virus in IFAs. Regarding future applications to diagnostic assays, the recombinant glycoprotein proved to be an effective substitute for Lassa virus-infected mammalian cells in IFAs and it was able to distinguish sera from several human cases of Lassa fever, against a panel of known negative sera of African origin, in an enzyme immunoassay (EIA).
Hummel
K B
KB
Division of Viral and Rickettsial Diseases, Centers for Disease Control, Atlanta, GA 30333.
Martin
M L
ML
Auperin
D D
DD
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics; D016367:Baculoviridae / Q000235:genetics*; D002460:Cell Line; D015870:Gene Expression; D005814:Genes, Viral; D006023:Glycoproteins / Q000235:genetics / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques; D007835:Lassa Fever / Q000175:diagnosis; D007836:Lassa virus / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D014764:Viral Proteins / Q000235:genetics* / Q000276:immunology
D000956:Antigens, Viral; D006023:Glycoproteins; D011994:Recombinant Proteins; D014764:Viral Proteins