Hepatitis B virus (HBV) X gene expression in human cells and anti-HBx antibodies detection in chronic HBV infection.
All mammalian hepatitis B virus genomes contain an open reading frame X (X-ORF) of unknown function which could encode a protein of 17 kDa. Using a plasmid containing the entire X-ORF preceded by the adenovirus type 2 major late promoter and its tripartite leader sequence efficient expression of the HBV X-gene was achieved. The X protein of 17 kDa was characterized by immunoblotting and immunoprecipitated with an antiserum prepared against a X fusion protein produced in E. coli. By cell fractionation and indirect immunofluorescence the X-protein was found at least in part associated with nuclei. Human cell extracts containing the X protein have been used to screen human sera for anti-HBx antibodies. Such antibodies were detected in sera from patients with active chronic hepatitis with ongoing viral replication. The efficient expression of the HBV X protein obtained will facilitate its functional analysis.
ER272-IRSC, Villejuif, France.
M
M
Levrero
O
O
Jean-Jean
C
C
Balsano
H
H
Will
M
M
Perricaudet
D002458:Cell Fractionation; D002460:Cell Line; D002908:Chronic Disease; D005455:Fluorescent Antibody Technique; D015967:Gene Expression Regulation, Viral; D005822:Genetic Vectors; D006509:Hepatitis B / Q000276:immunology*; D006510:Hepatitis B Antibodies / Q000032:analysis*; D006515:Hepatitis B virus / Q000235:genetics* / Q000276:immunology; D006801:Humans; D015151:Immunoblotting; D010957:Plasmids; D011233:Precipitin Tests; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D014162:Transfection; D054334:Viral Regulatory and Accessory Proteins
D006510:Hepatitis B Antibodies; D015534:Trans-Activators; D054334:Viral Regulatory and Accessory Proteins; C057628:hepatitis B virus X protein