Recombinant vaccinia viruses expressing hepatitis A virus structural polypeptides: detection of an anti-VP0 response in convalescent-phase sera.
We have generated a number of recombinant vaccinia viruses which expressed the hepatitis A virus (HAV) structural polypeptides VP1, VP2, VP3, and VP4, either alone or in combination. The relevant sequences encoding these polypeptides were amplified from cloned cDNA by PCR and then cloned into the insertion vector pGS62. The presence of the HAV structural polypeptide-encoding sequences in the recombinant viruses was confirmed by Southern blot analysis, whilst their transcription and translation were demonstrated by Northern (RNA) blot analysis and immunodetection, respectively. Immunoprecipitation studies using these constructs have detected the presence of an anti-VP0 response in human convalescent sera following HAV infection. The significance of this finding and the usefulness of these constructs in studying cell-mediated immunity during recovery from HAV infection are discussed.
Karayiannis
P
P
Department of Medicine, St. Mary's Hospital Medical School, Imperial College of Science, Technology and Medicine, London, United Kingdom. P.Karayiannis@sm.ic.ac.uk
O'Rourke
S
S
Watts
R
R
Waters
J
J
Hill
V
V
Carman
W F
WF
Thomas
H C
HC
D000914:Antibodies, Viral / Q000097:blood / Q000276:immunology; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D015153:Blotting, Western; D004279:DNA, Viral / Q000032:analysis; D015967:Gene Expression Regulation, Viral / Q000276:immunology; D005814:Genes, Viral / Q000235:genetics; D006506:Hepatitis A / Q000235:genetics / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000032:analysis; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D014616:Vaccinia virus / Q000235:genetics / Q000276:immunology*; D014764:Viral Proteins / Q000032:analysis; D015678:Viral Structural Proteins / Q000235:genetics
D000914:Antibodies, Viral; D004279:DNA, Viral; D012367:RNA, Viral; D011994:Recombinant Proteins; D014764:Viral Proteins; D015678:Viral Structural Proteins