The glycoprotein products of varicella-zoster virus gene 14 and their defective accumulation in a vaccine strain (Oka).
Many characteristics of the putative protein encoded by varicella-zoster virus (VZV) open reading fram (ORF) 14 indicate that it is a glycoprotein, which has been designated gpV. To identify the protein products of the gene, the coding sequences were placed under the control of the vaccinia virus p7.5 promoter and recombinant vaccinia viruses were constructed. Heterogeneous polypeptides with molecular weights of 95,000 to 105,000 (95K to 105K polypeptides) were expressed in cells infected by a vaccinia virus recombinant (vKIP5) containing ORF 14 from VZV Scott but were not expressed by control vaccinia viruses. These polypeptides were recognized by antibodies present in human sera that contained high levels of anti-VZV antibodies. Conversely, antisera raised in rabbits inoculated with vKIP5 reacted specifically with heterogeneous 95K to 105K polypeptides present in VZV Scott-infected but not uninfected cells; these polypeptides show a patchy plasma membrane fluorescence pattern in VZV Scott-infected cells. These same antisera neutralized VZV strain Scott infectivity in the absence of complement. Endoglycosidase F treatment of isolated gpV polypeptides and tunicamycin treatment of cells infected with the vKIP5 recombinant indicated that the polypeptides were glycosylated. Three sets of data imply that the VZV strain Oka, which has been used to produce a live attenuated virus vaccine, accumulates low levels of gpV polypeptides relative to wild-type strains: (i) blocking of antibodies in human sera with excess VZV Oka-infected cell antigen yielded residual antibodies which were reactive with the 95K to 105K gpV polypeptides expressed in cells infected by VZV strain Scott and by the vKIP5 vaccinia virus recombinant, but not with Oka-infected cell polypeptides; (ii) antisera raised to vKIP5 detected very low levels of reactive polypeptides made in VZV Oka-infected cells and neutralized VZV Oka virus much less efficiently than VZV Scott; and (iii) comparisons of the reactivity of sera from live attenuated virus vaccine vaccinees with sera derived from patients recovering from wild-type infections indicated greatly reduced levels of gpV-specific antibodies in some vaccinees.
Kinchington
P R
PR
Department of Biochemistry, Uniformed Services University of the Health Sciences, Bethesda, Maryland, 20814-4799.
Ling
P
P
Pensiero
M
M
Moss
B
B
Ruyechan
W T
WT
Hay
J
J
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D005455:Fluorescent Antibody Technique; D005814:Genes, Viral; D006023:Glycoproteins / Q000235:genetics*; D014645:Herpesvirus 3, Human / Q000235:genetics*; D006801:Humans; D007106:Immune Sera; D008969:Molecular Sequence Data; D009500:Neutralization Tests; D010957:Plasmids; D011401:Promoter Regions, Genetic; D015183:Restriction Mapping; D014616:Vaccinia virus / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics*
D004279:DNA, Viral; D006023:Glycoproteins; D007106:Immune Sera; D014764:Viral Proteins