Characterization of a major protein with a molecular weight of 160,000 associated with the viral capsid of Epstein-Barr virus

Abstract

A monoclonal antibody designated V3 was produced against a late protein associated with the Epstein-Barr virus-induced viral capsid antigen complex. The antibody reacted with discrete patches in the nuclei of infected cells as well as with virus particles, as shown by immunofluorescence and ultrastructural immunoperoxidase staining. The molecular weight of the protein precipitated by this monoclonal antibody was ca. 160,000. All anti-viral capsid antigen antibody-positive sera tested in an enzyme-linked immunosorbent assay reacted with this purified protein. The synthesis of the antigen was inhibited by phosphonoacetic acid but was not affected by tunicamycin, indicating that this was a late nonglycosylated viral protein. No differences were noted between the protein isolated from the P3HR-1 and B-95-8 cell lines as determined by immunoprecipitation and peptide mapping. By isoelectric focusing, this protein had a pl on the basic side ranging from 7.5 to 9.0.

Identification and molecular characterization of two diagnostically relevant marker proteins of the Epstein‐Barr virus capsid antigen complex

“…IgM and IgA responses, temporarily detectable in the acute stage of IM, are absent in sera of healthy EBV carriers. In general, the dominant antibody response of healthy EBV carriers consists of IgG antibodies directed against EBNAl (72 kD) [Miller et al, 19871;the VCA proteins p18, p40, gp125, p160 [this study;Vroman et al, 1985;Luka et al, 19841; and the polypeptides p36/38 of the EA complex, representing the Zebra protein.…”

“…For diagnosis of IM and nasopharyngeal carcinoma (NPC), the 138 kD (BALFB) and the 47-54 kD (BMRF1) EA(D) molecules appear to be good candidates [Dolken et al, 1987;Motz et al, 1986;Middeldorp and Herbrink, 19881. Viral thymidine kinase (BXLF1) and DNase (BGLF5) may be used as more selective markers for NPC progression [Littler et al, 1991;Ooka et al, 19911. A few studies only have examined the molecular nature of diagnostic marker molecules of the VCA complex, which to date include the major capsid protein (BcLF1) and several membrane/envelope associated polypeptides [Vroman et al, 1985;Wolf et al, 1987;Luka et al, 19841. Even fewer studies have addressed 162 van Grunsven et al…”

Characterization of the restricted component of Epstein-Barr virus early antigens as a cytoplasmic filamentous protein

“…The mice were immunized four times with 25 ,ug of purified protein in the presence of 0.1% sodium dodecyl sulfate (SDS). Spleen cells were prepared 4 days after the last immunization, and the cells were fused to NS-1 cells by the method of Vroman et al (24). Supernatants from wells with visible growth were tested for antibodies by enzyme-linked immunosorbent assay (ELISA) with affinitypurified antigens as the target (10) and by immunofluorescence on virus-producing and nonproducer cell lines (18).…”

“…Immunofluorescence procedure. Acetone-or methanolfixed smears of activated cells were stained by immunofluorescence as previously described in detail (8,19,24) by using fluorescein-conjugated goat anti-mouse immunoglobulin G (IgG). The stained smears were mounted in 50% buffered glycerol and then examined with a fluorescence microscope (American Optical Corp., Buffalo, N.Y.).…”

Development of an enzyme‐linked immunosorbent assay (ELISA) for detecting IgA antibodies to the epstein‐barr virus

“…However, with the advent of MAb technology, this is now possible. A number of polypeptides associated with the different EBV antigen complexes have now been identified, purified and partially characterized (Hoffman et al, 1980;Thorley-Lawson and Geilinger, 1980;Qualtibre et al, 1982b;Pearson et al, 1983aPearson et al, , 1987Takada et al, 1983;Kishishita et al, 1984;Vroman et al, 1985). In addition, new assays have been developed to monitor both IgG and IgM antibodies to individual components of these complexes purified by immunoaffinity chromatography .…”