Identification, expression, and immunogenicity of Kaposi's sarcoma-associated herpesvirus-encoded small viral capsid antigen.
We describe a recombinant antigen for use in serologic tests for antibodies to Kaposi's sarcoma (KS)-associated herpesvirus (KSHV). The cDNA for a small viral capsid antigen (sVCA) was identified by immunoscreening of a library prepared from the BC-1 body cavity lymphoma cell line induced into KSHV lytic gene expression by sodium butyrate. The cDNA specified a 170-amino-acid peptide with homology to small viral capsid proteins encoded by the BFRF3 gene of Epstein-Barr virus and the ORF65 gene of herpesvirus saimiri. KSHV sVCA was expressed from a 0.85-kb mRNA present late in lytic KSHV replication in BC-1 cells. This transcript was sensitive to phosphonoacetic acid and phosphonoformic acid, inhibitors of herpesvirus DNA replication. KSHV sVCA expressed in mammalian cells or Escherichia coli or translated in vitro was recognized as an antigen by antisera from KS patients. Rabbit antisera raised to KSHV sVCA expressed in E. coli detected a 22-kDa protein in KSHV-infected human B cells. Overexpressed KSHV sVCA purified from E. coli and used as an antigen in immunoblot screening assay did not cross-react with EBV BFRF3. Antibodies to sVCA were present in 89% of 47 human immunodeficiency virus (HIV)-positive patients with KS, in 20% of 54 HIV-positive patients without KS, but in none of 122 other patients including children born to HIV-seropositive mothers and patients with hemophilia, autoimmune disease, or nasopharyngeal carcinoma. Low-titer antibody was detected in three sera from 28 healthy subjects. Antibodies to recombinant sVCA correlate with KS in high-risk populations. Recombinant sVCA can be used to examine the seroepidemiology of infection with KSHV in the general population.
Lin
S F
SF
Department of Molecular Biophysics & Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06520, USA.
Sun
R
R
Heston
L
L
Gradoville
L
L
Shedd
D
D
Haglund
K
K
Rigsby
M
M
Miller
G
G
D017088:AIDS-Related Opportunistic Infections / Q000097:blood / Q000276:immunology / Q000821:virology*; D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D019556:COS Cells; D002213:Capsid / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins; D002460:Cell Line; D003001:Cloning, Molecular; D003429:Cross Reactions; D004279:DNA, Viral; D004926:Escherichia coli; D015870:Gene Expression; D019288:Herpesvirus 8, Human / Q000235:genetics / Q000276:immunology*; D006801:Humans; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000032:analysis; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012514:Sarcoma, Kaposi / Q000097:blood / Q000276:immunology / Q000821:virology*; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured; D014764:Viral Proteins
D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D004279:DNA, Viral; D012367:RNA, Viral; D011993:Recombinant Fusion Proteins; C041788:Epstein-Barr viral capsid antigen; C119019:ORF65 protein, human herpesvirus 8; D014764:Viral Proteins