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. 2000 Apr;74(8):3478-85.
doi: 10.1128/jvi.74.8.3478-3485.2000.

Identification of antigenic proteins encoded by human herpesvirus 8 and seroprevalence in the general population and among patients with and without Kaposi's sarcoma

H Katano  1 , T Iwasaki, N Baba, M Terai, S Mori, A Iwamoto, T Kurata, T Sata
Affiliations

Identification of antigenic proteins encoded by human herpesvirus 8 and seroprevalence in the general population and among patients with and without Kaposi's sarcoma

H Katano et al. J Virol. 2000 Apr.

Abstract

To establish a sensitive and specific antibody assay, potent antigenic proteins encoded by human herpesvirus 8 (HHV8) were studied. Fifteen recombinant HHV8-encoded proteins were produced as glutathione S-transferase fusion proteins. The sera from AIDS-associated Kaposi's sarcoma (KS) patients reacted with four proteins encoded by open reading frames (ORFs) K8.1, 59, 65, and 73 in a Western blot assay. An enzyme-linked immunosorbent assay (ELISA) using these four proteins as antigens (mixed-antigen ELISA) revealed that all 26 sera derived from KS patients (24 with and 2 without human immunodeficiency virus infection) became positive for anti-HHV8 antibodies. The presence of HHV8 was demonstrated in 14 (1. 4%) of 1,004 sera from the Japanese general population and 10 (1.9%) of 527 sera from patients without HHV8-associated diseases. The presence of immunoglobulin G (IgG) and IgM antibodies against HHV8 examined further by the mixed-antigen ELISA and Western blotting revealed IgG antibody in all ELISA-positive sera, while IgM antibody against ORF K8.1 was absent. These data suggest that the ORF 73 and 65 proteins are potent antigens for a sensitive serological assay.

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Figures

FIG. 1
FIG. 1
Western blot analysis using TPA-induced TY-1 cell lysates revealed that the sera of AIDS-KS or PEL patients reacted with many bands and that the reaction patterns varied depending on the individual sera (lanes 1 to 6). No band was detected in the lanes containing the sera from a patient with infectious mononucleosis (IM) and a healthy blood donor (lanes 7 and 8). The reported sizes of the LANA and ORF59 bands are indicated on the right in kilodaltons.
FIG. 2
FIG. 2
Western blot analysis. Some of the recombinant GST fusion proteins were recognized by AIDS-KS or PEL patients' sera. The sera in lanes 1 to 8 are the same as those in Fig. 1. Only the recombinant protein bands in the membrane, whose sizes are listed in Table 1, are shown. IFA results are indicated at the bottom. The K8.1, ORF59, ORF65, and ORF73 proteins were recognized by some sera. Reactions with other proteins were weak. Lane 9 indicates that each protein was present on the membrane. IM, infectious mononucleosis.
FIG. 3
FIG. 3
Group scatter diagrams of ELISAs using the recombinant proteins. The scatter diagrams show reactions with various recombinant proteins in each ELISA. Compared with normal sera (right), AIDS-KS sera (left) reacted with the K8.1, ORF59, ORF65, ORF73N, ORF73C, and mixed proteins in ELISAs. The vertical axis indicates Ab, and the means and SD are shown on the right.
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