Comparison of the immunoglobulin-G-specific seroreactivity of different recombinant antigens of the human herpesvirus 8.
The open reading frames ORF 52, ORF 65, K12, and K8.1 of the human herpesvirus 8 (HHV8) were expressed as glutathione-S-transferase (GST) fusion proteins and analysed by Western blotting (WB) and enzyme-linked immunosorbent assay (ELISA). The open reading frame (ORF) 65 and K8.1 antigens gave the highest reactivity (71%) in sera from HIV-dependent Kaposi's sarcoma (KS) patients. Therefore both antigens appear to be essential for HHV8 diagnostics, whereas ORF K12 and ORF 52 were of minor importance. Using polymerase chain reaction (PCR) out of the peripheral blood of these KS patients, 48% were detected as positive. By testing an N-terminal-deleted construct (amino acid 80-171) of ORF 65, we could show that the N-terminal region of this protein is essential to mediate full immunogenic reactivity. By analysing different deletion mutants of ORF K8.1, the major epitope was found to be located between aa 29 and 101. The prevalence of antibodies directed against the different antigens was determined for healthy blood donors to be 3-6%. The different antibody patterns obtained in HIV-patients with and without KS support the hypothesis that different antibody profiles develop during the course of KS.
Lang
D
D
Research Department, Biotest AG, Dreieich, D-63303, Germany.
Hinderer
W
W
Rothe
M
M
Sonneborn
H H
HH
Neipel
F
F
Raab
M
M
Rabenau
H
H
Masquelier
B
B
Fleury
H
H
D000956:Antigens, Viral / Q000276:immunology*; D015139:Blotting, Southern; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D005982:Glutathione Transferase / Q000235:genetics; D015658:HIV Infections / Q000821:virology; D019288:Herpesvirus 8, Human / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology*; D016366:Open Reading Frames; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000276:immunology
D000956:Antigens, Viral; D007074:Immunoglobulin G; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase