Biological and Immunological Analyses of A Human Herpesvirus 8 Specific Glycoprotein.

• Project/Area Number: 11670308
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Virology
• Research Institution: Hyogo College of Medicine
• Principal Investigator: OKUNO Toshiomi Hyogo College of Medicine Faculty of Medicine Associate Professor, 医学部, 助教授 (10221152)
• Project Period (FY): 1999 – 2000
• Project Status: Completed (Fiscal Year 2000)
• Budget Amount: ¥3,100,000 (Direct Cost: ¥3,100,000); Fiscal Year 2000: ¥700,000 (Direct Cost: ¥700,000); Fiscal Year 1999: ¥2,400,000 (Direct Cost: ¥2,400,000)
• Keywords: human herpesvirus 8 / ADCC / ORFK8.1 / monoclonal antibody / glycoprotein / permissive cell line

Research Abstract

We established hybridoma clones secreting monoclonal antibodies against human herpesvirus 8 (HHV8) specific proteins. One of them, B332, recognized the protein encoded by open reading frame (ORF) K8.1, that the molecular weight was 38 kilo-dalton. It was synthesized at 48 hours after TPA treatment of HHV8 infected BC-3 cells, and localized on both cytoplasm and nucleus. HHV-8 could not infected and replicated in cells such as MT4, P3X63Ag8.653, Sp2/0-Ag14, HL-60, U937, THP-1, Molt3, Molt4, HSB2, Ramos, H9, TaY, LCL-TAS, B95-8, Raji, K562, D-2A, SHM-D33, HPB-ALL, HUT78, MDCK, U373MG, Hep2, B95a, HEL, CV-1, A549, Vero, 293, 293T, Balb/3T3, L929, 104, NBL-7, P3U1, PAI, MRC-5, SV1TG1, VSMC, BHK-21 and COS-1.
The ORFK8.1 gene was transfected into CV-1 cells, and stable cell lines expressing the protein were established. Using this cell line as a target for antibody dependent cell-mediated cytotoxicity assay, cytotoxic activity was observed in a patient's serum with Kaposi's sarcoma.