Development of a fluorescence-based assay to screen antiviral drugs against Kaposi's sarcoma associated herpesvirus

Nun TK, Kroll DJ, Oberlies NH, Soejarto DD, Case RJ, Piskaut P, Matainaho T, Hilscher C, Wang L, Dittmer DP, Gao SJ, Damania B
Source: Mol Cancer Ther
Publication Date: (2007)
Issue: 6(8): 2360-70
Research Area:
Cancer Research/Cell Biology
Immunotherapy / Hematology
Cells used in publication:
Species: human
Tissue Origin: blood
Nucleofector® I/II/2b
To establish BJAB-KSHV cell line, the complete KSHV (Karposi`s sarcoma herpesvirus) genome introduced into a bacterial artificial chromosome (BAC) and nucleofected into BJAB cell line. The KSHV-BAC contained a mammalian hygromycin antibiotic resistance marker and a GFP expression cassette. Positive cells were selected for a minimum of two weeks.
Tumors associated with Kaposi's sarcoma-associated herpesvirus infection include Kaposi's sarcoma, primary effusion lymphoma, and multicentric Castleman's disease. Virtually all of the tumor cells in these cancers are latently infected and dependent on the virus for survival. Latent viral proteins maintain the viral genome and are required for tumorigenesis. Current prevention and treatment strategies are limited because they fail to specifically target the latent form of the virus, which can persist for the lifetime of the host. Thus, targeting latent viral proteins may prove to be an important therapeutic modality for existing tumors as well as in tumor prevention by reducing latent virus load. Here, we describe a novel fluorescence-based screening assay to monitor the maintenance of the Kaposi's sarcoma-associated herpesvirus genome in B lymphocyte cell lines and to identify compounds that induce its loss, resulting in tumor cell death.