Journal
JOURNAL OF VIROLOGICAL METHODS
Volume 217, Issue -, Pages 79-86Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2015.02.022
Keywords
Kaposi's sarcoma; Kaposi's sarcoma-associated herpesvirus; KSHV; human herpesvirus 8; BrK.219; high-titre virus stocks
Funding
- Collaborative Research Centre 900 of the Deutsche Forschungsgemeinschaft
- European Union Integrated Project INCA [LSHC-CT-2005-018704]
- DZIF (German Centre for Infection Research)
Ask authors/readers for more resources
Kaposi's sarcoma-associated herpesvirus (KSHV) is a gamma-2-Iymphotropic human oncogenic herpesvirus associated with Kaposi's sarcoma (KS) and two B-cell lymphoproliferative diseases, primary effusion lymphoma (PEL) and multicentric Castleman's disease (MCD). KSHV establishes latency soon after infection in vivo and in vitro. Consequently, it is technically difficult to generate high-titre virus stocks required for infection experiments in tissue culture. Currently used methods of KSHV stock production involve induction of the lytic/productive cycle in PEL cell lines or in adherent cell lines harbouring recombinant KSHV genomes. In this study, the BJAB-derived B-cell line BrK.219, which is infected latently with a recombinant KSHV (rKSHV.219), is used to produce high-titre virus stocks. BrK.219 cells enter the lytic KSHV replication cycle upon cross-linking of B-cell receptors (BCRs) with anti-IgM antibodies without the need for additional, potentially toxic chemical inducers. High cell concentrations can be cultured and induced easily in spinner flasks, saving time and resources. The established protocol allows the generation of KSHV virus stocks with titres of up to 106 IU/ml in unconcentrated culture supernatants, representing a 103-104-fold improvement compared to conventional methods. (C) 2015 Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available