A model system for studying Kaposi’s sarcoma-associated herpesvirus (HHV-8) infection of B cells.

Tuesday, October 09, 2012 — Poster Session I
1:00 p.m. – 3:00 p.m	Natcher Conference Center, Building 45	NIAID	VIROL-4

* FARE Award Winner

Authors

• S.J. Dollery
• S. Moir
• E.A. Berger

Abstract

Kaposi’s sarcoma-associated herpesvirus (KSHV/HHV-8) is an HIV-associated gamma-herpesvirus. KSHV is causatively linked to the B-cell lymphoproliferative disorders multicentric Castleman’s disease and primary effusion lymphoma. Latently infected B cells are believed to be a major KSHV “reservoir”, and the virus has been shown replicate in human tonsillar B cells. Paradoxically, although several "irrelevant" cell lines (non-B cell, non-human) are permissive for in vitro KSHV infection, human B-cell lines and primary B cells are notoriously refractory to infection. We performed a strategic search of EBV-negative B lymphoma lines with the aim of identifying KSHV-susceptible B-cell lines. Using a recombinant KSHV encoding EGFP under a constitutive promoter and RFP under a lytic promoter (J. Vieira), plus a puromycin-resistance marker, we identified a novel B-cell line that was highly susceptible to KSHV infection (EGFP). Puromycin selection yielded homogenously infected cultures. Following puromycin removal, low-level latent infection was maintained. Various agents induced low-level lytic replication. These data indicate that KSHV can undergo latent and lytic replication in this cell line. Phenotypic analysis revealed clues about the origin of the cell line. These findings provide a first step in the establishment of a human B-cell line model to study the biologically relevant steps of KSHV infection.

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