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Epstein–Barr virus-induced B-cell transformation: quantitating events from virus binding to cell outgrowth

Authors :
Henri Jacques Delecluse
Claire Shannon-Lowe
Gouri Baldwin
Andrew I. Bell
Regina Feederle
Alan B. Rickinson
Source :
Journal of General Virology. 86:3009-3019
Publication Year :
2005
Publisher :
Microbiology Society, 2005.

Abstract

Epstein–Barr virus (EBV) infection and growth activation of human B cells is central to virus biology and disease pathogenesis, but is poorly understood in quantitative terms. Here, using virus at defined m.o.i., the different stages of this process at the single-cell level are followedin vitro. Virus binding to the B-cell surface, assayed by quantitative PCR, is highly efficient, particularly at the low m.o.i. values that most likely reflect physiologic eventsin vivo. However, only 10–15 % of bound virus genomes reach the cell nucleus, as visualized by sensitive fluorescencein situhybridization (FISH) assay; viral genomes acquired per cell nucleus range from 1 to >10, depending on the m.o.i. Thereafter, despite differences in initial genome load, almost all nuclear genome-positive cells then go on to express the virus-encoded nuclear antigen EBNA2, upregulate the cell activation antigen CD23 and transit the cell cycle. EBNA2-positive cells in the first cycle post-infection then grow out to lymphoblastoid cell lines (LCLs) just as efficiently as do cells limiting-diluted from already established LCLs. This study therefore identifies EBV genome delivery to the nucleus as a key rate-limiting step in B-cell transformation, and highlights the remarkable efficiency with which a single virus genome, having reached the nucleus, then drives the transformation programme.

Details

ISSN :
14652099 and 00221317
Volume :
86
Database :
OpenAIRE
Journal :
Journal of General Virology
Accession number :
edsair.doi.dedup.....28d5227be430ecf250fb1b650573bc53
Full Text :
https://doi.org/10.1099/vir.0.81153-0