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Journal of Clinical Microbiology, July 2002, p. 2533-2544, Vol. 40, No. 7
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.7.2533-2544.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Detection of Epstein-Barr Virus Genomes in Peripheral Blood B Cells from Solid-Organ Transplant Recipients by Fluorescence In Situ Hybridization

Department of Infectious Diseases and Microbiology,¹ Department of Biostatistics, Graduate School of Public Health,⁴ Department of Cell Biology and Physiology, School of Medicine, University of Pittsburgh,⁵ Departments of Pediatrics,² Surgery, Children's Hospital of Pittsburgh, Pittsburgh, Pennsylvania 15213³

Received 13 November 2001/ Returned for modification 12 January 2002/ Accepted 14 March 2002

Resolution of Epstein-Barr Virus (EBV) infection in pediatric solid-organ transplant recipients often leads to an asymptomatic carrier state characterized by a persistently elevated circulating EBV load that is 2 to 4 orders of magnitude greater than the load typical of healthy latently infected individuals. Elevated EBV loads in immunosuppressed individuals are associated with an increased risk for development of posttransplant lymphoproliferative disease. We have performed fluorescence in situ hybridization (FISH) studies with peripheral blood B cells from carriers of persistent EBV loads in order to directly quantitate the number of EBV genomes per infected cell. Patients were assigned to two groups on the basis of the level of the persistent load (low-load carriers, 8 to 200 genomes/10⁵ peripheral blood lymphocytes; high-load carriers, >200 genomes/10⁵ peripheral blood lymphocytes). FISH analysis revealed that the low-load carriers predominantly had circulating virus-infected cells harboring one or two genome copies/cell. High-load carriers also had cells harboring one or two genome copies/cell; in addition, however, they carried a distinct population of cells with high numbers of viral genome copies. The increased viral loads correlated with an increase in the frequency of cells containing high numbers of viral genomes. We conclude that low-load carriers possess EBV-infected cells that are in a state similar to normal latency, whereas high-load carriers possess two populations of virus-positive B cells, one of which carries an increased number of viral genomes per cell and is not typical of normal latency.

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