scholarly journals Identification of Epstein–Barr virus-infected CD27+ memory B-cells in liver or stem cell transplant patients

2011 ◽  
Vol 92 (11) ◽  
pp. 2590-2595 ◽  
Author(s):  
Yoshinori Ito ◽  
Shinji Kawabe ◽  
Seiji Kojima ◽  
Fumihiko Nakamura ◽  
Yukihiro Nishiyama ◽  
...  
Keyword(s):  
Liver Transplantation ◽  
Stem Cell ◽  
B Cells ◽  
Epstein Barr Virus ◽  
Peripheral Blood Lymphocytes ◽  
Haematopoietic Stem Cell ◽  
Cell Transplant ◽  
Barr Virus ◽  
Infected Cells ◽  
Epstein Barr

To analyse the phenotype of Epstein–Barr virus (EBV)-infected lymphocytes in EBV-associated infections, cells from eight haematopoietic stem cell/liver transplantation recipients with elevated EBV viral loads were examined by a novel quantitative assay designed to identify EBV-infected cells by using a flow cytometric detection of fluorescent in situ hybridization (FISH) assay. By this assay, 0.05–0.78 % of peripheral blood lymphocytes tested positive for EBV, and the EBV-infected cells were CD20+ B-cells in all eight patients. Of the CD20+ EBV-infected lymphocytes, 48–83 % of cells tested IgD positive and 49–100 % of cells tested CD27 positive. Additionally, the number of EBV-infected cells assayed by using FISH was significantly correlated with the EBV-DNA load, as determined by real-time PCR (r 2 = 0.88, P<0.0001). The FISH assay enabled us to characterize EBV-infected cells and perform a quantitative analysis in patients with EBV infection after stem cell/liver transplantation.

scholarly journals Cells Expressing the Epstein-Barr Virus Growth Program Are Present in and Restricted to the Naive B-Cell Subset of Healthy Tonsils

2000 ◽  
Vol 74 (21) ◽  
pp. 9964-9971 ◽  
Author(s):  
Alexandra M. Joseph ◽  
Gregory J. Babcock ◽  
David A. Thorley-Lawson
Keyword(s):  
B Cells ◽  
B Cell ◽  
Epstein Barr Virus ◽  
Cell Subset ◽  
Pcr Analysis ◽  
Virus Growth ◽  
Barr Virus ◽  
Latently Infected ◽  
Infected Cells ◽  
Epstein Barr

ABSTRACT In this paper we demonstrate, for the first time, that Epstein-Barr virus (EBV)-infected cells expressing the lymphoblastoid growth program are present in healthy carriers of the virus. Previously we observed that latently infected naive B cells are present in tonsils only when viral replication is detected, suggesting that these may represent newly infected B cells. We have tested this idea by performing a reverse transcription-PCR analysis for the expression of latent genes (EBNA2 and the EBNA3s) that are characteristically expressed only by newly infected cells expressing the growth latency program. EBNA2 expression is regularly detected in purified naive (IgD+) tonsillar B cells (13 of 16 tonsils tested) but was never found in the IgD− population (0 of 16). More detailed analysis revealed that the mRNAs for the latent genes EBNA1 (3 of 3 tonsils tested), EBNA3a (3 of 5), EBNA3b (3 of 5), EBNA3c (3 of 5), LMP1 (6 of 6), and LMP2 (5 of 6) were also present in the IgD+ population, but the EBNA1Q-K transcript, characteristic of nonlymphoblastoid forms of latency, was never detected (0 of 6). Finally, we demonstrate that the latently infected naive (IgD+) cells express CD80 (B7.1), a marker characteristically expressed on activated naive lymphoblasts but absent from resting naive B cells. The infected naive (IgD+) population in the tonsil therefore has the viral and cellular phenotype of a B-cell directly infected with EBV—an activated lymphoblast expressing the growth program.

scholarly journals Epstein-Barr Virus–Infected Resting Memory B Cells, Not Proliferating Lymphoblasts, Accumulate in the Peripheral Blood of Immunosuppressed Patients

1999 ◽  
Vol 190 (4) ◽  
pp. 567-576 ◽  
Author(s):  
Gregory J. Babcock ◽  
Lisa L. Decker ◽  
Richard B. Freeman ◽  
David A. Thorley-Lawson
Keyword(s):  
B Cells ◽  
Peripheral Blood ◽  
Epstein Barr Virus ◽  
Memory B Cells ◽  
Barr Virus ◽  
Latently Infected ◽  
Immunosuppressed Patients ◽  
Infected Cells ◽  
Epstein Barr ◽  
Healthy Carriers

When Epstein-Barr virus (EBV) infects B cells in vitro, the result is a proliferating lymphoblast that expresses at least nine latent proteins. It is generally believed that these cells are rigorously controlled in vivo by cytotoxic T cells. Consistent with this, the latently infected cells in the peripheral blood of healthy carriers are not lymphoblasts. Rather, they are resting memory B cells that are probably not subject to direct immunosurveillance by cytotoxic T lymphocytes (CTLs). When patients become immunosuppressed, the viral load increases in the peripheral blood. The expansion of proliferating lymphoblasts due to the suppressed CTL response is believed to account for this increase and is considered to be a major risk factor for posttransplant lymphoproliferative disease (PTLD) and AIDS-associated B cell lymphoma. Here we show that there is an increase in the numbers of latently infected cells in the peripheral blood of immunosuppressed patients. However, the cells are not proliferating lymphoblasts. They are all latently infected, resting, memory B cells—the same population of infected cells found in the blood of healthy carriers. These results are discussed in the context of a model for EBV persistence that explains why PTLD is usually limited to the lymph nodes.

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