Determination of Epstein-Barr virus (EBV) load by RT-PCR and cellular dilution

1998 ◽  
Vol 12 (6) ◽  
pp. 427-430 ◽  
Author(s):  
R.J. Orentas
Keyword(s):  
Epstein Barr Virus ◽  
Rt Pcr ◽  
Barr Virus ◽  
Epstein Barr

Modulation of Caspase-8 and FLICE-Inhibitory Protein Expression as a Potential Mechanism of Epstein-Barr Virus Tumorigenesis in Burkitt’s Lymphoma

Blood ◽  
1999 ◽  
Vol 94 (5) ◽  
pp. 1727-1737 ◽  
Author(s):  
Clifford G. Tepper ◽  
Michael F. Seldin
Keyword(s):  
Cell Lines ◽  
Epstein Barr Virus ◽  
Burkitt’S Lymphoma ◽  
Burkitt's Lymphoma ◽  
Pcr Analysis ◽  
Caspase 8 ◽  
Rt Pcr ◽  
Inhibitory Protein ◽  
Barr Virus ◽  
Epstein Barr

Abstract Ligation of the Fas receptor induces death-inducing signaling complex (DISC) formation, caspase activation, and subsequent apoptotic death of several cell types. Epstein-Barr virus (EBV)-positive group III Burkitt’s lymphoma (BL) cell lines have a marked resistance to Fas-mediated apoptosis, although expressing each of the DISC components, Fas/ APO-1–associated death domain protein (FADD), and caspase-8 (FLICE/MACH/Mch5). The apoptotic pathway distal to the DISC is intact because ceramide analogs, staurosporine, and granzyme B activate caspase-3 and induce apoptosis. Fas resistance was not explained by the putative death-attenuating caspase-8 isoforms. However, while Fas-activated cytosolic extracts from sensitive cells were capable of processing both procaspase-8 and procaspase-3 into active subunit forms, resistant cell extracts did not possess either of these activities. Accordingly, reverse transcriptase-polymerase chain reaction (RT-PCR) analysis showed higher transcript levels for the FLICE-inhibitory protein (FLIPL) in resistant cells and the ratio of caspase-8 to FLIPLmeasured by competition RT-PCR analysis directly correlated with susceptibility to Fas-mediated apoptosis of all cell lines. In addition, modification of the caspase-8/FLIPL ratio by caspase-8 or FLIPL overexpression was able to alter the susceptibility status of the cell lines tested. Our results imply that the relative levels of caspase-8 and FLIPL are an important determinant of susceptibility to Fas-mediated apoptosis.

scholarly journals Epstein-Barr Virus-Induced Changes in B-Lymphocyte Gene Expression

2002 ◽  
Vol 76 (20) ◽  
pp. 10427-10436 ◽  
Author(s):  
Kara L. Carter ◽  
Ellen Cahir-McFarland ◽  
Elliott Kieff
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
B Lymphocytes ◽  
Epstein Barr Virus ◽  
Molecular Mechanisms ◽  
B Lymphocyte ◽  
Protein Biosynthesis ◽  
Rt Pcr ◽  
Barr Virus ◽  
Epstein Barr

ABSTRACT To elucidate the mechanisms by which Epstein-Barr virus (EBV) latency III gene expression transforms primary B lymphocytes to lymphoblastoid cell lines (LCLs), the associated alterations in cell gene expression were assessed by using 4,146 cellular cDNAs arrayed on nitrocellulose filters and real-time reverse transcription-PCR (RT-PCR). A total of 1,405 of the 4,146 cDNAs were detected using cDNA probes from poly(A)+ RNA of IB4 LCLs, a non-EBV-infected Burkitt's lymphoma (BL) cell line, BL41, or EBV latency III-converted BL41 cells (BL41EBV). Thirty-eight RNAs were consistently twofold more abundant in the IB4 LCL and BL41EBV than in BL41 by microarray analysis. Ten of these are known to be EBV induced. A total of 23 of 28 newly identified EBV-induced genes were confirmed by real-time RT-PCR. In addition, nine newly identified genes and CD10 were EBV repressed. These EBV-regulated genes encode proteins involved in signal transduction, transcription, protein biosynthesis and degradation, and cell motility, shape, or adhesion. Seven of seven newly identified EBV-induced RNAs were more abundant in newly established LCLs than in resting B lymphocytes. Surveys of eight promoters of newly identified genes implicate NF-κB or PU.1 as potentially important mediators of EBV-induced effects through LMP1 or EBNA2, respectively. Thus, examination of the transcriptional effects of EBV infection can elucidate the molecular mechanisms by which EBV latency III alters B lymphocytes.

scholarly journals Comparison of Three Automated Immunoassay Methods for the Determination of Epstein-Barr Virus-Specific Immunoglobulin M

2010 ◽  
Vol 17 (4) ◽  
pp. 559-563 ◽  
Author(s):  
Mario Berth ◽  
Eugene Bosmans
Keyword(s):  
Epstein Barr Virus ◽  
Antibody Test ◽  
Immunoglobulin M ◽  
Nuclear Antigen ◽  
Two Phase ◽  
Barr Virus ◽  
Epstein Barr ◽  
Automated Immunoassay ◽  
A Prospective Study

ABSTRACT In this study we compared the performances of three commercially available Epstein-Barr virus (EBV) immunoglobulin M (IgM) assays on highly automated immunoassay platforms: BioPlex 2200 (Bio-Rad Laboratories), Immulite 2000 (Siemens Healthcare Diagnostics), and Liaison (DiaSorin). As a confirmatory method, immunoblotting was performed. The specificity of the three EBV IgM assays was evaluated by testing 293 selected sera from patients with various infectious and noninfectious diseases. After the exclusion of 30 samples, the specificities were 96.2% for Liaison, 98.1% for Immulite, and 97.0% for BioPlex. For evaluation of the sensitivity, samples from 70 consecutive patients with a positive heterophile antibody test were examined, irrespective of clinical or biological findings. After the exclusion of six samples, the sensitivities were 89.1% for Liaison, 84.4% for Immulite, and 89.1% for BioPlex. Finally, in a prospective study performed with 500 samples obtained from consecutive patients and sent in by general practitioners, we also determined Epstein-Barr nuclear antigen IgG and viral capsid antigen IgG in a two-phase approach. Concordance of the EBV serologic status was 96.2% between Liaison and Immulite, 96.4% between Immulite and BioPlex, and 97.8% between BioPlex and Liaison. The three EBV IgM immunoassays that we evaluated have acceptable and comparable performances.

scholarly journals Cloning of the Rhesus Lymphocryptovirus Viral Capsid Antigen and Epstein-Barr Virus-Encoded Small RNA Homologues and Use in Diagnosis of Acute and Persistent Infections

2000 ◽  
Vol 38 (9) ◽  
pp. 3219-3225 ◽  
Author(s):  
Pasupuleti Rao ◽  
Hua Jiang ◽  
Fred Wang
Keyword(s):  
Small Rna ◽  
Epstein Barr Virus ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Viral Capsid ◽  
Viral Capsid Antigen ◽  
Rt Pcr ◽  
Barr Virus ◽  
Peptide Elisa ◽  
Epstein Barr

Epstein-Barr virus (EBV) is the most common cause of infectious mononucleosis and is associated with the development of several human malignancies. A closely related herpesvirus in the same lymphocryptovirus (LCV) genera as EBV naturally infects rhesus monkeys and provides an important animal model for studying EBV pathogenesis. We cloned the small viral capsid antigen (sVCA) homologue from the rhesus LCV and developed a peptide enzyme-linked immunosorbent assay (ELISA) to determine whether epitopes in the rhesus LCV sVCA are a reliable indicator of rhesus LCV infection. In order to define a “gold standard” for rhesus LCV infection, we also cloned the EBV-encoded small RNA 1 (EBER1) and EBER2 homologues from rhesus LCV and developed a reverse transcription (RT)-PCR assay to detect persistent LCV infection in rhesus monkey peripheral blood lymphocytes. Animals from a conventional and a hand-reared colony were studied to compare the prevalence of rhesus LCV infection in the two groups. There was a 100% correlation between the peptide ELISA and EBER RT-PCR results for rhesus LCV infection. In addition, specificity for LCV infection and exclusion of potential cross-reactivity to the rhesus rhadinovirus sVCA homologue could be demonstrated using sera from experimentally infected animals. These studies establish two novel assays for reliable diagnosis of acute and persistent rhesus LCV infections. The rhesus LCV sVCA peptide ELISA provides a sensitive and reliable assay for routine screening, and these studies of the hand-reared colony confirm the feasibility of raising rhesus LCV-naive animals.

Comparison of Abbott Architect ® , Siemens Immulite ® , and Diasorin Liaison ® for determination of Epstein–Barr virus serological diagnosis

2018 ◽  
Vol 90 (2) ◽  
pp. 96-101 ◽  
Author(s):  
Catherine François ◽  
Christine Segard ◽  
Maryline Bouvier ◽  
Martine Stefanski ◽  
Christine Pannier ◽  
...  
Keyword(s):  
Epstein Barr Virus ◽  
Serological Diagnosis ◽  
Barr Virus ◽  
Epstein Barr ◽  
Abbott Architect

Chromosome banding, isoenzyme studies and determination of epstein-barr virus DNA content on human burkitt lymphoma/mouse hybrids

1977 ◽  
Vol 20 (6) ◽  
pp. 849-853 ◽  
Author(s):  
Jack Spira ◽  
Susanne Povey ◽  
Francis Wiener ◽  
George Klein ◽  
Maria Andersson-Anvret
Keyword(s):  
Dna Content ◽  
Burkitt Lymphoma ◽  
Epstein Barr Virus ◽  
Chromosome Banding ◽  
Barr Virus ◽  
Epstein Barr
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