A convenient semi-quantitative method for the diagnosis of Epstein-Barr virus reactivation

Venard, Véronique; Carret, Anne‐Sophie; Pascal, N.; Rihn, Bertrand; Bordigoni, Pierre; Faou, Alain Le

doi:10.1007/s007050070051

Cited by 10 publications

(13 citation statements)

References 14 publications

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“…By using our multiplex real-time PCR assay, we have already shown earlier that the cross-sectional detection rate of EBV DNA was 84.6% in PBMC of healthy blood donors (11). The detection rate for EBV DNA in a recent study was 28% for bone marrow recipients by means of nested semiquantitative PCR (28). Another study assessing Light Cycler-based PCR found a detectable viral load in 37.5% of healthy individuals (4).…”

Section: Discussionmentioning

confidence: 64%

Molecular Parameters for Precise Diagnosis of Asymptomatic Epstein-Barr Virus Reactivation in Healthy Carriers

Maurmann¹,

Fricke²,

Wagner³

et al. 2003

J Clin Microbiol

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Asymptomatic Epstein-Barr virus (EBV) reactivations periodically occur in oral mucosa-associated lymphoid tissues. Until now, EBV reactivation has been diagnosed by serologic profiles that suggest virus replication. Serologic responses, however, are delayed and do not necessarily indicate ongoing replicative activity. The aim of the present study was to establish in healthy carriers parameters for a molecular diagnosis of reactivated EBV infection. Recent studies emphasized the association of an increase in peripheral-B-cell viral load with replicative activity at remote sites. Therefore, real-time PCR was used to quantitate EBV genomes in the peripheral blood mononuclear cells (PBMC) (viral load) and plasma samples (viremia) of 22 healthy EBV-seropositive blood donors over a period of 15 months. Furthermore, transcription of the immediate-early gene encoding BZLF1 was investigated in the PBMC of all volunteers. Serology suggested reactivation in nine donors, of whom all but one showed at least once a significant increase in viral load. Another five individuals also exhibited significant changes in viral load but no serologic response. Of the 13 volunteers with significant increases in viral load, 6 had a period of viremia accompanying the rise in viral load. A stable viral load without viremia and negative serology was seen in eight adults. BZLF1 mRNA was undetectable throughout. We conclude that for healthy subjects serology underestimates the frequency of asymptomatic EBV reactivations. Prospective examination of peripheral viral load and viremia is suitable for the exact diagnosis of EBV reactivation, which might be of advantage for immunocompromised patients in whom EBV reactivations are considerably harmful.

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Section: Discussionmentioning

confidence: 64%

Molecular Parameters for Precise Diagnosis of Asymptomatic Epstein-Barr Virus Reactivation in Healthy Carriers

Maurmann¹,

Fricke²,

Wagner³

et al. 2003

J Clin Microbiol

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“…PCR to assess TCRg gene rearrangement and screen for oncogene incorporation was carried out as described previously (1,21,(25)(26)(27). Quantitative reverse transcriptase PCR (qRT-PCR) to measure gene expression was carried out as described previously (24).…”

Section: Pcr and Quantitative Reverse Transcriptase Pcrmentioning

confidence: 99%

Survival Signals and Targets for Therapy in Breast Implant–Associated ALK⁻Anaplastic Large Cell Lymphoma

Lechner¹,

Megiel²,

Church³

et al. 2012

Clin Cancer Res

110

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Purpose: Anaplastic lymphoma kinase (ALK)-negative, T-cell, anaplastic, non-Hodgkin lymphoma (T-ALCL) in patients with textured saline and silicone breast implants is a recently recognized clinical entity for which the etiology and optimal treatment remain unknown.Experimental Design: Using three newly established model cell lines from patient biopsy specimens, designated T-cell breast lymphoma (TLBR)-1 to -3, we characterized the phenotype and function of these tumors to identify mechanisms of cell survival and potential therapeutic targets.Results: Cytogenetics revealed chromosomal atypia with partial or complete trisomy and absence of the NPM-ALK (2;5) translocation. Phenotypic characterization showed strong positivity for CD30, CD71, T-cell CD2/5/7, and antigen presentation (HLA-DR, CD80, CD86) markers, and interleukin (IL)-2 (CD25, CD122) and IL-6 receptors. Studies of these model cell lines showed strong activation of STAT3 signaling, likely related to autocrine production of IL-6 and decreased SHP-1. STAT3 inhibition, directly or by recovery of SHP-1, and cyclophosphamide-Adriamycin-vincristine-prednisone (CHOP) chemotherapy reagents, effectively kill cells of all three TLBR models in vitro and may be pursued as therapies for patients with breast implant-associated T-ALCLs.Conclusions: The TLBR cell lines closely resemble the primary breast implant-associated lymphomas from which they were derived and as such provide valuable preclinical models to study their unique biology.

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“…If so, the mere presence of EBV in the blood should ensure a close follow-up of the patient in search of the first symptoms of PTLD. Viral load may be determined using circulating mononuclear cells [11,14,15,17,19], serum [17,20], plasma [19], or whole blood [6,8,12,16]. Although no consensus has been established, unfractionated whole blood seems to be the substrate of choice and is most commonly used [21].…”

Section: Discussionmentioning

confidence: 99%

“…Amplification products were analyzed on agarose gels containing ethidium bromide. The sensitivity of the reaction was of 200-2,000 copies, as determined using Namalwa cells [15]. Results were given as positive for the number of cells present in 5 l of the crude extract, positive for 500 or 100 cells when dilutions were tested.…”

Section: Viral Loadmentioning

confidence: 99%

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Long-term follow-up of Epstein-Barr virus viremia in pediatric recipients of renal transplants

et al. 2004

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The common observation of Epstein-Barr virus (EBV) viremia in pediatric recipients of renal transplants and the occurrence of an EBV-related pulmonary leiomyoma prompted us to intensify the follow-up of EBV infections from 1995 to October 2000. Follow-up included serology and detection of viral DNA in blood using a semi-quantitative nested polymerase chain reaction (PCR) and later a real-time PCR with higher sensitivity. The aim of this study was the early detection of primary infections or reactivations. We obtained 250 samples from 32 patients. EBV DNA detection was consistently negative in 14 patients. There were 5 patients that were considered at risk for post-transplant lymphoproliferative disease, as they were EBV seronegative and were given a kidney from a positive donor. Of these, 4 had at least one episode of high-level EBV viremia. During these episodes, an absence of noticeable symptoms that could be related to EBV was noted for all but 1 patient. This child presented with severe neutropenia 1 month after grafting and, 28 months later, several nodules of pulmonary leiomyoma, which were found to be EBV related. Four episodes of high-level viremia were observed before the discovery of the leiomyoma. Viral DNA detection is important for the follow-up of such patients that are especially at risk of serious complications of EBV infections.

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A convenient semi-quantitative method for the diagnosis of Epstein-Barr virus reactivation

Cited by 10 publications

References 14 publications

Molecular Parameters for Precise Diagnosis of Asymptomatic Epstein-Barr Virus Reactivation in Healthy Carriers

Molecular Parameters for Precise Diagnosis of Asymptomatic Epstein-Barr Virus Reactivation in Healthy Carriers

Survival Signals and Targets for Therapy in Breast Implant–Associated ALK⁻Anaplastic Large Cell Lymphoma

Long-term follow-up of Epstein-Barr virus viremia in pediatric recipients of renal transplants

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A convenient semi-quantitative method for the diagnosis of Epstein-Barr virus reactivation

Cited by 10 publications

References 14 publications

Molecular Parameters for Precise Diagnosis of Asymptomatic Epstein-Barr Virus Reactivation in Healthy Carriers

Molecular Parameters for Precise Diagnosis of Asymptomatic Epstein-Barr Virus Reactivation in Healthy Carriers

Survival Signals and Targets for Therapy in Breast Implant–Associated ALK−Anaplastic Large Cell Lymphoma

Long-term follow-up of Epstein-Barr virus viremia in pediatric recipients of renal transplants

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Survival Signals and Targets for Therapy in Breast Implant–Associated ALK⁻Anaplastic Large Cell Lymphoma