Epstein-Barr Virus Load In Patients Research Articles

Long term treatment with abatacept or tocilizumab does not increase Epstein-Barr virus load in patients with rheumatoid arthritis - A three years retrospective study.

BackgroundEpstein-Barr Virus (EBV) is a widely disseminated lymphotropic herpes virus implicated in benign and malignant disorders. In transplant patients, immunosuppressive drugs (cyclosporine) diminish control of EBV replication, potentially leading to lymphoproliferative disorders (LPD). Rheumatoid arthritis (RA) patients have impaired control of EBV infection and have EBV load ten times higher than controls. As post transplant patients, patients with RA have increased risk of developing lymphomas. Immunosuppressive drugs used to treat RA (conventional disease modifying drugs cDMARDs or biologics bDMARDs) could enhance the risk of developing LPD in RA patients. We have previously shown that long term treatment with Methotrexate and/or TNF alpha antagonists does not increase EBV load in RA. Our objective was to monitor the Epstein-Barr Virus load in RA patients treated with Abatacept (CTLA4 Ig), a T cell coactivation inhibitor, and Tocilizumab, an anti IL6 receptor antibody.MethodsEBV load in the peripheral blood mononuclear cells (PBMCs) of 55 patients under Abatacept (in 34% associated with Methotrexate) and 35 patients under Tocilizumab (in 37% associated with Methotrexate) was monitored for durations ranging from 6 months to 3 years by real time PCR. The influences of treatment duration and disease activity score 28 (DAS28) index on EBV load were analyzed.ResultsAbatacept did not significantly modify EBV load over time. Tocilizumab significantly diminished EBV load over time. No patient (of 90) developed EBV associated lymphoma.ConclusionLong term treatment with Abatacept or Tocilizumab does not increase EBV load in the PBMNCs of patients with RA.

TNFα antagonist therapy does not increase the Epstein-Barr virus burden in patients with rheumatoid arthritis or ankylosing spondylitis

ObjectiveThe risk of non-Hodgkin lymphoma is increased in rheumatoid arthritis (RA) but not in ankylosing spondylitis (AS). In RA, the degree of inflammation is closely associated with the lymphoma risk. Whether immunosuppressants such as methotrexate and TNFα antagonists affect the lymphoma risk in RA is unclear. The Epstein-Barr virus (EBV) may contribute to the pathogenesis of RA and may be involved in the development of lymphoma in patients taking methotrexate and/or TNFα antagonists, although these points remain debated. EBV load monitoring during immunosuppressive treatment may predict the occurrence of EBV-related lymphoma. Here, our objective was to prospectively measure the EBV load in patients receiving TNFα antagonists for RA or AS. MethodsWe prospectively studied patients with RA or AS before and after TNFα antagonist therapy initiation. The EBV load was measured in blood samples using the EBV R-gene Quantification Kit. Disease activity at the time of blood sampling was evaluated by determining the Disease Activity Score 28 (DAS28) in RA patients and the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) in AS patients. ResultsWe included 46 patients with RA (82.6% women; mean age, 52.7±11.3 years) and 27 with AS (men, 81.5%; mean age, 45.1±12.7 years). In the RA group, the EBV load was measured at baseline and 9.72±5.7 months later. The baseline EBV load was undetectable in 33 (70.2%) patients; mean EBV load in the 13 remaining patients was 9389copies/ml (3.47 log10±0.45). Baseline EBV load did not correlate with disease activity (DAS28). At the follow-up assay, the EBV load became positive in five patients and increased significantly in one patient (four patients on etanercept, one on adalimumab, and one on infliximab); it became negative in six patients (five on adalimumab and one on etanercept) and showed non-significant changes in six patients. Mean EBV load in patients positive at follow-up was 3.63±0.52 log10 copies/ml. Mean DAS28 was 4.78±1.1 at baseline and 2.94±1.24 at follow-up. At follow-up, a good EULAR response was noted in 33 (71.7%) patients and a moderate EULAR response in seven (15.2%) patients. In the AS group, the baseline EBV load measurement occurred after 12.9±10.6 months. Baseline EBV load was undetectable in 25 (92.6%) patients; mean load in the remaining two patients was 4.15±0.46 log10 copies/ml. At follow-up, the EBV load became positive in two patients (one on adalimumab and one on infliximab) and became negative in one patient (on adalimumab). Mean load in positive patients was 3.33±0.24 log10 copies/ml. Mean BASDAI was 55.1±16.2 at baseline and 17.88±18.62 at follow-up. A positive EBV load was significantly more common in the RA group than in the AS group (P=0.039). EBV load changes did not differ significantly between the RA and AS groups or across TNFα antagonists. No cases of lymphoma were recorded. ConclusionIntroducing TNFα antagonist therapy does not affect the EBV load in patients with RA or AS. EBV load monitoring is probably unnecessary in patients given TNFα antagonists for RA or AS.

Tonsillar CD38+ Activated B Cell Fraction Harbours the Highest EBV Load in Patients with Infectious Mononucleosis.

Epstein-Barr virus (EBV) persists within the B cell system and, in the blood of healthy virus carriers, is sequestered in the immunoglobulin (Ig)Dneg, CD27+ (“class-switched”) memory B cell compartment. This selective colonisation is apparent even in the blood of infectious mononucleosis (IM) patients undergoing primary EBV infection. Exactly how this is achieved is not understood, one theory being that the virus infects naïve B cells preferentially in vivo and drives them into memory using the physiologic process of germinal centre (GC) transit within lymphoid tissues. Here we have studied the distribution of EBV-infected B cells in tonsil cell preparations from acute IM patients and from chronic carriers. The following subsets of tonsillar B cells were isolated to high purity by FACS sorting and their EBV DNA loads were then determined by quantitative PCR: Naïve (CD38neg, IgD+, CD27neg); Class-switched memory (CD38neg, IgDneg, CD27+); Non-switched memory (CD38neg, IgD+, CD27+); Germinal centre (CD38+); and Plasma cell (CD38hi). In 15 carrier tonsils, the highest EBV load was consistently detected in class-switched memory, followed by significant amounts in non-switched memory. However, viral load in the CD38+ GC subset was usually very low, comparable to that in the naïve B cell fraction. This contrasted with the situation in 7 IM tonsils where overall viral loads were on average 100-fold higher than above. Here most viral DNA was consistently found in the CD38+ fraction, with lower loads in the class-switched memory and the smaller non-class switched memory subsets. We then investigated further the phenotype of the CD38+ population and found that, in IM tonsils, this not only included cells with the CD10 and CD77 markers of germinal centre origin but also other cells lacking these markers. Adopting a different FACS sorting strategy based on CD10, CD77 and IgD staining, we found that the virus was not present at significant levels in either the centrocyte (CD10+ CD77neg) or centroblast (CD77+) fractions. Thus EBV appeared to be concentrated in B cells expressing CD38 as a marker of activation rather than of germinal centre origin; this concurred with the results of immunohistochemical staining for the EBER RNAs showing an extrafollicular distribution of virus-infected cells in these IM tonsils. Our findings suggest that CD38 expression is a useful marker for B cells recently activated by EBV infection in vivo and show that, during primary EBV infection, the tonsillar CD38+ B cells lacking germinal centre markers harbour the highest viral load. If EBV does exploit the germinal centre pathway as an entry route into B cell memory, where primary infection is manifest as IM this pathway must be taken by a very small fraction of infected B cells and/or must have been completed before disease symptoms appear.

A longitudinal and prospective study of Epstein-Barr virus load in AIDS-related non-Hodgkin lymphoma

Background Epstein-Barr virus (EBV) may be causally associated with non-Hodgkin Lymphoma (NHL) in HIV-infected patients. Objectives To compare EBV load in whole blood in AIDS-NHL patients, HIV non-AIDS patients and non-HIV-infected persons, and to prospectively measure EBV load in whole blood in AIDS-NHL patients. Study design Longitudinal and prospective study. Results We observed no statistical difference in EBV load between AIDS-NHL (3.69 log 10 copies/mL [interquartile range (IQR): 2.89–4.27]) and HIV non-AIDS patients (3.08 log 10 copies/mL [IQR: 1.29–3.57]) but AIDS-NHL patients had significantly higher EBV loads than HIV-negative controls (1.19 log 10 copies/mL [IQR: 0.00–3.29]). We noticed an inverse correlation between CD4+ lymphocytes count and EBV load in patients with AIDS-NHL ( r 2 = 0.41, P = 0.01). In the longitudinal study, the mean EBV load three months after NHL diagnosis decreased significantly (mean difference = −1.69 log 10 copies/mL [95% confidence interval: −0.32; −3.04]; P = 0.03) under chemotherapy but was still elevated in patients with relapses or no response to chemotherapy. Conclusion Although EBV load seems a suboptimal marker for the diagnosis of AIDS-NHL, we observed a significant decrease of EBV load in patients treated with chemotherapy and a strong association between NHL outcome and EBV load in whole blood.

Normalized quantification by real-time PCR of Epstein-Barr virus load in patients at risk for posttransplant lymphoproliferative disorders.

The load of Epstein-Barr virus (EBV) in peripheral blood mononuclear cells of transplant recipients represents a predictive parameter for posttransplant lymphoproliferative disorders (PTLD). The aim of our work was to develop a rapid and reliable PCR protocol for the quantification of cell-associated EBV DNA in transplant recipients. In contrast to previous studies, a protocol that facilitated quantification independent of photometric nucleic acid analysis was established. We took advantage of the real-time PCR technology which allows for single-tube coamplification of EBV and genomic C-reactive protein (CRP) DNA. EBV copy numbers were normalized by division by the amount of CRP DNA, with the quotient representing the actual amount of amplifiable genomic DNA per reaction. Coamplification of CRP DNA did not result in a diminished detection limit for EBV. By using the protocol without normalization, EBV copy numbers in 4 out of 10 PTLD patients were within the normal range determined with data for 114 transplant recipients that served as controls. After normalization, however, all of the PTLD patients had a higher viral load than the control population, indicating an increased sensitivity of the assay. Moreover, EBV copy numbers obtained for one patient by conventional quantification and suggestive of relapsing PTLD were within normal range after normalization. We conclude that normalization of PCR signals to coamplified genomic DNA allows a more accurate quantification of cell-bound EBV.