Effect of methotrexate and anti-TNF on Epstein-Barr virus T-cell response and viral load in patients with rheumatoid arthritis or spondylarthropathies
- Corinne Miceli-Richard†1, 2,
- Nicolas Gestermann†2,
- Corinne Amiel3,
- Jérémie Sellam1, 2,
- Marc Ittah2,
- Stephan Pavy1,
- Alejandra Urrutia2,
- Isabelle Girauld2,
- Guislaine Carcelain4,
- Alain Venet2 and
- Xavier Mariette1, 2Email author
© Miceli-Richard et al.; licensee BioMed Central Ltd. 2009
Received: 24 December 2008
Accepted: 26 May 2009
Published: 26 May 2009
There is a suspicion of increased risk of Epstein-Barr virus (EBV)-associated lymphoproliferations in patients with inflammatory arthritides receiving immunosuppressive drugs. We investigated the EBV load and EBV-specific T-cell response in patients treated with methotrexate (MTX) or anti-TNF therapy.
Data for patients with rheumatoid arthritis (RA) (n = 58) or spondylarthropathy (SpA) (n = 28) were analyzed at baseline in comparison with controls (n = 22) and after 3 months of MTX or anti-TNF therapy for EBV load and EBV-specific IFNγ-producing T cells in response to EBV latent-cycle and lytic-cycle peptides.
The EBV load and the number of IFNγ-producing T-cells after peptide stimulation were not significantly different between groups at baseline (P = 0.61 and P = 0.89, respectively). The EBV load was not significantly modified by treatment, for RA with MTX (P = 0.74) or anti-TNF therapy (P = 0.94) or for SpA with anti-TNF therapy (P = 1.00). The number of EBV-specific T cells was not significantly modified by treatment, for RA with MTX (P = 0.58) or anti-TNF drugs (P = 0.19) or for SpA with anti-TNF therapy (P = 0.39). For all patients, the EBV load and EBV-specific T cells were significantly correlated (P = 0.017; R = 0.21). For most patients, short-term exposure (3 months) to MTX or anti-TNF did not alter the EBV load or EBV-specific T-cell response but two patients had discordant evolution.
These data are reassuring and suggest there is no short-term defect in EBV-immune surveillance in patients receiving MTX or anti-TNF drugs. However, in these patients, long term follow-up of EBV-specific T-cell response is necessary and the role of non-EBV-related mechanisms of lymphomagenesis is not excluded.
Rheumatoid arthritis (RA) is associated with a twofold increase of non-Hodgkin's lymphoma  and a threefold increase of Hodgkin's lymphoma . The effect of immunosuppressive drugs on the risk of lymphoma is debated. Most recent studies did not find an overall increased risk of non-Hodgkin's lymphoma in RA patients treated with methotrexate (MTX). Several reports, however, showed that MTX can rarely induce Epstein-Barr virus (EBV)-associated lymphoproliferation regressive after withdrawal of the drug [3, 4].
Recent concerns about possible treatment effects and lymphoma have focused on anti-TNF drugs because of their profound immunoregulatory effect. A recent meta-analysis of randomized controlled trials of infliximab and adalimumab identified 10 cases of lymphoma (four cases in the randomized phase of the trials and six cases in the extension phase) in the treated groups (3,493 patient-years) and none in the placebo groups (1,512 patient-years) . Inflammatory activity of the underlying disease is the main risk factor of lymphoma in RA , however, and anti-TNF therapy is used for patients with the most active disease. Results for three large cohorts of RA patients did not reveal any increased risk of lymphoma in RA patients receiving anti-TNF drugs versus RA patients receiving classical disease-modifying anti-rheumatic drugs (DMARDs). In most of these cohorts, however, increased risk of lymphoma persisted as compared with that in the general population [7–9].
Cases of EBV-associated lymphoproliferation that regressed after withdrawal of MTX have been described [3, 4]. Case reports of lymphoma associated or not with EBV, treated with anti-TNF drugs and regressing after withdrawal of therapy have also been reported [10, 11]. These cases may mimic post-transplant lymphoproliferative disorder, a severe complication of EBV reactivation linked to impaired EBV control by CD8 T cells and arising in allograft recipients receiving immunosuppressive drugs .
Taken together, such data provide reliable arguments to investigate a potential EBV reactivation during MTX and/or TNFα antagonist therapy as a possible first step of lymphoma induction. During primary EBV infection, specific cytotoxic CD8+ T cells expand and recognize epitopes from lytic-cycle antigens and, to a lesser extent, from latent-cycle antigens. A small population of EBV-specific memory CD8+ T cells further persists  and plays a crucial role in the control of persistent EBV infection . An impaired EBV-specific T-cell response could constitute one of the first steps of lymphoma induction with immunosuppressive drug therapy.
The present study aimed to determine the EBV viral load and the specific effector CD8+ T-cell response against EBV antigens in patients with RA and spondylarthropathy (SpA) receiving MTX or anti-TNF drugs, to shed some light on a possible impaired EBV-specific T-cell response as the triggering mechanism of lymphomagenesis in this population.
Materials and methods
All studied subjects were seropositive for EBV. The present study consisted of two parts. In the cross-sectional first part of the study we investigated EBV-specific IFNγ-producing T cells at baseline (week 0) in 87 patients: 32 MTX naïve RA patients (mean age 60 ± 16 years, mean duration of disease 4.5 ± 6.6 years), 27 patients with RA receiving MTX who were not responders to the drug (mean age 53 ± 11 years, mean duration of disease 9.5 ± 10.5 years) and 28 patients with SpA (14 not receiving DMARDs and 14 receiving MTX; mean age 36 ± 11 years, mean duration of disease 9.6 ± 9.7 years). Patients with RA fulfilled the 1987 American College of Rheumatology criteria  and those with SpA fulfilled the European Spondylarthropathy Study Group criteria . All RA patients were rheumatoid factor positive and/or anti-cyclic citrullinated peptide positive. The Disease Activity Score for 28 joints was 4.8 ± 1.2 in naïve RA patients and was 5.6 ± 1.2 in RA patients who were nonresponders to MTX. The Bath Ankylosing Spondylitis Disease Activity Index score  was 55 ± 22 in SpA patients. The control group comprised 22 patients with mechanic radiculopathic conditions (mean age 47 ± 15 years).
From the 87 patients included in the cross-sectional part of the study, 62 underwent the second longitudinal part of the study for EBV-specific IFNγ-producing T cells after 3 months (week 12) of MTX or anti-TNF treatment. Forty patients (21 SpA and 19 RA) received anti-TNF drugs. All RA patients and 10/21 SpA patients had anti-TNF + MTX. Twenty-two MTX naive RA patients received MTX. EBV viral load data were also available for 67 patients and 15 control individuals at week 0, and for 52 patients at week 12.
The present study was performed with approval of the local ethics committee (CPP Ile de France 7), and informed consent was obtained from all study participants.
Isolation of peripheral blood mononuclear cells
Peripheral blood mononuclear cells (PBMCs) were isolated by density gradient centrifugation using Ficoll-Hypaque 1.107 (Biochrom, Berlin, Germany). The PBMCs were then frozen in FCS containing 10% dimethyl sulfoxide (Sigma, Saint-Quentin Fallavier, France) and stored in liquid nitrogen until use.
Epstein-Barr virus peptides
Human leukocyte antigen class I-restricted cytotoxic T-lymphocyte Epstein-Barr virus epitopes
Human leukocyte antigen
596 to 604
284 to 293
329 to 337
426 to 434
280 to 288
453 to 461
603 to 611
399 to 408
416 to 424
340 to 350
131 to 139
246 to 253
419 to 427
217 to 225
442 to 451
491 to 499
176 to 184
502 to 510
379 to 387
881 to 889
158 to 166
325 to 333
190 to 197
244 to 254
258 to 266
236 to 244
149 to 157
249 to 258
343 to 351
458 to 466
488 to 496
407 to 417
271 to 278
567 to 666
281 to 290
335 to 343
163 to 171
200 to 208
406 to 414
831 to 839
213 to 222
The ELISPOT-IFNγ assay was used to determine the frequency of T cells that produced IFNγ in response to a brief exposure to EBV antigens, as previously published . Briefly, nitrocellulose ELISPOT plates (Millipore, Guyancourt, France) were coated with anti-IFNγ antibody (1 μg/ml, 100 μl/well in PBS; 1-D1K; Mabtech, Sophia Antipolis, France). PBMCs were added in duplicate wells at 105 cells per well with 2 μg/ml peptide. The second biotinylated anti-IFNγ monoclonal antibody was then added (7-B6-biotin; Mabtech) and IFNγ secreting cells were revealed with an enzymatic reaction with streptavidin-conjugated alkaline phosphatase (Sigma-Aldrich, Saint-Quentin Fallavier, France).
The number of specific T-cell responders per 106 PBMCs was calculated after subtraction of the background, which corresponded to the mean value of IFNγ spots associated with nonstimulated PBMCs (PBMCs in the presence of medium alone). Results were expressed as spot-forming cells (SFCs) per 106 PBMCs and were calculated for each pool of peptides as follows:
SFCs/106 PBMCs = 10 x (mean SFCs/105 cells from two antigen-stimulated wells - mean SFC/105 cells from four unstimulated wells).
Results were presented as the individual response to the set of latent-cycle peptides (9-mer peptides), to the set of lytic-cycle peptides (BMLF 9-mer peptide added with the 15-mer lytic-cycle peptides) or to both sets.
Wells were counted as positive if they contained at least 50 SFCs/106 PBMCs and exhibited at least twofold the mean value of the background (per million PBMC). The median number of IFNγ-producing PBMCs in the presence of medium alone (background) was zero spots/well (range 0 to 4).
Epstein-Barr virus load in peripheral blood mononuclear cells
The level of EBV DNA copies in PBMCs was measured by Taqman real-time quantitative PCR as previously described . For each quantification, 500,000 to 106 PBMCs were thawed and DNA extractions were further performed. The PCR primers were selected to amplify a 121 bp product in the thymidine kinase gene. A pcDNA 3.1 vector (Invitrogen, Groningen, the Netherlands) containing one copy of the EBV target region was used as standard for EBV quantification. The level of albumin DNA copies in PBMC samples estimated by real-time PCR was used as the endogenous reference to normalize the variations in PBMC number or DNA extraction. All standard dilutions, control samples and PBMC samples were run in parallel and in duplicate for EBV and albumin DNA quantifications. The normalized value of the cell-associated EBV DNA load corresponding to the ratio EBV average copy number/albumin average copy number × 2.106 was finally expressed as the number of EBV DNA copies per 106 PBMC.
Results are given as the percentage of patients with positive EBV T-cell response, as well as the mean response ± standard deviation. Statistical analyses involved use of StatView 5.0 (Abacus Concepts, Berkeley, CA, USA). Nonparametric tests were used. Comparisons between groups involved the Kruskal-Wallis test. Cross-sectional comparison of EBV T spots or the EBV copy number distribution involved the Mann-Whitney rank-sum test. Longitudinal comparison of EBV T spots or the EBV copy number between week 0 and week 12 involved the Wilcoxon test. Correlation studies involved Spearman's correlation. P < 0.05 was considered statistically significant.
Epstein-Barr virus load in peripheral blood mononuclear cells
We found no significant correlation between the EBV viral load and disease activity (Disease Activity Score for 28 joints for RA patients, P = 0.54; Bath Ankylosing Spondylitis Disease Activity Index for SpA patients, P = 0.84) or disease duration (P = 0.29).
Epstein-Barr virus-specific IFNγ-producing T cells
We found no significant correlation between the number of EBV-specific IFNγ-producing T cells and disease activity (Disease Activity Score for 28 joints for RA patients (n = 64), P = 0.32; Bath Ankylosing Spondylitis Disease Activity Index for SpA patients (n = 21), P = 0.47) or disease duration (n = 88, P = 0.40).
Epstein-Barr virus load in peripheral blood mononuclear cells
Epstein-Barr virus-specific IFNγ-producing T cells
In response to the full set of peptides, the number of IFNγ-producing cells was not significantly modified by immunosuppressive treatment (SpA patients receiving anti-TNF drugs (n = 21), P = 0.39; RA patients receiving anti-TNF drugs (n = 19), P = 0.19; RA patients receiving MTX (n = 22), P = 0.58) (Figure 3b), nor was the number of EBV-specific IFNγ-producing T cells modified when considering each set of peptides (latent-cycle peptides or lytic-cycle peptides) (data not shown). Among patients treated with TNF blockers, there was no difference according to the class of molecule used: monoclonal antibody (infliximab and adalimumab) (n = 16, P = 0.74) or soluble receptor (etanercept) (n = 24, P = 0.92).
Correlation between Epstein-Barr virus load and T spots
Unadapted Epstein-Barr virus-specific T-cell IFNγ production under treatment
The present large cross-sectional and longitudinal study showed no abnormality in EBV viral load or EBV-specific T-cell response in patients with RA or SpA at baseline or after treatment with MTX or anti-TNF drugs.
In contrast to other studies [23, 24], we did not find increased EBV viral load in PBMCs of patients with RA or SpA. A casual high EBV DNA prevalence in our control group (80%) could account for the differing results, and/or the highly sensitive PCR used in our study might explain such differences. Our cross-sectional study revealed no significant differences between patients and control individuals in the proportion of subjects with positive EBV-specific IFNγ-producing T cells in PBMCs, the mean number of SFCs or the SFC distribution. Two studies have assessed the EBV-specific T-cell response in PBMCs of RA patients [25, 26]. The first study found no difference between 49 RA patients and 26 control individuals in the frequency of T cells directed against two immunodominant EBV peptides, but did observe a reduced ability to produce INFγ in RA patients; the effect of immunosuppressive treatment was not assessed . In the second study, EBV-specific effector CD8 T cells were higher in number in RA patients (n = 25) than in control individuals (n = 20), but this study concerned a low number of patients and was only cross-sectional . Actually, this increase in IFNγ secreted T cells was related to increased viral load, which we did not observe in our study.
Our longitudinal results did not reveal any influence of immunosuppressive treatment on the EBV viral load. These results are in accordance with several studies on Crohn disease or RA patients assessing the evolution of EBV viral load under immunosuppressive treatment [27–29]. To the best of our knowledge, no published study has specifically evaluated the longitudinal effect of MTX and anti-TNF drug on EBV-specific T-cell effector functions in patients with RA or SpA. At 3-month follow-up, neither MTX treatment in RA patients nor anti-TNF therapy in RA and/or SpA patients modified these effector functions, regardless of the EBV peptide used for pulsing – latent-cycle peptides or lytic-cycle peptides or the full set of peptides. The lack of increase in the EBV viral load during the same period in all groups of patients agrees with the preserved specific T-cell effector function, which was confirmed by a global correlation between EBV viral load and EBV-specific T-cell response.
Interestingly, in five patients treated with anti-TNF, an inadequate in vitro EBV-specific IFNγ production was observed after specific pulse with EBV peptides despite an in vivo high viral load. Among those patients, two different profiles were observed. The first profile corresponded to patients without any IFNγ production in spite of high EBV viral loads (>1,000/106 PBMCs), both at week 0 and week 12. In such cases, the lack of adequate HLA for presenting one of the EBV peptides probably accounted for the absence of IFNγ production after specific pulse. The second profile corresponded to two patients having EBV-specific T-cell IFNγ production at baseline but a discordant evolution between an IFNγ secreted T-cell decrease and an EBV viral load increase after 12 weeks of treatment. In these two patients, immunosuppressive therapy might have impaired EBV-specific T-cell effector functions leading to the lack of control of the EBV viral load. These two patients having been treated with the association of anti-TNF antibody and MTX makes it impossible to differentiate a possible effect of one drug individually. Nevertheless, we never saw profound discrepancies, such as those observed in a pediatric sample in whom post-transplant lymphoproliferative disorder developed after liver transplantation . Since we analyzed data only 12 weeks after the introduction of immunosuppressive treatment, however, we cannot exclude that MTX or anti-TNF therapy could induce impaired EBV control after longer-term treatment. This relative short time duration of immunosuppressive treatment exposure might be considered as a limitation of our study. Nevertheless, post-transplant lymphoproliferative diseases occurring in children, for example, have been reported to occur after a short-term exposure to immunosuppressive treatments (median delay of 12 weeks, range 6 to 56 weeks) . Moreover, with the same methodology (ELISPOT assay), we detected a significant decrease of the specific anti-tuberculosis T-cell response in patients after 12 weeks of anti-TNF therapy . Lastly, in the present study, patients treated with MTX were treated for several years on average, and their results were no different from the MTX naïve patients at baseline.
In patients with RA or SpA, short-term (3-month) exposure to MTX or anti-TNF therapy does not alter the EBV viral load or the EBV-specific T-cell response. These findings are rather reassuring in light of a suggested increased risk of EBV-associated lymphoma in patients receiving immunosuppressive therapy. Long-term follow-up of the EBV-specific T-cell response, however, is necessary. Moreover, control of EBV is only one mechanism of control of lymphomagenesis and the different epidemiologic studies currently available do not eliminate the possibility of increased risk of non-EBV-associated lymphoma in patients receiving immunosuppressive therapy.
disease-modifying anti-rheumatic drug
fetal calf serum
human leukocyte antigen
peripheral blood mononuclear cell
polymerase chain reaction
tumor necrosis factor.
The set of EBV peptides was kindly provided by Prof. D Olive (INSERM Action-Thématique-Concertée). The authors thank Dr Martine Sinet and Prof. Martine Raphael for helpful discussions. The present work was supported by Assistance Publique Hôpitaux de Paris, Département de la Recherche Clinique, Paris, France (Contrat d'Initiation à la Recherche Clinique) and by Société Française de Rhumatologie.
- Smedby KE, Hjalgrim H, Askling J, Chang ET, Gregersen H, Porwit-MacDonald A, Sundstrom C, Akerman M, Melbye M, Glimelius B, Adami HO: Autoimmune and chronic inflammatory disorders and risk of non-Hodgkin lymphoma by subtype. J Natl Cancer Inst. 2006, 98: 51-60.View ArticlePubMedGoogle Scholar
- Landgren O, Engels EA, Pfeiffer RM, Gridley G, Mellemkjaer L, Olsen JH, Kerstann KF, Wheeler W, Hemminki K, Linet MS, Goldin LR: Autoimmunity and susceptibility to Hodgkin lymphoma: a population-based case-control study in Scandinavia. J Natl Cancer Inst. 2006, 98: 1321-1330.View ArticlePubMedGoogle Scholar
- Kamel OW, Rijn van de M, Weiss LM, Del Zoppo GJ, Hench PK, Robbins BA, Montgomery PG, Warnke RA, Dorfman RF: Brief report: reversible lymphomas associated with Epstein-Barr virus occurring during methotrexate therapy for rheumatoid arthritis and dermatomyositis. N Engl J Med. 1993, 328: 1317-1321. 10.1056/NEJM199305063281806.View ArticlePubMedGoogle Scholar
- Salloum E, Cooper DL, Howe G, Lacy J, Tallini G, Crouch J, Schultz M, Murren J: Spontaneous regression of lymphoproliferative disorders in patients treated with methotrexate for rheumatoid arthritis and other rheumatic diseases. J Clin Oncol. 1996, 14: 1943-1949.PubMedGoogle Scholar
- Bongartz T, Sutton AJ, Sweeting MJ, Buchan I, Matteson EL, Montori V: Anti-TNF antibody therapy in rheumatoid arthritis and the risk of serious infections and malignancies: systematic review and meta-analysis of rare harmful effects in randomized controlled trials. JAMA. 2006, 295: 2275-2285. 10.1001/jama.295.19.2275.View ArticlePubMedGoogle Scholar
- Baecklund E, Iliadou A, Askling J, Ekbom A, Backlin C, Granath F, Catrina AI, Rosenquist R, Feltelius N, Sundstrom C, Klareskog L: Association of chronic inflammation, not its treatment, with increased lymphoma risk in rheumatoid arthritis. Arthritis Rheum. 2006, 54: 692-701. 10.1002/art.21675.View ArticlePubMedGoogle Scholar
- Wolfe F, Michaud K: Lymphoma in rheumatoid arthritis: the effect of methotrexate and anti-tumor necrosis factor therapy in 18,572 patients. Arthritis Rheum. 2004, 50: 1740-1751. 10.1002/art.20311.View ArticlePubMedGoogle Scholar
- Geborek P, Bladstrom A, Turesson C, Gulfe A, Petersson IF, Saxne T, Olsson H, Jacobsson LT: Tumour necrosis factor blockers do not increase overall tumour risk in patients with rheumatoid arthritis, but may be associated with an increased risk of lymphomas. Ann Rheum Dis. 2005, 64: 699-703. 10.1136/ard.2004.030528.PubMed CentralView ArticlePubMedGoogle Scholar
- Schiff MH, Burmester GR, Kent JD, Pangan AL, Kupper H, Fitzpatrick SB, Donovan C: Safety analyses of adalimumab (HUMIRA) in global clinical trials and US postmarketing surveillance of patients with rheumatoid arthritis. Ann Rheum Dis. 2006, 65: 889-894. 10.1136/ard.2005.043166.PubMed CentralView ArticlePubMedGoogle Scholar
- Park SH, Kim CG, Kim JY, Choe JY: Spontaneous regression of EBV-associated diffuse lymphoproliferative disease in a patient with rheumatoid arthritis after discontinuation of etanercept treatment. Rheumatol Int. 2008, 28: 475-477. 10.1007/s00296-007-0467-6.View ArticlePubMedGoogle Scholar
- Thonhofer R, Gaugg M, Kriessmayr M, Neumann HJ, Erlacher L: Spontaneous remission of marginal zone B cell lymphoma in a patient with seropositive rheumatoid arthritis after discontinuation of infliximab-methotrexate treatment. Ann Rheum Dis. 2005, 64: 1098-1099. 10.1136/ard.2004.026252.PubMed CentralView ArticlePubMedGoogle Scholar
- Boubenider S, Hiesse C, Goupy C, Kriaa F, Marchand S, Charpentier B: Incidence and consequences of post-transplantation lymphoproliferative disorders. J Nephrol. 1997, 10: 136-145.PubMedGoogle Scholar
- Hislop AD, Annels NE, Gudgeon NH, Leese AM, Rickinson AB: Epitope-specific evolution of human CD8(+) T cell responses from primary to persistent phases of Epstein-Barr virus infection. J Exp Med. 2002, 195: 893-905. 10.1084/jem.20011692.PubMed CentralView ArticlePubMedGoogle Scholar
- Dunne PJ, Faint JM, Gudgeon NH, Fletcher JM, Plunkett FJ, Soares MV, Hislop AD, Annels NE, Rickinson AB, Salmon M, Akbar AN: Epstein-Barr virus-specific CD8(+) T cells that re-express CD45RA are apoptosis-resistant memory cells that retain replicative potential. Blood. 2002, 100: 933-940. 10.1182/blood-2002-01-0160.View ArticlePubMedGoogle Scholar
- Arnett FC, Edworthy SM, Bloch DA, McShane DJ, Fries JF, Cooper NS, Healey LA, Kaplan SR, Liang MH, Luthra HS, Medsger TA, Mitchell DM, Neustadt DH, Pinals RS, Schaller JG, Sharp JT, Wilder RL, Hunder GG: The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. Arthritis Rheum. 1988, 31: 315-324. 10.1002/art.1780310302.View ArticlePubMedGoogle Scholar
- Dougados M, Linden van der S, Juhlin R, Huitfeldt B, Amor B, Calin A, Cats A, Dijkmans B, Olivieri I, Pasero G, Veys E, Zeidler H: The European Spondylarthropathy Study Group preliminary criteria for the classification of spondylarthropathy. Arthritis Rheum. 1991, 34: 1218-1227. 10.1002/art.1780341003.View ArticlePubMedGoogle Scholar
- Garrett S, Jenkinson T, Kennedy LG, Whitelock H, Gaisford P, Calin A: A new approach to defining disease status in ankylosing spondylitis: the Bath Ankylosing Spondylitis Disease Activity Index. J Rheumatol. 1994, 21: 2286-2291.PubMedGoogle Scholar
- Samri A, Durier C, Urrutia A, Sanchez I, Gahery-Segard H, Imbart S, Sinet M, Tartour E, Aboulker JP, Autran B, Venet A: Evaluation of the interlaboratory concordance in quantification of human immunodeficiency virus-specific T cells with a gamma interferon enzyme-linked immunospot assay. Clin Vaccine Immunol. 2006, 13: 684-697. 10.1128/CVI.00387-05.PubMed CentralView ArticlePubMedGoogle Scholar
- Yang J, Lemas VM, Flinn IW, Krone C, Ambinder RF: Application of the ELISPOT assay to the characterization of CD8(+) responses to Epstein-Barr virus antigens. Blood. 2000, 95: 241-248.PubMedGoogle Scholar
- Steven NM, Annels NE, Kumar A, Leese AM, Kurilla MG, Rickinson AB: Immediate early and early lytic cycle proteins are frequent targets of the Epstein-Barr virus-induced cytotoxic T cell response. J Exp Med. 1997, 185: 1605-1617. 10.1084/jem.185.9.1605.PubMed CentralView ArticlePubMedGoogle Scholar
- Doisne JM, Urrutia A, Lacabaratz-Porret C, Goujard C, Meyer L, Chaix ML, Sinet M, Venet A: CD8+ T cells specific for EBV, cytomegalovirus, and influenza virus are activated during primary HIV infection. J Immunol. 2004, 173: 2410-2418.View ArticlePubMedGoogle Scholar
- Besson C, Amiel C, Le-Pendeven C, Brice P, Ferme C, Carde P, Hermine O, Raphael M, Abel L, Nicolas JC: Positive correlation between Epstein-Barr virus viral load and anti-viral capsid immunoglobulin G titers determined for Hodgkin's lymphoma patients and their relatives. J Clin Microbiol. 2006, 44: 47-50. 10.1128/JCM.44.1.47-50.2006.PubMed CentralView ArticlePubMedGoogle Scholar
- Balandraud N, Meynard JB, Auger I, Sovran H, Mugnier B, Reviron D, Roudier J, Roudier C: Epstein-Barr virus load in the peripheral blood of patients with rheumatoid arthritis: accurate quantification using real-time polymerase chain reaction. Arthritis Rheum. 2003, 48: 1223-1228. 10.1002/art.10933.View ArticlePubMedGoogle Scholar
- Alvarez-Lafuente R, Fernandez-Gutierrez B, de Miguel S, Jover JA, Rollin R, Loza E, Clemente D, Lamas JR: Potential relationship between herpes viruses and rheumatoid arthritis: analysis with quantitative real time polymerase chain reaction. Ann Rheum Dis. 2005, 64: 1357-1359. 10.1136/ard.2004.033514.PubMed CentralView ArticlePubMedGoogle Scholar
- Klatt T, Ouyang Q, Flad T, Koetter I, Buhring HJ, Kalbacher H, Pawelec G, Muller CA: Expansion of peripheral CD8+ CD28-T cells in response to Epstein-Barr virus in patients with rheumatoid arthritis. J Rheumatol. 2005, 32: 239-251.PubMedGoogle Scholar
- Lunemann JD, Frey O, Eidner T, Baier M, Roberts S, Sashihara J, Volkmer R, Cohen JI, Hein G, Kamradt T, Munz C: Increased frequency of EBV-specific effector memory CD8+ T cells correlates with higher viral load in rheumatoid arthritis. J Immunol. 2008, 181: 991-1000.PubMed CentralView ArticlePubMedGoogle Scholar
- Reijasse D, Le Pendeven C, Cosnes J, Dehee A, Gendre JP, Nicolas JC, Beaugerie L: Epstein-Barr virus viral load in Crohn's disease: effect of immunosuppressive therapy. Inflamm Bowel Dis. 2004, 10: 85-90. 10.1097/00054725-200403000-00004.View ArticlePubMedGoogle Scholar
- Torre-Cisneros J, Del Castillo M, Caston JJ, Castro MC, Perez V, Collantes E: Infliximab does not activate replication of lymphotropic herpesviruses in patients with refractory rheumatoid arthritis. Rheumatology (Oxford). 2005, 44: 1132-1135. 10.1093/rheumatology/keh696.View ArticleGoogle Scholar
- Balandraud N, Guis S, Meynard JB, Auger I, Roudier J, Roudier C: Long-term treatment with methotrexate or tumor necrosis factor alpha inhibitors does not increase Epstein-Barr virus load in patients with rheumatoid arthritis. Arthritis Rheum. 2007, 57: 762-767. 10.1002/art.22783.View ArticlePubMedGoogle Scholar
- Smets F, Latinne D, Bazin H, Reding R, Otte JB, Buts JP, Sokal EM: Ratio between Epstein-Barr viral load and anti-Epstein-Barr virus specific T-cell response as a predictive marker of posttransplant lymphoproliferative disease. Transplantation. 2002, 73: 1603-1610. 10.1097/00007890-200205270-00014.View ArticlePubMedGoogle Scholar
- Hamdi H, Mariette X, Godot V, Weldingh K, Hamid AM, Prejean MV, Baron G, Lemann M, Puechal X, Breban M, Berenbaum F, Delchier JC, Flipo RM, Dautzenberg B, Salmon D, Humbert M, Emilie D: Inhibition of anti-tuberculosis T-lymphocyte function with tumour necrosis factor antagonists. Arthritis Res Ther. 2006, 8: R114-10.1186/ar1994.PubMed CentralView ArticlePubMedGoogle Scholar
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