Rheumatoid Arthritis: Miossec P

Miossec P. · No affiliation provided · Joint Bone Spine. · Pubmed #15050191 No free full text.

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Miossec P. · Department of Immunology and Rheumatology, Hospital Edouard Herriot, University of Lyon, 69437 Lyon, France. · Arthritis Res Ther. · Pubmed #19007422 links to  free full text

Abstract: This review focuses on the contributions made by interactions between dendritic cells (DCs) and T cells, and by local production of cytokines and chemokines to the pathogenesis of rheumatoid arthritis (RA) synovitis. DCs are efficient professional antigen-presenting cells, which are critical for the development of innate and adaptative immune responses through interactions with T cells. Cytokines from DCs play a key role in the switch inside effector T-cell pathways. Chemokines are important mediators of the immune response because they regulate leucocyte recruitment to tissue, and they play a key role in inflammatory diseases by acting on T-cell and DC migration. Furthermore, the recently discovered T-helper-17 proinflammatory cytokines, present in syno-vium samples, are associated with the migration, differentiation and maturation of inflammatory cells, and they facilitate a network of interactions between all components of the immune response. An understanding of such interactions is essential because it is the key to therapeutic application.

Lafeber FP, Miossec P, Valentino LA. · Department of Rheumatology and Clinical Immunology, University Medical Center Utrecht, Utrecht, The Netherlands. · Haemophilia. · Pubmed #18494686 No free full text.

Abstract: Haemophilic arthropathy, which shares some clinical and biological injury characteristics with rheumatoid arthritis, is characterized by two main features: chronic proliferative synovitis and cartilage destruction. It is the consequence of repeated extravasation of blood into joint cavities, but its exact pathogenesis, particularly with regard to early changes in the joint, is still incompletely understood. This review presents recent findings obtained in experiments performed in vitro and using animal models, which have improved our knowledge of the pathogenesis of haemophilic arthropathy. These experimental studies show that haemophilic arthropathy is a multifactorial event in which the deposit of iron in the joints appears to exert a central role. First, iron may promote the apoptosis of chondrocytes by catalysing the formation of oxygen metabolites; this may explain the fact that intra-articular blood exerts a directly harmful effect on cartilage before, and independent of synovial changes. Secondly, iron may also act on the synovial membrane by favouring its proliferation through the induction of proto-oncogenes involved in cellular proliferation and stimulation of inflammatory cytokines as well as abrogation of apoptosis. These two processes, one degenerative and cartilage-mediated, the other inflammatory and synovium-mediated could occur in parallel or sequentially. Overall, it may be expected that these experimental results will yield new therapeutic strategies capable of effectively preventing the occurrence of this still serious and common complication in patients with severe haemophilia.

Miossec P. · Department of Immunology and Rheumatology, Hôpital Edouard Herriot, Lyon, France. · Arthritis Rheum. · Pubmed #17599728 links to  free full text

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Toh ML, Miossec P. · Department of Immunology and Rheumatology, Hôpital Edouard Herriot, Lyon, France. · Curr Opin Rheumatol. · Pubmed #17414957 No free full text.

Abstract: PURPOSE OF REVIEW: To update the knowledge on the contribution of T cells in rheumatoid arthritis, a selection of publications between the end of 2005 and 2006 were reviewed. RECENT FINDINGS: Th17 cells driven by TGF-beta, IL-1, IL-6 and IL-23 challenge previous concepts of 'Th1'-induced rheumatoid arthritis. Other advancements in IL-17 studies include novel concepts on the IL-17 receptor and additional information on the mechanism of IL-17-induced effects. Regulatory T cells fail to control disease due to defective function secondary to the synovial inflammatory milieu. The predominance of pathogenic effector T cells in the presence of impaired T-cell regulatory mechanisms may therefore contribute to rheumatoid arthritis chronicity. Cellular therapies attempt to restore the balance that includes production of immunoregulatory cytokines such as IL-4 or IL-10. Better T-cell-targeted therapies controlling costimulation are in place with purported increased efficacy and durability, including anti-tumour necrosis factor nonresponders. Additional direct and indirect T-cell approaches include antagonism of T-cell-derived cytokines, T-cell activation or B-cell ablation. SUMMARY: A renewed interest in T cells comes from the discovery of Th17 in rheumatoid arthritis and from novel findings on the role of T cells in rheumatoid arthritis induction, chronicity and relapse.

Miossec P. · Department of Immunology and Rheumatology, Hôpital Edouard Herriot, 69437 Lyon Cedex 03, France. · Curr Opin Rheumatol. · Pubmed #15103248 No free full text.

Abstract: PURPOSE OF REVIEW: To update the knowledge accumulated on the contribution of cytokines to rheumatoid arthritis and related animal models. Publications from the end of 2002 and 2003 period were analyzed for a selection. RECENT FINDINGS: A better understanding of the clinical results with tumor necrosis factor-alpha inhibitors has come from studies in treated patients. The expected effect of infliximab on the apoptosis of cells expressing tumor necrosis factor-alpha was not observed in synovium biopsy specimens. The mode of action of tumor necrosis factor-alpha on bone destruction has been clarified in gene-defective mice. Tumor necrosis factor-alpha acts through osteoclasts--an effect that is inhibited with osteoprotegerin. New interleukin-1 inhibitors with a potential for increased efficacy, such as interleukin-1trap, have been manufactured and are now being tested in rheumatoid arthritis. The list of cytokines of interest for therapeutic intervention has been growing rapidly. The results with animal models have provided clues to control arthritis with natural interleukin-18 inhibitors, such as interleukin-18 BP. Additional results have been accumulated that indicate the contribution of T cell subsets in inflammation and destruction through the production of interleukin-17. Synergistic interactions with other cytokines are critical in the interleukin-17 tuning effects. Macrophage inhibitory factor was described many years ago. Its comeback is based on properties of synoviocyte activation and proliferation. SUMMARY: Such findings are critical for a better understanding of response heterogeneity in patients treated with the cytokine inhibitors now on the market. New therapeutic approaches are been planned from these results.

Miossec P. · Department of Rheumatology, Hospital Edouard Herriot, Lyon, France. · Arthritis Rheum. · Pubmed #12632409 links to  free full text

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Miossec P. · Department of Immunology and Rheumatology, Hôpital Edouard Herriot, Lyon, France. · Cell Mol Biol (Noisy-le-grand). · Pubmed #11502074 No free full text.

Abstract: Anti-TNF-alpha therapy has shown clear efficacy in the treatment of rheumatoid arthritis (RA). Since some patients do not respond and the treatment is suspensive, combination therapy may be of interest. Other cytokines produced by monocytes such as IL-1, IL-12, IL-18 are also involved. The secretion of these cytokines is regulated by subsets of T-lymphocytes. Among these, IL-17 producing Th1 cells appear to contribute directly to the destructive process. Furthermore, this T-cell contribution enhances the action of monocyte derived proinflammatory cytokines. Using models of human RA synovium inflammation and bone resorption, ex vivo results suggest that combination therapy may be of interest. Acting on more than one cytokine may increase the percentage of responding RA patients as well as the degree of individual patient response.

Miossec P. · Department of Immunology, Hôpital Edouard Herriot, Lyon, France. · Curr Opin Rheumatol. · Pubmed #10803746 No free full text.

Abstract: T cells have been directly associated with rheumatoid arthritis (RA) because they represent the largest cell population infiltrating the synovium. Their direct contribution to disease and joint destruction has been more difficult to demonstrate. Locally, they interact with other blood-derived and resident cells. Some T cells may contribute to disease through the secretion of cytokines. Indeed, interleukin-17, a T-cell-specific cytokine, is produced by RA synovium and acts as a bone and cartilage destructive factor. In addition, it increases the production of proinflammatory cytokines by monocytes and further enhances their effects on matrix destruction. Once considered bystanders in RA, T cells can now be classified as aggressors through their direct and indirect contribution to destruction. In particular, a subset of Th1 T cells can aggravate the proinflammatory and destructive pattern associated with monocyte activation. Manipulation of this subset may control the destructive pattern. Such a result can be achieved when a switch can be induced from a destructive pattern to a protective one leading to repair.

Marotte H, Pallot-Prades B, Grange L, Gaudin P, Alexandre C, Miossec P. · Hospices Civils de Lyon-bioMérieux Research Unit on Rheumatoid Arthritis, Hopital Edouard Herriot, place d'Arsonval, 69437 Lyon Cedex 03, France. · Arthritis Res Ther. · Pubmed #17597527 links to  free full text

Abstract: The goal of the present study was to record changes in bone mineral density (BMD) and markers of bone turnover in patients with rheumatoid arthritis (RA) who were treated with methotrexate combined (or not combined) with infliximab. Included were 90 patients with RA who required anti-TNF-alpha therapy with infliximab because of persistent active disease despite treatment with methotrexate. The historical control group included 99 patients with RA who were treated with methotrexate at a time when anti-TNF-alpha treatment was not yet available. Lumbar and femoral neck BMD was measured using dual energy X-ray absorptiometry at baseline and 1 year later. Osteocalcin, C-terminal cross-linked telopeptide of type I collagen, parathyroid hormone and 25-hydroxycholecalciferol were measured in plasma at baseline and 1 year later. At 1 year BMD had decreased in the control group at spine (P < 0.01) and femoral neck (P < 0.001). In contrast, BMD at spine and femoral neck did not change after 1 year of infliximab treatment. At the same time point, no change in bone remodelling markers was observed. No association was observed between clinical response and changes in BMD, indicating that even those who did not respond clinically did not lose bone over a 1-year period. These data confirm the BMD decrease observed in RA patients treated with methotrexate alone. This bone loss was prevented by infliximab therapy. Importantly, this beneficial effect was also observed in apparent nonresponders.

Marotte H, Fontanges E, Bailly F, Zoulim F, Trepo C, Miossec P. · Clinical Immunology Unit, Department of Immunology, Edouard Herriot Hospital, France. · Rheumatology (Oxford). · Pubmed #16720634 links to  free full text

Abstract: OBJECTIVE: The treatment of the rheumatological manifestations associated with hepatitis C virus (HCV) remains difficult. To examine the safety of anti-tumour necrosis factor-alpha treatment, nine patients having rheumatological manifestations associated with HCV were treated with etanercept 25 mg twice a week for 3 months. METHODS: Five patients had a positive viral load at study entry (Group I), four were negative (Group II). Clinical data recorded were: disease duration, painful and swollen joint count, patient global and physician global assessment, the number of 18 specified fibromyalgia tender points and the Health Assessment Questionnaire score. Laboratory studies included checking for the presence of cryoglobulinaemia and transaminase levels. Quantitative HCV viral RNA was performed by real-time polymerase chain reaction (PCR). RESULTS: At 3 months, no patient was found to have evidence of increased hepatic inflammation based on serial serum transaminase levels. In the five patients from Group I with detectable HCV RNA, no significant viral load increase was observed. No reactivation was observed in the four patients from Group II with undetectable HCV RNA. The effect on the clinical rheumatological manifestations was more heterogeneous but appears to be lower than that observed in rheumatoid arthritis. CONCLUSION: In this phase II open short-term study, etanercept appeared to be safe in patients with articular manifestations associated with HCV.

Miossec P. · Department of Immunology and Rheumatology, Hôpital Edouard Herriot and Immunogenomics and inflammation research unit EA 4130, University of Lyon; Lyon, France. · Microbes Infect. · Pubmed #19371791 No free full text.

Abstract: IL-17 was discovered in 1995/96 as a T cell derived cytokine with effects on inflammation and neutrophil activation. In 2006, the precise cell source of IL-17 was identified in the mouse, and these cells were named Th17 cells. They play a role in various human diseases associated with inflammation and destruction such as rheumatoid arthritis, psoriasis, Crohn's disease, multiple sclerosis, where IL-17 can be seen as a therapeutic target.

Miossec P. · Department of Immunology and Rheumatology, Immunogenomics and inflammation Unit, University of Lyon, Lyon Cedex, France. · Eur J Immunol. · Pubmed #19283719 No free full text.

Abstract: The discovery of IL-17 and of the Th17 pathway has been a step in the classification of human diseases. Th17 targeting appears rather straightforward in diseases associated with inflammation and matrix destruction, such as rheumatoid arthritis, psoriasis, and Crohn's disease. In other conditions where IL-17 is expressed and Th17 activated, it is unclear whether this is a primary or secondary event, making the use of Th17 inhibitors less obvious.

Zrioual S, Ecochard R, Tournadre A, Lenief V, Cazalis MA, Miossec P. · Department of Immunology and Rheumatology and Hospices Civils de Lyon-bioMerieux Mixed Research Unit, Hospital Edouard Herriot, Lyon, France. · J Immunol. · Pubmed #19234208 No free full text.

Abstract: IL-17A is implicated in rheumatoid arthritis (RA) pathogenesis; however, the contribution of IL-17F remains to be clarified. Using microarrays and gene-specific expression assays, we compared the regulatory effects of IL-17A and IL-17F alone or in combination with TNF-alpha on RA synoviocytes. IL-17A and IL-17F expression was studied in osteoarthritis and RA synovium by immunohistochemistry. The comparison between the IL-17A and IL-17F stimulatory effect on RA synoviocytes was assessed at the protein level by ELISA and at the mRNA level by microarrays and real-time RT-PCR. TNFRII expression was studied by real-time RT-PCR and immunofluorescence, and neutralizing Ab was used to analyze its contribution to CCL20 secretion. IL-17A and IL-17F were detected in plasma cell-like cells from RA but not osteoarthritis synovium. In microarrays, IL-17A and IL-17F alone had similar regulatory effects, IL-17F being quantitatively less active. Both cytokines induced a similar expression pattern in the presence of TNF-alpha. Based on a cooperation index, 130 and 203 genes were synergistically induced by IL-17A or IL-17F plus TNF-alpha, respectively. Among these, the new target genes CXCR4, LPL, and IL-32 were validated by real-time RT-PCR. IL-17A and IL-17F up-regulated TNFRII expression, but had no effects on TNFRI, IL-17RA or IL-17RC. TNFRII blockade inhibited the synergistic induction of CCL20 by IL-17A or IL-17F and TNF-alpha. IL-17A and IL-17F are both expressed in RA synovium. In the presence of TNF-alpha, they induced a similar expression pattern in RA synoviocytes. Accordingly, IL-17F appears as a target in Th17-mediated diseases such as RA.

Mathieux R, Marotte H, Battistini L, Sarrazin A, Berthier M, Miossec P. · Clinical Immunology Unit, Department of Immunology, Hôpital Edouard Herriot, Lyon, France. · Ann Rheum Dis. · Pubmed #19015209 No free full text.

Abstract: AIM: The goal of occupational therapy (OT) is to facilitate adjustments to lifestyle and to prevent function loss. This study evaluated the effects of an early OT programme in early rheumatoid arthritis (RA). METHODS: We conducted a randomised, blind, controlled trial enrolling 60 patients with early RA, divided into 2 groups. At baseline, group 1 received the full information programme and group 2 received no information. In an extension phase, patients in group 2 received the full information programme at 3 months and were assessed at 6 months. The main outcomes were grip strength of hands (as objective assessment) and Health Assessment Questionnaire (HAQ) score (as subjective assessment). RESULTS: At 3 months, grip strength of the dominant and non-dominant hands increased more in group 1 than in group 2 (p = 0.021 and 0.047 respectively). HAQ score decreased more in group 1 than in group 2 (p<0.001). In the extension phase, changes in grip strength and HAQ score in group 2 were similar to those seen in group 1 between baseline and 3 months. CONCLUSIONS: This study comparing two schedules of OT programme showed that an early extended information programme improved hand function in patients with early RA.

Marotte H, Gineyts E, Miossec P, Delmas PD. · Hospices Civils de Lyon-bioMérieux Research Unit on Rheumatoid Arthritis, Hospital Edouard Herriot, Lyon, France. · Ann Rheum Dis. · Pubmed #18713784 No free full text.

Abstract: BACKGROUND: Defining the remission criteria of rheumatoid arthritis (RA) remains a critical issue. Markers of synovium activity, urinary glucosyl-galactosyl-pyridinoline (Glc-Gal-PYD) and of cartilage destruction, urinary C-terminal crosslinking telopeptide of type II collagen (CTX-II) have been shown to reflect disease activity and joint damage progression in RA. METHODS: The prospective study cohort comprised 66 RA patients treated with infliximab and methotrexate and 76 healthy controls. Measurements of urinary Glc-Gal-PYD and CTX-II were performed at baseline and at 1 year of infliximab therapy. RESULTS: At baseline, urinary Glc-Gal-PYD and CTX-II levels were increased in patients with RA and correlated with modified Sharp scores and progression of joint damage. Patients with more progressive joint destruction had higher Glc-Gly-PYD and CTX-II baseline levels. CONCLUSION: These markers reflected bone erosion evolution and might be useful for treatment monitoring and evaluation of RA. Markers remained high even in clinical responders after infliximab, suggesting persistence of synovitis.

Trocmé C, Marotte H, Baillet A, Pallot-Prades B, Garin J, Grange L, Miossec P, Tebib J, Berger F, Nissen MJ, Juvin R, Morel F, Gaudin P. · GREPI CNRS UMR 5525, INSERM IFR 130, Université J Fourier, Grenoble, France. · Ann Rheum Dis. · Pubmed #18664547 No free full text.

Abstract: OBJECTIVES: The use of biologicals such as infliximab has dramatically improved the treatment of rheumatoid arthritis (RA). However, factors predictive of therapeutic response need to be identified. A proteomic study was performed prior to infliximab therapy to identify a panel of candidate protein biomarkers of RA predictive of treatment response. METHODS: Plasma profiles of 60 patients with RA (28 non-responders (as defined by the American College of Rheumatology 20% improvement criteria (ACR20)) negative and 32 responders (ACR70 positive) to infliximab) were studied by surface enhanced laser desorption/ionisation time-of-flight mass spectrometry (SELDI-TOF MS) technology on two types of arrays, an anion exchange array (SAX2) and a nickel affinity array (IMAC3-Ni). Biomarker characterisation was carried out using classical biochemical methods (purification by ammonium sulfate precipitation or metal affinity chromatography) and identification by matrix assisted laser desorption/ionisation time-of-flight (MALDI-TOF) MS analysis. RESULTS: Two distinct protein profiles were observed on both arrays and several proteins were differentially expressed in both patient populations. Five proteins at 3.86, 7.77, 7.97, 8.14 and 74.07 kDa were overexpressed in the non-responder group, whereas one at 28 kDa was increased in the responder population (sensitivity>56%, specificity>77.5%). Moreover, combination of several biomarkers improved the sensitivity and specificity of the detection of patient response to over 97%. The 28 kDa protein was characterised as apolipoprotein A-I and the 7.77 kDa biomarker was identified as platelet factor 4. CONCLUSIONS: Six plasma biomarkers are characterised, enabling the detection of patient response to infliximab with high sensitivity and specificity. Apolipoprotein A-1 was predictive of a good response to infliximab, whereas platelet factor 4 was associated with non-responders.

Marotte H, Arnaud B, Diasparra J, Zrioual S, Miossec P. · Hôpital Edouard Hérriot, Lyon, France. · Arthritis Rheum. · Pubmed #18438841 links to  free full text

Abstract: OBJECTIVE: The tumor necrosis factor alpha (TNFalpha) -308A polymorphism has been associated with high production of TNFalpha and poor response to anti-TNFalpha therapy, but these associations remain controversial. The aim of this study was to explore the association between circulating TNFalpha bioactivity, the TNFalpha -308 polymorphism, and the clinical response to infliximab in patients with rheumatoid arthritis (RA). METHODS: One hundred ninety-eight patients with RA were treated with infliximab and methotrexate. Responses at 6 months according to the American College of Rheumatology (ACR) preliminary criteria for improvement in RA were recorded. Genotyping for the TNFalpha -308 polymorphism was performed by enzyme-linked oligosorbent assay. Circulating TNFalpha bioactivity was evaluated in 50 patients with RA by assessing the production of interleukin-6 (IL-6) in synoviocytes induced by a small amount of TNFalpha plus plasma. IL-6 production in 48-hour supernatants and the levels of TNFalpha protein and IL-6 were measured by enzyme-linked immunosorbent assay. RESULTS: The TNFalpha -308 polymorphism was not associated with the ACR response to infliximab. The level of circulating TNFalpha bioactivity was higher in patients with the TNFalpha -308 A/A or A/G genotype than that in patients with the G/G genotype (median 50.0 ng/ml [interquartile range (IQR) 31.5-62.0] versus 33.0 ng/ml [IQR 16.5-47.5]; P < 0.02). However, no difference was observed for the TNFalpha protein level according to genotype (median 0.62 pg/ml [IQR 0.00-8.85] for G/G versus 3.35 pg/ml [IQR 1.55-4.63] for A/A or A/G; P not significant). The level of circulating TNFalpha bioactivity was higher in good responders (> or =50% improvement) than in poor responders (< or =20% improvement) (median 45.0 ng/ml [IQR 21.0-59.0] versus 28.0 ng/ml [IQR 14.0-39.0]; P = 0.05). However, the level of TNFalpha protein was similar in both groups. CONCLUSION: The level of functional circulating TNFalpha is partially genetically determined and is predictive of the clinical response to infliximab. Nonresponders to anti-TNFalpha therapy are likely to have a disease that is not primarily driven by TNFalpha.

Zrioual S, Toh ML, Tournadre A, Zhou Y, Cazalis MA, Pachot A, Miossec V, Miossec P. · Department of Immunology and Rheumatology, Mixed Unit Civil Hospital of Lyon-Biomerieux, Lyon, France. · J Immunol. · Pubmed #18097068 links to  free full text

Abstract: IL-17A is a cytokine secreted by the newly described Th17 cells implicated in rheumatoid arthritis (RA). Less is known about its receptors in synoviocytes. IL-17RA and IL-17RC were found to be overexpressed in RA peripheral whole blood and their expression was detected locally in RA synovium. In vitro, IL-17A synergized with TNF-alpha to induce IL-6, IL-8, CCL-20, and matrix metalloproteinase-3. Using microarrays, a specific up-regulation of Glu-Leu-Arg+ CXC chemokines was observed in IL-17A-treated synoviocytes. Using both posttranslational inhibitions by silencing interfering RNA and extracellular blockade by specific inhibitors, we showed that both IL-17RA and IL-17RC are implicated in IL-17A-induced IL-6 secretion, whereas in the presence of TNF-alpha, the inhibition of both receptors was needed to down-regulate IL-17A-induced IL-6 and CCL-20 secretion. Thus, IL-17A-induced IL-6, IL-8, and CCL20 secretion was dependent on both IL-17RA and IL-17RC, which are overexpressed in RA patients. IL-17A-induced pathogenic effects may be modulated by IL-17RA and/or IL-17RC antagonism.

Pachot A, Arnaud B, Marrote H, Cazalis MA, Diasparra J, Gouraud A, Mougin B, Miossec P. · Joint Unit, Hospices Civils de Lyon-bioMérieux, Hôpital Edouard Herriot, Hôpital Edouard Herriot, 5 place d'Arsonval, Lyon, France. · J Rheumatol. · Pubmed #17896807 No free full text.

Abstract: OBJECTIVE: It has been suggested that patients with rheumatoid arthritis (RA) with abundant tumor necrosis factor-alpha (TNF-alpha) are more likely to respond to TNF-alpha inhibitors. We measured expression of TNF-alpha mRNA in peripheral blood of RA patients undergoing infliximab treatment in order to test its predictive value for treatment response. METHODS: Forty-four RA patients showing persistent disease activity and 27 healthy controls were studied. Peripheral blood TNF-alpha mRNA levels were measured before and 4 hours after the first infliximab infusion and at Week 22 using quantitative RT-PCR. Results were correlated to the treatment response at Week 22 in the whole RA cohort and a subset of patients showing high TNF-alpha mRNA levels at baseline. RESULTS: At baseline and at Week 22, TNF-alpha mRNA expression in RA patients was significantly increased compared to healthy controls. At both timepoints, no significant difference was observed between responders and nonresponders. Compared to baseline, infliximab treatment induced a decrease in TNF-alpha mRNA level at 4 hours and at Week 22, although this effect was significant only in patients with high TNF-alpha mRNA expression at baseline. Such variation compared to baseline was similar in responders and nonresponders. CONCLUSION: Peripheral blood TNF-alpha mRNA expression is increased in RA, but its reduction with anti-TNF treatment is not associated with treatment response.

Pachot A, Barbalat V, Marotte H, Diasparra J, Gouraud A, Mougin B, Miossec P. · Hospices Civils de Lyon, BioMérieux Joint Unit, Hôpital Edouard Herriot, 5, Place d'Arsonval, 69003, Lyon, France. · J Immunol Methods. · Pubmed #17884081 No free full text.

Abstract: Genomic DNA extraction for genotyping analysis is performed from blood samples and is time consuming. We describe a more rapid DNA extraction method, "DBS-miniMAG", that combines filter paper dried blood spots (DBS) with the NucliSens miniMAG semi-automated instrument (bioMérieux). To assess the performance of this method, a post-PCR HLA-DR shared epitope (SE) oligotyping assay was used as a read-out in a cohort of 72 arthritis patients. This new method was compared to the standard manual DBS extraction protocol using FTA reagents (Whatmann Bio-Science), and to a reference phenol-chloroform-based method using EDTA whole blood samples. Higher yield of PCR amplicons was observed with DNA extracts obtained using "DBS-miniMAG" method. The intra- and inter-assay variability of the "DBS-miniMAG" method was similar to that obtained with "DBS-FTA" washing process. Concerning the HLA-DR SE genotyping, "DBS-miniMAG" and "DBS-FTA" methods gave 100% concordance compared to the reference phenol-chloroform method. More importantly, the hands-on time and the turnaround time for "DBS-miniMAG" were both two-times shorter than for "DBS-FTA" protocol. Therefore, the "DBS-miniMAG" combination could facilitate polymorphism analysis in routine clinical practice and the creation of large DNA banks using very small amounts of blood.

Cimaz R, Cazalis MA, Reynaud C, Gerloni V, Zulian F, Biggioggero M, Martini G, Pontikaki I, Fantini F, Mougin B, Miossec P. · Unité Mixte Hospices Civils de Lyon-BioMérieux, Lyon, France. · Ann Rheum Dis. · Pubmed #17324969 No free full text.

Abstract: OBJECTIVE: To investigate the genetic contribution of cytokine gene polymorphisms (interleukin 1 (IL1) and tumour necrosis factor alpha (TNFalpha)) on disease phenotype and on response to TNF-blocking agents in a population of patients with juvenile idiopathic arthritis (JIA). METHODS: A cohort of 107 consecutive patients with JIA who were receiving treatment with anti-TNF agents was enrolled in this study. Analysis of genetic polymorphisms for IL1B +3954, IL1RA +2018, TNFalpha -238 and TNFalpha -308 was performed by enzyme-linked oligo sorbent assay, and compared with those obtained from 630 healthy Caucasians and 263 adult patients with rheumatoid arthritis. Relevant demographic, clinical and laboratory data were collected from clinical charts and entered into a customised database, and chi(2) analysis was performed to compare cytokine polymorphisms with disease type according to the International League of Associations for Rheumatology criteria, presence of uveitis, rheumatoid factor and anti-nuclear antibody positivity, erosive disease, frequency of adverse effects to anti-TNF and clinical response after 3 months. RESULTS: The T/T genotype of the IL1B +3954 polymorphism was absent in patients with JIA and present in 5% of controls (p = 0.015). No significant correlation was found between the studied polymorphisms and clinical or laboratory variables considered. Clinical response to TNF inhibitors at 3 months was not associated with the genetic polymorphisms considered. CONCLUSION: In our cohort, the absence of the rare IL1B +3954 gene polymorphism was associated with JIA, but without specificity to particular disease phenotypes. The TNF and IL1 gene polymorphism studied did not seem to be associated with response to anti-TNF treatment.

Kazkaz L, Marotte H, Hamwi M, Angélique Cazalis M, Roy P, Mougin B, Miossec P. · Clinical Immunology Unit, Departments of Immunology and Rheumatology, Hôpital Edouard Hérriot, 69437 Lyon Cedex 03, France. · Ann Rheum Dis. · Pubmed #17068065 No free full text.

Abstract: OBJECTIVE: To investigate whether ethnic differences exist in the effect of the shared epitope and selected cytokine gene polymorphisms on the susceptibility and severity of rheumatoid arthritis in Syria (Damascus) and France (Rhône-Alpes area). METHODS: 156 patients with rheumatoid arthritis and 120 healthy controls from Syria were compared with 512 patients with rheumatoid arthritis and 471 healthy controls from France. Shared epitope status, cytokine gene polymorphisms interleukin (IL)-1B +3954, IL-1RN +2018 and tumour necrosis factor alpha promoter (-238 and -308) were analysed by enzyme-linked oligosorbent assay. Joint destruction was defined by a right wrist Larsen score > or =2. Odds ratios (ORs) were calculated. RESULTS: In both countries, a dose effect was observed between the shared epitope copy number and rheumatoid arthritis (Syria: OR 1 v 0 copies = 1.6, p = NS; OR 2 v 0 = 15.3, p<0.01; and France: OR 1 v 0 = 2.3, p<0.001; OR 2 v 0 = 7.2, p<0.001). A dose effect was also observed between the shared epitope copy number and joint destruction in Syria (OR 1 v 0 = 2.2, p = NS; OR 2 v 0 = 9.9, p<0.01) and France (OR 1 v 0 = 1.8, p<0.01; OR 2 v 0 = 4.8, p = 0.001). The dose effect of the shared epitope was greater in Syria than in France. Only the -238 tumour necrosis factor alpha polymorphism was associated with joint destruction in the Syrian population (p<0.05). However, after adjustment for age, sex, disease duration and rheumatoid factor for severity, this association disappeared. CONCLUSION: The frequency of the shared epitope was increased in the French population with rheumatoid arthritis and in controls, but the association between the shared epitope and joint destruction was more pronounced in the Syrian population, with an OR of almost 10 for the homozygotes.

Marotte H, Tournoud M, Cazalis MA, Mougin B, Roy P, Miossec P. · 1Department of Immunology and Rheumatology, Unité Mixte Hospices Civils de Lyon - BioMérieux, Hôpital Edouard Herriot, Lyon Cedex, France. · Genes Immun. · Pubmed #16826237 No free full text.

Abstract: To analyse the association between individual HLA-DRB1 locus genotypes and rheumatoid arthritis (RA) susceptibility, taking in account the multiallelic nature of the shared epitope (SE). In total, 538 patients and 536 controls were genotyped for 12 alleles of the HLA-DRB1 locus. A Bayesian partition model and multivariate logistic models were used to assess the role of the SE and of its individual components. The SE was associated with RA susceptibility (odds ratio (OR) 2 versus 0 SE copy=9.99 (95 CI 4.69-15.30) and OR 1 versus 0 SE copy=3.16 (95% CI 2.42-4.12)). The Bayesian partition model supplied a permutation of the HLA-DRBA locus alleles ordered by increasing disease risk. Alleles associated with highest risks are those that code for the SE. The individual OR estimations for the HLA-DRB1 locus genotypes went from OR=1.00 (95% CI 1.00-1.25) for the less associated genotype to OR=21.40 (95% CI 8.02-65.79) for the most associated one. In conclusion, the allele order risk and the OR estimations for individual genotypes of the HLA-DRB1 locus were consistent with the SE theory. Using an exploratory statistical method without a priori hypothesis, our study allowed a detailed analysis of the multiallelic nature of the SE.

Toh ML, Marotte H, Blond JL, Jhumka U, Eljaafari A, Mougin B, Miossec P. · Unité Mixte Hospices Civils de Lyon-BioMérieux, Hôpital Edouard Herriot, Lyon, France. · Arthritis Rheum. · Pubmed #16802346 links to  free full text

Abstract: OBJECTIVE: Synoviolin is a novel E3 ubiquitin ligase that has been implicated in the pathogenesis of rheumatoid arthritis (RA). The purpose of this study was to examine the expression and regulation of synoviolin by tumor necrosis factor alpha (TNFalpha), both in vivo and in vitro. METHODS: A total of 54 RA patients and 23 healthy control subjects were analyzed before, 4 hours after the first infusion, and at week 22 of infliximab treatment. Clinical response was assessed by the American College of Rheumatology criteria for 20% improvement and the Disease Activity Score in 28 joints (DAS28) at 6 months. Synoviolin messenger RNA expression was measured by real-time reverse transcription-polymerase chain reaction in peripheral blood (PB) and fibroblast-like synoviocytes (FLS) and with and without TNFalpha or interleukin-1beta (IL-1beta) stimulation. RESULTS: Synoviolin expression was increased in whole PB obtained from RA patients as compared with that from healthy controls and was significantly reduced early and late after infliximab treatment in responders, but in not nonresponders. Reduction in synoviolin expression was associated with reduced levels of markers of disease activity, including C-reactive protein levels. Nonresponders to infliximab therapy had significantly higher synoviolin expression at baseline as compared with responders, and this elevation persisted despite infliximab therapy. PB CD14+ monocytes expressed increased synoviolin levels compared with CD3+ lymphocytes, and TNFalpha or IL-1beta induced a further increase in expression in CD3+ cells. TNFalpha or IL-1beta induced sustained synoviolin expression in RA FLS. CONCLUSION: Elevated PB levels of synoviolin were identified in circulating PB mononuclear cells and were associated with nonresponse to infliximab treatment. Sustained up-regulation of synoviolin by IL-lbeta and TNFalpha may contribute to prolonged survival of immune cells and dysregulated FLS proliferation, leading to RA chronicity.