Rheumatoid Arthritis: Boissier MC

Boissier MC, Assier E, Falgarone G, Bessis N. · No affiliation provided · Joint Bone Spine. · Pubmed #18571969 No free full text.

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Falgarone G, Duclos M, Boissier MC. · No affiliation provided · Joint Bone Spine. · Pubmed #18092377 No free full text.

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Boissier MC, Bessis N, Falgarone G. · No affiliation provided · Joint Bone Spine. · Pubmed #12184428 No free full text.

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Boissier MC, Assier E, Biton J, Denys A, Falgarone G, Bessis N. · Inserm ERI18, Bobigny, France. · Joint Bone Spine. · Pubmed #19028128 No free full text.

Abstract: Modulation of the T-cell response depends chiefly on regulatory T cells (Treg), which express CD4 and CD25. Some Treg cells are present naturally, whereas others are induced in response to antigens. The immunomodulating effects of Treg cells are mediated by membrane molecules (e.g., CTLA4, GITR, and OX40) and cytokines. IL-35 seems to be a crucial mediator, although IL-10 and TGFbeta are also important. The role for Treg cells in rheumatoid arthritis (RA) has been established in both patients and animal models. Treg function is deficient in RA, whereas Treg counts vary. Treg counts increase in patients who are responding to TNFalpha antagonist therapy. Among current hypotheses, Treg expansion or transfer may hold promise for the treatment of RA.

Lormeau C, Falgarone G, Roulot D, Boissier MC. · Rheumatology Department and INSERM ERI-18, Avicenne Teaching Hospital (AH-HP) and Paris 13 University, 125, rue de Stalingrad, 93009 Bobigny, France. · Joint Bone Spine. · Pubmed #17056293 No free full text.

Abstract: The many rheumatologic manifestations associated with chronic hepatitis C virus (HCV) infection include arthralgia, myalgia, arthritis, vasculitis, and sicca syndrome. Arthralgia is the most common extrahepatic manifestation and may indicate mixed cryoglobulinemia or an adverse reaction to interferon therapy. HCV arthritis unrelated to cryoglobulinemia is far less common but constitutes an independent entity. The picture may mimic rheumatoid arthritis (RA), particularly as rheumatoid factor is present in 50-80% of cases. Tests are usually negative for antibodies to cyclic citrullinated peptides (anti-CCP), which may help to differentiate the two conditions. The management of HCV arthritis is empirical and poorly standardized. Although low-dose glucocorticoid therapy, hydroxychloroquine, and methotrexate have been used successfully in several patients, little is known about their hepatic safety profile. Arthritis associated with cryoglobulinemia usually responds to antiviral treatment. Sicca syndrome is common in patients with chronic HCV infection and shares similarities with primary Sjögren syndrome, suggesting that HCV infection may deserve to be included among the causes of secondary Sjögren syndrome. HCV-associated vasculitis is usually related to cryoglobulinemia, although a few cases of polyarteritis nodosa-like disease affecting the medium-sized vessels have been reported. Other conditions reported in patients with chronic HCV infection include fibromyalgia, systemic lupus erythematosus (SLE), antiphospholipid syndrome, and osteosclerosis.

Bessis N, Boissier MC. · Service de rhumatologie, UPRES EA-3408, immunologie, université Paris 13, CHU d'Avicenne, AP-HP, 74, rue Marcel-Cachin, Bobigny cedex, France. · Joint Bone Spine. · Pubmed #16226478 No free full text.

Abstract: Gene therapy seeks either to supply a missing or dysfunctional gene or to ensure continuous long-lasting production of a therapeutic protein. Rheumatoid arthritis is a candidate for gene therapy, as the mechanisms leading to joint inflammation and destruction have been partly elucidated. Nevertheless, several crucial questions need to be addressed. Knowledge of the underlying pathophysiological mechanisms is needed to guide selection of the candidate gene. In the light of current data, TNF and IL-1 antagonists are generating interest. A choice must be made between a viral vector (adenovirus, retrovirus, adeno-associated virus) and a nonviral vector (naked DNA, administered by electrotransfer or in liposomes). Finally, the relative merits of intraarticular and systemic administration need to be considered. Safety is a primary concern. The transgene and/or vector may induce adverse effects. For instance, a transgene inserted within the host genome (when a retroviral vector is used) may induce a mutation. A number of vectors and transgenes induce immune responses. Numerous studies are ongoing to investigate the safety and efficacy of gene therapy strategies in experimental models of rheumatoid arthritis. These studies will have to be completed before further clinical trials of gene therapy in rheumatoid arthritis are considered.

Falgarone G, Jaen O, Boissier MC. · Rheumatology Department, UPRES EA-3408, Hôpital Avicenne (AP-HP), Paris 13 University, Bobigny, France. · Joint Bone Spine. · Pubmed #15681243 No free full text.

Abstract: Innate immunity is the first line of defense against pathogenic microorganisms (bacteria, viruses, fungi, and parasites). After a long period of neglect, innate immunity is again recognized as a key mechanism not only in preventing invasion of the body by microorganisms, but also in contributing to the pathogenesis of autoimmune and inflammatory diseases by deviating the immune response or promoting the emergence of a regulatory response. The many factors involved in innate immunity often act in parallel or in alternation to generate adaptive immune responses. Innate immune responses are specific for groups of molecules or macromolecules found in components of microorganisms, usually the cell wall. The cellular and protein effectors of innate immunity are found in the rheumatoid synovium, and an increasing body of evidence indicates that they are directly involved in joint inflammation and in destruction of the joint cartilage and bone. In addition, they may have regulatory effects on inflammation and immunity. Whether innate immune mechanisms are causes or consequences of inflammation, and whether they regulate or amplify adaptive immune responses, they constitute a target of choice for new antiinflammatory and immunoregulating treatment strategies.

Rat AC, Boissier MC. · Rheumatology Department, Avicenne Teaching Hospital (AP-HP), 125, route de Stalingrad, CHU Avicenne, 93009 Bobigny, France. · Joint Bone Spine. · Pubmed #15589432 No free full text.

Abstract: Rheumatoid arthritis (RA) causes disability, deformities, progressive radiological joint damage often with a need for joint replacement surgery, premature death, and alterations in quality of life. The economic burden created by RA is enormous. Direct costs per patient have been estimated at 1812-11,792 Euro annually and indirect costs at 1260-37,994 Euro annually. These mean values are approximations, as variations occur across countries, healthcare system organizations, and geographic locations. Direct costs account for one-fourth to slightly over a half of the total cost. Costs associated with inpatient care contribute up to 75% of direct costs, as compared to only about 20% for medications, although wide variations occur in costs related to drug monitoring and side-effect management. Physician visits account for about 20% of direct costs. As compared to indirect health costs for individuals from the general population, those for RA patients are increasing at a rapid rate. Indirect costs account for 80% of the excess cost related to RA. Cost estimates may change over time and show huge variations across individuals, with a small minority of patients accounting for most of the costs. Disability as measured by the Health Assessment Questionnaire (HAQ) has a major impact on costs. Early effective treatment may not only postpone and slow disease progression, thereby improving quality of life, but also decrease costs by preserving productivity and reducing the need for surgery, admission to acute-care and extended-care hospitals, and social service utilization. Data are beginning to accumulate on the excess costs associated with biotherapies and other new second-line drugs. They indicate acceptable excess costs relative to the additional medical benefits and to the cost-effectiveness of other healthcare programs. Nevertheless, the threshold that defines an acceptable excess cost is arbitrary and varies with local economic conditions.

Boissier MC, Bessis N. · Service de Rhumatologie, Hopital Avicenne, Bobigny, France. · Reumatismo. · Pubmed #15201941 links to  free full text

Abstract: Rheumatoid arthritis (RA) is a severe autoimmune systemic disease. Chronic synovial inflammation results in destruction of the joints. No conventional treatment is efficient in RA. Gene therapy of RA targets mainly the players of inflammation or articular destruction: TNF-alpha or IL-1 blocking agents (such as anti-TNF-alpha monoclonal antibodies, soluble TNF-alpha receptor, type II soluble receptor of IL-1, IL-1 receptor antagonist), anti-inflammatory cytokines (such as IL-4, IL-10, IL-1), growth factors. In this polyarticular disease, the vector expressing the therapeutic protein can be administered as a local (intra articular injection) or a systemic treatment (extra articular injection). All the main vectors has been used in experimental models, including the more recent lentivirus and adeno-associated virus. Ex vivo gene transfer was done with synovial cells, fibroblasts, T cells, dendritic cells, and different cells from xenogenic origin. In vivo gene therapy is simpler, although less controlled method. Clinical trials in human RA has started with ex vivo retrovirus expressing IL-1 receptor antagonist and have demonstrated the feasibility of the strategy of gene therapy. The best target remains to be determined and extensive researches have to be conducted in pre-clinical studies.

Rat AC, Henegariu V, Boissier MC. · Rheumatology Department, CHU Avicenne (AP-HP), Bobigny cedex, France · Joint Bone Spine. · Pubmed #15182789 No free full text.

Abstract: OBJECTIVE: Few recommendations have been issued about the management of rheumatoid arthritis (RA), which varies widely across physicians. The primary care physician (PCP) plays a unique role as the first physician to evaluate the patient. The objective of this study was to evaluate the place of PCPs in the management of RA. METHODS: Medline was searched for articles reporting management of rheumatoid arthritis in primary care practice. RESULTS: Currently, the goal of initiating a disease modifying anti-rheumatic drug (DMARD) early is unrealistic for numerous patients. Agreement between PCPs and rheumatologists about the diagnosis of RA is only passable, but PCPs tend to overdiagnose RA. Median time from symptom onset to second-line treatment was 19 months and the best predictive factor for a longer lag time before DMARD prescription was the time from symptom onset to the first rheumatologist visit. Moreover, DMARDs are only rarely prescribed by PCPs. Some data suggest that the impact of rheumatologists care is positive on outcomes but it has to be confirmed by longitudinal, randomized studies, with valid outcomes and diagnosis criteria. Recognition of the need for timely referral is an important goal in the teaching of students and generalists. Moreover, the nature of management differences between rheumatologists and PCPs has to be explored. We should also think how to create a better coordination. This starts by knowing what are the needs of the PCP (e.g. education, access to phone advice or rapid consultation) and by defining common plan if the care should be shared. CONCLUSION: Several healthcare professionals, among whom the PCP plays a pivotal role, should share the management of RA. PCPs and rheumatologists should be encouraged to work together on optimizing the management of patients with RA.

Saidenberg-Kermanac'h N, Cohen-Solal M, Bessis N, De Vernejoul MC, Boissier MC. · Groupe de recherche en immunopathologie et immunointervention (UPRES EA-3408) et service de rhumatologie (Hôpital Avicenne, AH-AP), Université Paris 13, UFR Léonard de Vinci, Bobigny, France. · Joint Bone Spine. · Pubmed #14769514 No free full text.

Abstract: Osteoprotegerin (OPG), a member of the TNF-receptor family expressed by osteoblasts, has documented effects on the regulation of bone metabolism. OPG inhibits bone resorption and binds with strong affinity to its ligand RANKL, thereby preventing RANKL from binding to its receptor RANK. This system is regulated by calcium-modifying hormones. OPG may also be pivotal in modulating the immune system. RANKL-deficient mice exhibit both severe immunological abnormalities and osteopetrosis, and activated T cells express RANKL mRNA. RANKL secretion by activated T cells may induce osteoclastogenesis via a mechanism enhanced by several cytokines (TNF-alpha, IL-1, and IL-17) that promote both inflammation and bone resorption. Conversely, this mechanism is inhibited by OPG, IL-4, and IL-10, which have antiinflammatory effects and inhibit osteoclast formation. Activated T cells in the rheumatoid synovium express RANKL. Synoviocytes can differentiate to osteoclast-like cells under specific conditions, particularly when they are cultured with M-CSF and RANKL. Thus, the bony erosions seen in RA may result from RANKL/RANK system activation by activated T cells. This raises the possibility that OPG therapy to block this mechanism might prove beneficial in patients with RA.

Clavel G, Bessis N, Boissier MC. · Service de Rhumatologie, CHU Nord, Place Victor Pauchet, 80054 Amiens cedex 1, France. · Joint Bone Spine. · Pubmed #14563458 No free full text.

Abstract: Angiogenesis is central to the development and perpetuation of rheumatoid synovitis. Vascular endothelial growth factor (VEGF), the main mediator of angiogenesis, is found in the synovial fluid and serum of patients with rheumatoid arthritis (RA), and its expression is correlated with disease severity. Compelling evidence that VEGF is involved in synovitis has been obtained from experimental models of RA. In particular, VEGF inhibition by synthetic compounds (e.g. TNP-470) or by naturally occurring factors (e.g., the soluble VEGF receptor) produce therapeutic effects. Angiopoietin-1, a recently discovered growth factor specific for neovascularization, is expressed within the rheumatoid synovium and may be stimulated by TNF-alpha. Other compounds, including integrins, fibroblast growth factor, and proinflammatory cytokines contribute to joint angiogenesis and, therefore, to the development of rheumatoid synovitis. Assessing vascularity may prove useful for evaluating or even predicting bone destruction. Furthermore, inhibition of angiogenesis may prove useful as an adjunct to current anti-inflammatory treatments.

Bessis N, Doucet C, Cottard V, Douar AM, Firat H, Jorgensen C, Mezzina M, Boissier MC. · UPRES EA-3408 (Université Paris 13) and Service de Rhumatologie (CHU Avicenne, AP-HP), Bobigny, France. · J Gene Med. · Pubmed #12439850 No free full text.

Abstract: Rheumatoid arthritis (RA) is a severe autoimmune systemic disease. Chronic synovial inflammation results in destruction of the joints. No conventional treatment is efficient in RA. Gene therapy of RA targets mainly the players of inflammation or articular destruction: TNF-alpha or IL-1 blocking agents (such as anti-TNF-alpha monoclonal antibodies, soluble TNF-alpha receptor, type II soluble receptor of IL-1, IL-1 receptor antagonist), antiinflammatory cytokines (such as IL-4, IL-10, IL-1), and growth factors. In this polyarticular disease, the vector expressing the therapeutic protein can be administered as a local (intra-articular injection) or a systemic treatment (extra-articular injection). All the main vectors have been used in experimental models, including the more recent lentivirus and adeno-associated virus. Ex vivo gene transfer was performed with synovial cells, fibroblasts, T cells, dendritic cells, and different cells from xenogeneic origin. In vivo gene therapy is simpler, although a less controlled method. Clinical trials in human RA have started with ex vivo retrovirus-expressing IL-1 receptor antagonists and have demonstrated the feasibility of the strategy of gene therapy. The best target remains to be determined and extensive research has to be conducted in preclinical studies.

Maravic M, Daurès JP, Boissier MC. · Service de rhumatologie, UPRES EA-3408, CHU Avicenne (AP-HP), Université Paris, Bobigny, France. · Joint Bone Spine. · Pubmed #12102273 No free full text.

Abstract: The objective of this study was to review evaluations of current clinical practice among rheumatologists managing patients with rheumatoid arthritis. We reviewed articles retrieved through PubMed using the search terms 'rheumatoid arthritis', 'clinical practice', 'physician's practice patterns', and 'health service research'. We set the time limits at January 1990 and May 2001. Marked variability was found in initial tests, initial treatment, changes in clinical status by number of visits, second-line treatment prescription and monitoring, use of monitoring criteria, clinical outcomes by type of health care insurance, and management of a fictional case. The data were obtained by survey in some studies and by prospective collection in others. This literature review indicates a need for developing recommendations for the management of rheumatoid arthritis.

Saidenberg Kermanac'h N, Bessis N, Cohen-Solal M, De Vernejoul MC, Boissier MC. · Groupe de Recherches en Immunopathologie et Immunointervention, UPRES EA-3408, Formation associée Claud Bernard and Service de Rhumatologie, Hôpital Avicenne, AH-AP, Université Paris 13, UFR Léonard-de Vinci, Bobigny, France. · Eur Cytokine Netw. · Pubmed #12101070 No free full text.

Abstract: RANK, RANKL, and OPG have well established regulatory effects on bone metabolism. RANK is expressed at very high levels on osteoclastic precursors and on mature osteoclasts, and is required for differentiation and activation of the osteoclast. The ligand, RANKL binds to its receptor RANK to induce bone resorption. RANKL is a transmembrane protein expressed in various cells type and particularly on osteoblast and activated T cells. RANKL can be cleaved and the soluble form is active. Osteoprotegerin decoy receptor (OPG), a member of the TNF receptor family expressed by osteoblasts, strongly inhibits bone resorption by binding with high affinity to its ligand RANKL, thereby preventing RANKL from engaging its receptor RANK. This system is regulated by the calciotropic hormones. Conversely, the effects of RANKL, RANK, and OPG on inflammatory processes, most notably on the bone resorption associated with inflammation, remain to be defined. The RANK system seems to play a major role in modulating the immune system. Activated T cells express RANKL messenger RNA, and knock-out mice for RANKL acquire severe immunological abnormalities and osteopetrosis. RANKL secretion by activated T cells can induce osteoclastogenesis. These mechanisms are enhanced by cytokines such as TNF-alpha, IL-1, and IL-17, which promote both inflammation and bone resorption. Conversely, this system is blocked by OPG, IL-4, and IL-10, which inhibit both inflammation and osteoclastogenesis. These data may explain part of the abnormal phenomena in diseases such as rheumatoid arthritis characterized by both inflammation and destruction. Activated T cells within the rheumatoid synovium express RANKL. Synovial cells are capable of differentiating to osteoclast-like cells under some conditions, including culturing with M-CSF and RANKL. This suggests that the bone erosion seen in rheumatoid arthritis may result from RANKL/RANK system activation by activated T cells. This opens up the possibility that OPG may have therapeutic effects mediated by blockade of the RANKL/RANK system.

Bessis N, Boissier MC. · UPRES EA-3408 et Formation de Recherche en Immunopathologie et Immuno-Intervention Articulaires (Association Claude Bernard), Rheumatology Department (CHU Avicenne, AP-HP), UFR Léonard de Vinci Bobigny, Université Paris 13, France. · Joint Bone Spine. · Pubmed #11808983 No free full text.

Abstract: Among potential targets for nonspecific anti-inflammatory immunointervention, three pro-inflammatory interleukins (ILs) have recently been found to play a pivotal role in rheumatoid arthritis (RA). IL-15 has both chemoattractant and proinflammatory properties and may promote bone destruction. IL-17, a product of T lymphocytes, has proinflammatory effects and induces production of metalloproteinases such as MMP-1. IL-18 not only has proinflammatory, angiogenic, and chemoattractant effects but also promotes cartilage destruction. These cytokines are potential targets for specific or nonspecific anti-inflammatory therapy. Thus, blocking IL-15 by its receptor reduces the severity of experimental collagen-induced arthritis (CIA). In this model, IL-17 levels fall after administration of anti-inflammatory cytokines such as IL-4 or IL-13. Finally, monoclonal anti-IL-18 antibodies prevent streptococcal cell wall arthritis, and IL-18 binding protein, which is a naturally occurring IL-18 inhibitor, prevents CIA.

Guedes C, Dumont-Fischer D, Leichter-Nakache S, Boissier MC. · Rheumatology Department and Joint Immunopathology and Immunointervention Unit, Bobigny-Avicenne Teaching Hospital. · Rev Rhum Engl Ed. · Pubmed #10567978 No free full text.

Abstract: Rheumatoid arthritis reduces not only quality but also length of life. In the 14 main studies conducted since 1980, in a total of 13,424 patients, the mean standardized mortality ratio was 1.82 (range, 0.87-3) as compared to the population at large. Life expectancy was shortened by 5 to 10 years in most studies. The diversity of the methods used explains the discrepancies among results. Excess mortality may occur in only some subsets of patients. Both rheumatoid complications and an increase in nonspecific causes of death (e.g., infections) contribute to the excess mortality. Factors predictive of premature death are the same as those predictive of functional impairment. Many unknowns remain about the condition of rheumatoid arthritis patients at the end of their life.

Falgarone G, Semerano L, Rullé S, Boissier MC. · University of Paris 13, Li2P, EA4222, Bobigny, France. · Joint Bone Spine. · Pubmed #19535279 No free full text.

Abstract: The introduction of targeted treatments has radically changed the management of patients with rheumatoid arthritis (RA). Abatacept is among these new treatments emerging from recent insights into joint immunopathology. Abatacept blocks the interaction between antigen-presenting cells and T-cells, thereby diminishing T-cell activation and possibly improving overall cell regulation. In RA patients, abatacept is effective in decreasing the arthritis, pain, disability, fatigue, and radiological joint damage. Abatacept provides lasting remissions or low levels of disease activity and therefore constitutes a valuable addition to the current therapeutic armamentarium for RA, which is hoped to make a full remission an attainable goal in the overall population of RA patients.

Jaen O, Petit-Teixeira E, Kirsten H, Ahnert P, Semerano L, Pierlot C, Cornelis F, Boissier MC, Falgarone G, Anonymous00012. · EA-4222, University of Paris 13, Bobigny Cedex, Paris, France. · Arthritis Res Ther. · Pubmed #19134200 links to  free full text

Abstract: INTRODUCTION: The objective was to study the potential genetic contribution of Toll-like receptor (TLR) genes in rheumatoid arthritis (RA). TLRs bind to pathogen-associated molecular patterns, and TLR genes influence both proinflammatory cytokine production and autoimmune responses. Host-pathogen interactions are involved in RA physiopathology. METHODS: We tested SNPs of five TLR genes (TLR9, TLR2, TLR6, TLR1, and TLR4) in a cohort of 100 French families with RA. Genotypes were analyzed using the transmission disequilibrium test. As TLR2, TLR6, and TLR1 are located on chromosome 4, we determined the haplotype relative risk. Analyses were performed in subgroups defined by status for rheumatoid factor, anti-cyclic citrullinated peptide autoantibodies, and erosions. RESULTS: We found no disequilibrium in allele transmission for any of the SNPs of the five TLR genes. In subgroup analyses, no associations were detected linking TLR9, TLR2, or TLR9/TLR2 to rheumatoid factor, anti-cyclic citrullinated peptide autoantibodies, or erosions. Haplotype analysis of the polymorphisms showed no haplotype associations in any of the subgroups. CONCLUSIONS: We found no evidence of major effects of TLR gene polymorphisms in RA, although we tested different TLR phenotypes. Moreover, no associations were noted with autoantibody production or erosions.

Launois R, Payet S, Saidenberg-Kermanac'h N, Francesconi C, França LR, Boissier MC. · REES France, Paris, France. · Joint Bone Spine. · Pubmed #18951825 No free full text.

Abstract: OBJECTIVES: To estimate the budget impact implied by the introduction of rituximab after failure of one or more anti-TNFalpha therapies in the perspective of the French health care system. METHODS: A Markov model reproduced the course, over 4years, of patients treated either by infliximab, etanercept, adalimumab or RTX, after failure of one or more anti-TNFalpha therapies, in a multicentric study. A sensitivity analysis was developed to account for patients in 3rd and subsequent lines of treatment who are expected to consume more healthcare resources. RESULTS: When RTX is not used, total annual medical cost is euro16,555 per patient, euro13,206 of which are dedicated to drug acquisition. When RTX is the only treatment in use, these costs decrease respectively to euro11,444 and euro7469. Total savings per patient and per year is euro5000. Over 4 years, total savings for the targeted population reach euro118M. In the sensitivity analysis, the difference between H2 and H2-coeff 2 (20%) reaches euro5,400,000 in total direct costs during the first year of simulation. This difference decreases along the period, to reach euro2,400,000 the fourth year of simulation, and is due to the fact that rituximab acquisition costs are independent from the treatment line. CONCLUSION: If TNFalpha inhibitors were the only treatment available, the annual global cost of treatment would be euro16,555 per patient versus euro11,444 for patients treated exclusively with rituximab. RTX is expected to produce important savings (-31%) if used after failure of one or more TNFalpha therapies. This is mainly due to its lower drug acquisition cost. These savings could increase with the development of rituximab in earlier stages of treatment.

Jaen O, Rullé S, Bessis N, Zago A, Boissier MC, Falgarone G. · INSERM ERI18, Paris 13 University, AP-HP Rheumatology Department, Avicenne Hospital, Bobigny, France. · Immunology. · Pubmed #18754812 No free full text.

Abstract: Dendritic cells (DCs) mediate interactions between innate and specific immunity and may induce regulatory mechanisms. We investigated the effects of modulated DCs in mice with collagen-induced arthritis (CIA) and tested the responses of cells to induced naturally occurring regulatory T cells. DCs were stimulated or not with DNA or lipopolysaccharide (LPS) for 24 hr. DC maturation was assayed, and then modulated DCs were intraperitoneally injected on day 14 into DBA/1 mice to treat CIA. In addition to arthritis scores and type 2 collagen (CII) response, the induction of CD4(+) CD25(+) T cells was analysed by flow cytometry in peripheral blood and the expression of Foxp3, transforming growth factor (TGF)-beta, interleukin (IL)-10 and cytotoxic T-lymphocyte antigen (CTLA)-4 was quantified. Finally, the expression of indoleamine-2,3-dioxygenase (IDO) was assayed in DCs. In comparison with LPS-stimulated DCs, plasmid-stimulated DCs expressed lower levels of major histocompatibility complex (MHC) class II, CD40, CD80 and CD86 molecules and secreted less IL-12p70, interferon (IFN)-gamma, IL-10 and TNF-alpha, displaying a semi-mature phenotype. Compared with non-stimulated DCs, stimulated DCs improved arthritis scores when injected after immunization, without modifying the T helper type 1 (Th1)/Th2 balance of the immune response against collagen. Stimulated DCs induced markers for regulatory T cells (Foxp3, TGF-beta1 and CTLA-4) in vivo. Only LPS-stimulated DCs expressed IDO, which may explain their better therapeutic efficacy. Regulatory mechanisms were induced using DCs modulated by innate immunity stimulators. Innate immunity mechanisms do not require the presence of the disease-causing antigen, even in T- and B-cell specific diseases. Our results have implications for the treatment of rheumatoid arthritis, an autoimmune disease whose triggering antigen has not been identified, and substantially clarify the role of regulatory T cells in CIA.

Clavel G, Marchiol-Fournigault C, Renault G, Boissier MC, Fradelizi D, Bessis N. · INSERM ERI-18, Université Paris 13, France. · Ann Rheum Dis. · Pubmed #18218664 No free full text.

Abstract: OBJECTIVES: The evaluation of joints in arthritis using conventional ultrasonography is not really feasible in mice because of the small size of the animal. However, compared with classical analysis (clinical and histological examination) it is a non-invasive method that allows follow-up of the same animal throughout the whole experiment. Moreover, power Doppler allows the study of blood flow that reflects inflammatory activity within the synovium of arthritic joints. Our aim was to determine whether ultrasonography analysis could accurately detect arthritis lesions in a mouse model of rheumatoid arthritis, namely collagen-induced arthritis. METHODS: Collagen-induced arthritis was induced in 28 mice by immunising with collagen type II. Every week for 8 weeks, ultrasonography and Doppler analysis were performed on knees and ankles of all mice using the ultrasound biomicroscope (UBM), which is particularly dedicated to studying the mouse. Clinical and histological evaluations were performed as usual. RESULTS: We established a semiquantitative analysis by setting an UBM scoring. UBM grades were correlated to clinical and histological scores of arthritis. Vascularisation within the synovium could be estimated by power Doppler analysis and a semiquantitative vascularisation scale was established, which allowed us to show a good correlation between vascularisation scores and histological or clinical scores of arthritis. CONCLUSIONS: This is one of the first studies that shows it is possible to visualise a selected set of joints in a small animal using UBM analysis. It provides new perspectives in evaluating experimental models of rheumatoid arthritis and other joint diseases.

Bloquel C, Denys A, Boissier MC, Apparailly F, Bigey P, Scherman D, Bessis N. · Inserm, Eri-18, F-93017 Bobigny, France. · J Gene Med. · Pubmed #17912759 No free full text.

Abstract: BACKGROUND: Anti-inflammatory gene therapy is promising in inflammatory diseases such as rheumatoid arthritis (RA). We have previously demonstrated that intra-muscular (i.m.) electrotransfer (ET) of plasmids encoding three different human tumor necrosis factor-alpha-soluble receptor I variants (hTNFR-Is) exert protective effects in an experimental RA model. However, such a systemic approach could be responsible for side effects. The present study aimed at performing an intra-articular (i.a.) gene therapy by electrotransfer using the hTNFR-Is plasmids. METHODS AND RESULTS: We evaluated targeting of mice joints by CCD optical imaging after i.a. ET of a luciferase-encoding plasmid and we showed that ET led to strongly increased transgene expression in a plasmid dose-dependent manner. Moreover, articular and seric hTNFR-Is was detectable for 2 weeks. As expected, systemic hTNFR-Is rates were lower after i.a. ET than after i.m. ET. A longer protein secretion could be achieved with several i.a. ETs. Also, we observed that hTNFR-Is expression within arthritic joints was slightly higher than in normal joints. CONCLUSIONS: In collagen-induced arthritis (CIA), a mouse model for RA, we demonstrated that hTNFR-Is/mIgG1-encoding plasmid i.a. ET decreased joint destruction in the ankles. In conclusion, our results suggest that local TNFR-Is gene therapy may play a role in decreasing joint destruction in CIA.

Boissier MC, Lemeiter D, Clavel C, Valvason C, Laroche L, Begue T, Bessis N. · INSERM, Eri-18, F-93017 Bobigny, France. · Hum Gene Ther. · Pubmed #17532727 No free full text.

Abstract: Intraarticular gene transfer with adeno-associated viral (AAV) vectors may allow efficient therapeutic transgene expression within the joint in diseases such as rheumatoid arthritis (RA), allowing high expression of the protein within the joint, preventing both systemic diffusion and side effects. However, humans demonstrate antibodies against AAV, which can influence gene transfer. To better understand critical obstacles to intraarticular gene therapy with AAV, we have previously shown that synovial fluid (SF) contains IgG to AAV that neutralizes chondrocyte infection in vitro. Our objective was therefore to compare neutralization exerted by SF from RA patients for four different AAV serotypes (AAV serotypes 1, 2, 5, and 8) on human primary synoviocytes. Serotype 2 infected synoviocytes most efficiently followed, in decreasing order, by serotypes 1, 5, and 8. SF from all patients partially inhibited infection of synoviocytes by at least one of the four serotypes. Infection with serotypes 1 and 2 was the most inhibited by SF, whereas inhibition was weak for serotypes 5 and 8. Last, we have shown that inhibition of AAV1/interleukin (IL)-4 infection of synoviocytes by SF could be reversed by increasing the number of AAV1/IL-4 particles, with a dose-dependent effect. We conclude that the most infectious AAV serotypes (1 and 2) in synoviocytes are also the serotypes most neutralized by SF. Thus, serotype 5 seems to demonstrate the best infection efficiency:immunogenicity ratio for local use in articular diseases. These data may be useful for tailoring intraarticular AAV-mediated gene therapy to individual patients.

Bessis N, Lemeiter D, Laroche L, Fournier C, Huizinga T, Brok H, 't Hart B, Boissier MC. · Inserm, Eri-18, F-93017, Bobigny, France. · Joint Bone Spine. · Pubmed #17224293 No free full text.

Abstract: OBJECTIVES: Gene therapy using cells as vectors to achieve secretion of therapeutic proteins may hold promise in the treatment of chronic diseases. Cell-based gene therapy with xenogeneic cells secreting antiinflammatory cytokines (IL-4, IL-13, or IL-1 receptor type II) has been found effective in mice with collagen-induced arthritis (CIA), a model for human rheumatoid arthritis. Autologous cells engineered to produce antiinflammatory cytokines were also effective in the mouse CIA model. In all these experiments, the cells were grafted into the subcutaneous tissue of the back, resulting in systemic treatment. To evaluate the feasibility of cell-based gene therapy confined to the joints, we performed intraarticular injections of autologous cells in a rhesus monkey with CIA, a model more similar to human RA. METHODS: We prepared ex vivo cultures of skin fibroblasts from the animal then transfected the cells with a plasmid carrying the lacZ gene. We injected these marker cells into metacarpophalangeal, metatarsophalangeal, and interphalangeal joints. RESULTS: Kinetic evaluation of synovial tissue X-gal labeling, which reflected reported gene expression by skin fibroblasts present within the synovium, showed significant labeling by transfected cells up to 6 days after intraarticular injection. Xenogeneic fibroblasts (Chinese hamster ovary cells) injected intraarticularly were also detected within synovial specimens; however, labeling intensity was less marked than with autologous cells. Our findings establish the feasibility of skin fibroblast grafting into the synovium. CONCLUSION: This preliminary study opens the door to studies of heterotopic autologous transfected cells for the treatment of CIA in monkeys by direct gene transfer within joints.