apyrase and Arthritis--Rheumatoid

Rheumatoid arthritis (RA) is a classic inflammatory autoimmune disease. Local joint destruction and extra-articular manifestations of RA deeply compromise the life quality of the affected patients. RA immunopathogenesis depends on continuous immunogenic activation in which the purinergic system participates. The purinergic system comprises the signaling and metabolism of purines such as adenosine triphosphate (ATP) and adenosine. ATP signaling is involved in the activation and maintenance of the inflammatory state of RA through the activation of P2X7 and the production of cytokines, which orchestrate the pathogenesis of RA. The breakdown of ATP through the CD39/CD73 axis produces adenosine, which mostly inhibits the inflammatory process through activation of specific P1 receptors. Adenosine is hydrolyzed by adenosine deaminase (ADA) that interacts with other molecules playing additional roles in this disease. This review explores the release, metabolism, and the effects of binding of ATP and adenosine to their respective receptors in the context of RA, as well as their potential use as biomarkers and therapeutic targets.

Topics: 5'-Nucleotidase; Adenosine; Adenosine Triphosphate; Animals; Apyrase; Arthritis, Rheumatoid; Biomarkers; GPI-Linked Proteins; Humans; Receptors, Purinergic P2X7; Signal Transduction

Topics: Adenosine; Apyrase; Arthritis, Rheumatoid; B-Lymphocytes; B-Lymphocytes, Regulatory; Case-Control Studies; Cytokines; Disease Management; Disease Susceptibility; Gene Expression; Humans; Immunomodulation; Lymphocyte Activation; Lymphocyte Count; T-Lymphocytes; Treatment Outcome

Objective To investigate the levels and function status of CD4

Topics: 5'-Nucleotidase; Apyrase; Arthritis, Rheumatoid; Case-Control Studies; Forkhead Transcription Factors; GPI-Linked Proteins; Humans; Synovial Fluid; T-Lymphocytes, Regulatory; Transforming Growth Factor beta1

We sought to investigate whether genetic effects on response to TNF inhibitors (TNFi) in rheumatoid arthritis (RA) could be localised by considering known genetic susceptibility loci for relevant traits and to evaluate the usefulness of these genetic loci for stratifying drug response.. We studied the relation of TNFi response, quantified by change in swollen joint counts ( Δ SJC) and erythrocyte sedimentation rate ( Δ ESR) with locus-specific scores constructed from genome-wide assocation study summary statistics in 2938 genotyped individuals: 37 scores for RA; scores for 19 immune cell traits; scores for expression or methylation of 93 genes with previously reported associations between transcript level and drug response. Multivariate associations were evaluated in penalised regression models by cross-validation.. We detected a statistically significant association between Δ SJC and the RA score at the. The association with the CD39 trait is difficult to interpret because patients with RA are often prescribed TNFi after failing to respond to methotrexate. The CD39 and

Topics: Adult; Aged; Antigens, CD; Antirheumatic Agents; Apyrase; Arthritis, Rheumatoid; Biological Products; CD40 Antigens; Cohort Studies; Female; Gene Expression Regulation; Genome-Wide Association Study; Humans; Male; Middle Aged; Molecular Targeted Therapy; Multivariate Analysis; Predictive Value of Tests; Prognosis; Quantitative Trait Loci; Regression Analysis; Treatment Outcome; Tumor Necrosis Factor Inhibitors

Bone destruction is one of many severe complications that occurs in patients with rheumatoid arthritis (RA) and current therapies are unable to cure this manifestation. This study here aims to determine whether GMSC can directly inhibit osteoclast formation and eventually attenuate osteoclastogenesis and bone erosion in an inflammatory milieu.. GMSC were co-cultured with osteoclast precursors with or without CD39 inhibitor, CD73 inhibitor or adenosine receptors inhibitors pretreatment and osteoclast formation were evaluated in vitro. 2×10^6 GMSC per mouse were transferred to CIA mice and pathology scores, the frequency of osteoclasts, bone erosion in joints were assessed in vivo.. GMSC but not control cells, markedly suppressed human or mice osteoclastogenesis in vitro. GMSC treatment also resulted in a dramatically decreased level of NF-κB p65/p50 in osteoclasts in vitro. Infusion of GMSC to CIA significantly attenuated the severity of arthritis, pathology scores, frequency of osteoclasts, particularly bone erosion, as well as a decreased expression of RANKL in synovial tissues in vivo. Blockade of CD39/CD73 or adenosine receptors has significantly abrogated the suppressive ability of GMSC in vitro and therapeutic effect of GMSC on bone erosion during CIA in vivo.. GMSC inhibit osteoclast formation in vitro and in vivo partially via CD39-CD73-adenosine signals. Manipulation of GMSC may have a therapeutic implication on rheumatoid arthritis and other bone erosion related diseases. FUND: This study was supported by grants from the National Key R&D Program of China (2017YFA0105801 to F.H); the Zhujiang Innovative and Entrepreneurial Talent Team Award of Guangdong Province (2016 ZT 06S 252 to F·H) and National Institutes of Health (R01 AR059103, R61 AR073409 and NIH Star Award to S.G.Z).

Topics: Adenosine; Animals; Antigens, CD; Apyrase; Arthritis, Experimental; Arthritis, Rheumatoid; Autoimmune Diseases; Biomarkers; Cell Line; Female; Fibroblasts; Gingiva; Humans; Mesenchymal Stem Cells; Mice; Osteoclasts; Osteogenesis; Signal Transduction; Tomography, X-Ray Computed

Rheumatoid arthritis (RA) is an autoimmune arthropathy characterized by chronic articular inflammation. Methotrexate (MTX) remains the first-line therapy for RA and its anti-inflammatory effect is associated with the maintenance of high levels of extracellular adenosine (ADO). Nonetheless, up to 40% of RA patients are resistant to MTX treatment and this is linked to a reduction of CD39 expression, an ectoenzyme involved in the generation of extracellular ADO by ATP metabolism, on circulating regulatory T cells (Tregs). However, the mechanism mediating the reduction of CD39 expression on Tregs is unknown. Here we demonstrated that the impairment in TGF-β signalling lead to the reduction of CD39 expression on Tregs that accounts for MTX resistance. TGF-β increases CD39 expression on Tregs via the activation of TGFBRII/TGFBRI, SMAD2 and the transcription factor CREB, which is activated in a p38-dependent manner and induces CD39 expression by promoting ENTPD1 gene transcription. Importantly, unresponsive patients to MTX (UR-MTX) show reduced expression of TGFBR2 and CREB1 and decreased levels of p-SMAD2 and p-CREB in Tregs compared to MTX-responsive patients (R-MTX). Furthermore, RA patients carrying at least one mutant allele for rs1431131 (AT or AA) of the TGFBR2 gene are significantly (p = 0.0006) associated with UR-MTX. Therefore, we have uncovered a molecular mechanism for the reduced CD39 expression on Tregs, and revealed potential targets for therapeutic intervention for MTX resistance.

Topics: Adenosine Triphosphate; Adult; Aged; Antigens, CD; Antirheumatic Agents; Apyrase; Arthritis, Rheumatoid; Cells, Cultured; Cyclic AMP Response Element-Binding Protein; Drug Resistance; Female; Gene Expression Regulation; Gene Frequency; Humans; Male; Methotrexate; Middle Aged; Polymorphism, Single Nucleotide; Receptor, Transforming Growth Factor-beta Type I; Receptor, Transforming Growth Factor-beta Type II; Signal Transduction; Smad2 Protein; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

Nearly one-third of patients with rheumatoid arthritis (RA) do not respond to Methotrexate (MTX), the first-line therapy in RA. CD39, an ectonucleotidase highly expressed on regulatory T cells (Tregs), is responsible for production of adenosine, an important anti-inflammatory mediator of MTX action. Higher expression of CD39 on Tregs improves their suppressive capacity. Therefore, we aimed to study the role of CD39. Higher frequencies of CD39

Topics: Adult; Antirheumatic Agents; Apyrase; Arthritis, Rheumatoid; Biomarkers, Pharmacological; CD4 Lymphocyte Count; Drug Monitoring; Female; Flow Cytometry; Humans; Male; Methotrexate; Middle Aged; Pharmacogenomic Testing; Pharmacogenomic Variants; Polymorphism, Single Nucleotide; T-Lymphocytes, Regulatory; Treatment Outcome

This study aimed to identify CD4. Synovial fluids from 34 RA, 33 osteoarthritis patients and 42 healthy individuals were analyzed by flow cytometry. Human fibroblast-like synoviocytes and peripheral blood mononuclear cells were cultured with or without isolated MPs, chemokines and cytokines were measured by ELISA.. CD4

Topics: 5'-Nucleotidase; Adult; Antigens, CD; Apyrase; Arthritis, Rheumatoid; Biomarkers; Case-Control Studies; CD4-Positive T-Lymphocytes; Cell-Derived Microparticles; Chemokines; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Humans; Leukocytes, Mononuclear; Male; Middle Aged; NK Cell Lectin-Like Receptor Subfamily B; Osteoarthritis; Synovial Fluid

The effect of vitamin D3 in oral solution (VD3 ) and vitamin D3 -loaded nanocapsules (NC-VD3 ) was analysed in animals with complete Freund's adjuvant (CFA) induced arthritis (AR). For this purpose, we evaluated scores for arthritis, thermal hyperalgesia and paw oedema, as well as histological analyses and measurements of the activity of the ectonucleoside triphosphate diphosphohydrolase (E-NTPDase) and ecto-adenosine deaminase (E-ADA) enzymes in rat lymphocytes. Haematological and biochemical parameters were also determined. The doses administered were 120 UI/day of VD3 and 15.84 UI/day of NC-VD3 . Fifteen days after the induction of AR, the groups were treated for 15 days with vitamin D3 . The results demonstrated that VD3 was able to reduce arthritis scores, thermal hyperalgesia and paw oedema in rats with CFA-induced arthritis. However, treatment with NC-VD3 did not reduce arthritis scores. The histological analyses showed that both formulations were able to reduce the inflammatory changes induced by CFA. The activity of E-NTPDase in rat lymphocytes was higher in the AR compared with the control group, while the activity of E-ADA was lower. This effect was reversed after the 15-day treatment. Data from this study indicates that both forms of vitamin D3 seem to contribute to decreasing the inflammatory process induced by CFA, possibly altering the activities of ectoenzymes. Copyright © 2016 John Wiley & Sons, Ltd.. The effects promoted by both formulations of vitamin D3 , either in oral solution or nanoencapsulated form, strongly suggests the softening of the inflammatory process induced by complete Freund's adjuvant (CFA), possibly altering the E-NTPDase and E-ADA activities. However, it is known that vitamin D has a beneficial effect on the modulation of the immune system components responsible for the inflammatory process. Moreover, the establishment of responses to treatment with vitamin D3 may provide an alternative for inhibiting the proinflammatory response, assisting in our understanding of the immunopathology of this disease and possibly improving the signs and symptoms that hinder the quality of life of patients with rheumatoid arthritis.. Evaluation of the effects on the E-NTPDase and E-ADA activities in an animal model of induced arthritis. Two formulations of vitamin D3 were used: form oral solution and nanoencapsulated. Vitamin D3 seems to contribute to the inflammatory process induced by CFA. Vitamin D3 possibly alters the E-NTPDase and E-ADA activities. Vitamin D3 may be an alternative supplementary treatment for chronic arthritis.

Topics: Adenosine Deaminase; Administration, Oral; Animals; Anti-Inflammatory Agents; Antigens, CD; Apyrase; Arthritis, Rheumatoid; Cholecalciferol; Disease Models, Animal; Female; Freund's Adjuvant; Lymphocytes; Nanocapsules; Nanoparticles; Rats, Wistar; Solutions

IL-33 is strongly involved in several inflammatory and autoimmune disorders with both pro- and anti-inflammatory properties. However, its contribution to chronic autoimmune inflammation, such as rheumatoid arthritis, is ill defined and probably requires tight regulation. In this study, we aimed at deciphering the complex role of IL-33 in a model of rheumatoid arthritis, namely, collagen-induced arthritis (CIA). We report that repeated injections of IL-33 during induction (early) and during development (late) of CIA strongly suppressed clinical and histological signs of arthritis. In contrast, a late IL-33 injection had no effect. The cellular mechanism involved in protection was related to an enhanced type 2 immune response, including the expansion of eosinophils, Th2 cells, and type 2 innate lymphoid cells, associated with an increase in type 2 cytokine levels in the serum of IL-33-treated mice. Moreover, our work strongly highlights the interplay between IL-33 and regulatory T cells (Tregs), demonstrated by the dramatic in vivo increase in Treg frequencies after IL-33 treatment of CIA. More importantly, Tregs from IL-33-treated mice displayed enhanced capacities to suppress IFN-γ production by effector T cells, suggesting that IL-33 not only favors Treg proliferation but also enhances their immunosuppressive properties. In concordance with these observations, we found that IL-33 induced the emergence of a CD39(high) Treg population in a ST2L-dependent manner. Our findings reveal a powerful anti-inflammatory mechanism by which IL-33 administration inhibits arthritis development.

Topics: Animals; Antigens, CD; Apyrase; Arthritis, Experimental; Arthritis, Rheumatoid; Autoimmune Diseases; Collagen; Cytokines; Disease Models, Animal; Eosinophils; Interleukin-33; Mice; Mice, Inbred DBA; Spleen; T-Lymphocytes, Regulatory; Th2 Cells

Rheumatoid arthritis (RA) is an inflammatory autoimmune disease characterized by joint destruction and severe morbidity. Methotrexate (MTX) is the standard first-line therapy of RA. However, about 40% of RA patients are unresponsive to MTX treatment. Regulatory T cells (Tregs, CD4(+)CD25(+)FoxP3(+)) are thought to play an important role in attenuating RA. To investigate the role of Tregs in MTX resistance, we recruited 122 RA patients (53 responsive, R-MTX; 69 unresponsive, UR-MTX) and 33 healthy controls. Three months after MTX treatment, R-MTX but not UR-MTX showed higher frequency of peripheral blood CD39(+)CD4(+)CD25(+)FoxP3(+) Tregs than the healthy controls. Tregs produce adenosine (ADO) through ATP degradation by sequential actions of two cell surface ectonucleotidases: CD39 and CD73. Tregs from UR-MTX expressed a lower density of CD39, produced less ADO, and had reduced suppressive activity than Tregs from R-MTX. In a prospective study, before MTX treatment, UR-MTX expressed a lower density of CD39 on Tregs than those of R-MTX or control (P < 0.01). In a murine model of arthritis, CD39 blockade reversed the antiarthritic effects of MTX treatment. Our results demonstrate that MTX unresponsiveness in RA is associated with low expression of CD39 on Tregs and the decreased suppressive activity of these cells through reduced ADO production. Our findings thus provide hitherto unrecognized mechanism of immune regulation in RA and on mode of action of MTX. Furthermore, our data suggest that low expression of CD39 on Tregs could be a noninvasive biomarker for identifying MTX-resistant RA patients.

Topics: 5'-Nucleotidase; Adenosine; Animals; Antigens, CD; Apyrase; Arthritis, Experimental; Arthritis, Rheumatoid; Biomarkers; Drug Resistance; Humans; Lymphocyte Count; Methotrexate; Mice, Inbred C57BL; T-Lymphocytes, Regulatory; Th1 Cells; Th17 Cells

In autoimmune diseases such as rheumatoid arthritis (RA), regulatory T cells (Tregs) fail to constrain autoimmune inflammation; however, the reasons for this are unclear. We investigated T cell regulation in the RA joint. Tregs from RA synovial fluid suppressed autologous responder T cells; however, when compared with Tregs from healthy control peripheral blood, they were significantly less suppressive. Despite their reduced suppressive activity, Tregs in the RA joint were highly proliferative and expressed FOXP3, CD39, and CTLA-4, which are markers of functional Tregs. This suggested that the reduced suppression is due to resistance of RA synovial fluid responder T cells to Treg inhibition. CD161(+) Th17 lineage cells were significantly enriched in the RA joint; we therefore investigated their relative susceptibility to Treg-mediated suppression. Peripheral blood CD161(+) Th cells from healthy controls were significantly more resistant to Treg-mediated suppression, when compared with CD161(-) Th cells, and this was mediated through a STAT3-dependant mechanism. Furthermore, depletion of CD161(+) Th cells from the responder T cell population in RA synovial fluid restored Treg-mediated suppression. In addition, CD161(+) Th cells exhibited pathogenic features, including polyfunctional proinflammatory cytokine production, an ability to activate synovial fibroblasts, and to survive and persist in the inflamed and hypoxic joint. Because CD161(+) Th cells are known to be enriched at sites of autoinflammation, our finding that they are highly proinflammatory and resistant to Treg-mediated suppression suggests an important pathogenic role in RA and other autoimmune diseases.

Topics: Adult; Aged; Aged, 80 and over; Antigens, CD; Apyrase; Arthritis, Rheumatoid; Autoimmunity; Cell Lineage; CTLA-4 Antigen; Cytokines; Female; Forkhead Transcription Factors; Gene Expression Regulation; Humans; Joints; Lymphocyte Depletion; Male; Middle Aged; NK Cell Lectin-Like Receptor Subfamily B; Primary Cell Culture; Signal Transduction; STAT3 Transcription Factor; Synovial Fluid; T-Lymphocytes, Regulatory; Th17 Cells

The rationale for blocking interleukin-6 (IL-6) in rheumatoid arthritis (RA) lies chiefly in the proinflammatory effect of this cytokine. Few studies have evaluated the consequences of anti-IL-6 receptor (IL-6R) antibody treatment on Treg cells. This study was undertaken to elucidate the mechanism of action of anti-IL-6R antibody treatment by studying the effects on Treg cells in an experimental arthritis model and in patients with RA.. Mice with collagen-induced arthritis (CIA) were treated with a mouse anti-IL-6R antibody (MR16-1), and changes in Treg, Th1, and Th17 cells were assessed at key time points during the course of the disease. Peripheral blood from 15 RA patients was collected on day 0 and after 3 months of tocilizumab treatment for flow cytometry analysis of Th17 and Treg cells.. In MR16-1-treated mice, Th17 cell frequencies were unchanged, whereas Treg cell frequencies were increased. The Treg cell phenotype showed marked changes, with an increase in the frequency of CD39+ Treg cells in the lymph nodes and spleen. Interestingly, similar CD39+ Treg cell expansion was observed in RA patients who were tocilizumab responders at 3 months, with no change in Th17 cell frequency. Moreover, fluorescence-activated cell-sorted CD39+ Treg cells from responder RA patients were functionally able to suppress the proliferation of conventional T cells.. In both CIA and RA, the frequency of functionally suppressive CD39+ Treg cells is increased as a result of anti-IL-6R treatment, whereas Th17 cells are unaffected. The modification of Treg cell frequency and phenotype may be one of the mechanisms involved in the therapeutic effect of IL-6 blockade in RA.

Topics: Animals; Antibodies, Anti-Idiotypic; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Antigens, CD; Apyrase; Arthritis, Experimental; Arthritis, Rheumatoid; Cell Proliferation; Disease Models, Animal; Female; Humans; Male; Mice; Mice, Inbred DBA; Middle Aged; Phenotype; Receptors, Interleukin-6; T-Lymphocytes, Regulatory; Th17 Cells

Topics: Animals; Antigens, CD; Apyrase; Arthritis, Experimental; Arthritis, Rheumatoid; Female; Humans; Male; Receptors, Interleukin-6; T-Lymphocytes, Regulatory

Topics: Animals; Antigens, CD; Apyrase; Arthritis, Experimental; Arthritis, Rheumatoid; Female; Humans; Male; Receptors, Interleukin-6; T-Lymphocytes, Regulatory

Topics: Animals; Antigens, CD; Apyrase; Arthritis, Experimental; Arthritis, Rheumatoid; Female; Humans; Male; Receptors, Interleukin-6; T-Lymphocytes, Regulatory

Treg cells are important for the maintenance of self-tolerance and are implicated in autoimmunity. Despite enrichment of Treg cells in joints of rheumatoid arthritis (RA) patients, local inflammation persists. As expression of the ATP-hydrolyzing enzymes CD39 and CD73 and the resulting anti-inflammatory adenosine production have been implicated as an important mechanism of suppression, we characterized FOXP3(+) Treg cells in blood and synovial fluid samples of RA patients in the context of CD39 and CD73 expression. Synovial FOXP3(+) Treg cells displayed high expression levels of rate-limiting CD39, whereas CD73 was diminished. FOXP3(+) CD39(+) Treg cells were also abundant in synovial tissue. Furthermore, FOXP3(+) CD39(+) Treg cells did not secrete the proinflammatory cytokines IFN-γ and TNF after in vitro stimulation in contrast to FOXP3(+) CD39(-) T cells. FOXP3(+) CD39(+) Treg cells could be isolated by CD39 and CD25 coexpression, displayed a demethylated Treg-specific demethylated region and coculture assays confirmed that CD25(+) CD39(+) T cells have suppressive capacity, while their CD39(-) counterparts do not. Overall, our data show that FOXP3(+) CD39(+) Treg cells are enriched at the site of inflammation, do not produce proinflammatory cytokines, and are good suppressors of many effector T-cell functions including production of IFN-γ, TNF, and IL-17F but do not limit IL-17A secretion.

Topics: Adult; Aged; Aged, 80 and over; Antigens, CD; Apyrase; Arthritis, Rheumatoid; Cell Separation; Female; Flow Cytometry; Humans; Interleukin-17; Male; Microscopy, Confocal; Middle Aged; Synovial Fluid; T-Lymphocyte Subsets

The purpose of this study was to investigate the activities of ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase; EC 3.6.1.5; CD39) and adenosine deaminase (E-ADA; EC 3.5.4.4) in lymphocytes from patients with rheumatoid arthritis (RA). Thirty patients diagnosed with RA through American College of Rheumatology criteria as well as 30 healthy patients were selected. Peripheral blood lymphocytes were isolated, and E-NTPDase and E-ADA activities were assayed. The results demonstrated an increased E-NTPDase activity (both ATP and ADP as substrates) and a decreased E-ADA activity in RA patients. These data suggest an organic effort to preserve the adenosine level, which is known to have anti-inflammatory and analgesic properties, working as a potent suppressor of immune response.

Topics: Adenosine Deaminase; Adenosine Diphosphate; Adenosine Triphosphate; Adult; Antigens, CD; Apyrase; Arthritis, Rheumatoid; Case-Control Studies; Cells, Cultured; Enzyme Assays; Female; Humans; Lymphocytes; Male; Middle Aged