FVB/N (H2 q ) mouse is resistant to arthritis induction and exhibits a genomic deletion of T-cell receptor V beta gene segments

 Animal models of autoimmune diseases have been instrumental in advancing our understanding of autoimmunity in humans. Collagen-induced arthritis (CIA) in mice is an autoimmune disease model of rheumatoid arthritis. Susceptibility to CIA in mice is linked to genes of the major histocompatibility complex (MHC). CD4⁺ T cells that express the T-cell receptor (TCR) Tcra-V11.1 and/or Tcrb-V8.2 play a key role in the pathogenesis of arthritis in the DBA/1 mouse (H2 ^q ). We identified an inbred mouse strain, FVB/NJ (H2 ^q ), that is resistant to arthritis induction and exhibits a genomic deletion of certain Tcrb-V gene segments. We report a novel polymerase chain reaction-based method for the rapid identification of new mouse strains that exhibit germline Tcrb-V gene deletions. We mapped for the first time both the 5′ and 3′ breakpoints of the Tcrb-V deletion in the FVB/NJ, SWR, SJL, C57L, and C57BR strains to within 1.1 kilobases. Since there is an association between a particular Tcra-V allele (Tcra-V11.1 ^d ) and arthritis susceptibility in H2 ^q mouse strains, we examined the allelic polymorphisms of the Tcra-V11 gene subfamily members between the arthritis-susceptible DBA/1 mouse and the arthritis-resistant FVB/NJ mouse strain. The amino acid sequences of the Tcra-V11.1 alleles differ at two positions (codons 18 and 68). Therefore, the resistance of FVB/NJ mouse to arthritis induction may be due in part to Tcra-V11.1 coding sequence polymorphism and Tcrb-V8.2 gene segment deletion, as we have recently demonstrated in the case of SWR mouse strain. Received: 12 January 1999 / Revised: 17 March 1999     

Potential roles of hypochlorous acid and N-chloroamines in collagen breakdown by phagocytic cells in synovitis

We have tested the effects of the neutrophil/macrophage products, hypochlorous acid (HOCl) and N-chloroamines, on the structural integrity and proteolytic susceptibility of collagen to determine if these agents could play a role in inflammatory joint destruction. Rates of HOCl reaction with collagen, and collagen gelation were monitored by spectrophotometric methods. Direct fragmentation, and degradation by collagenase were measured by the release of acid-soluble counts from ³H-collagen. Physiologically relevant concentrations of HOCl (5–50 μM) reacted rapidly and quantitatively at several sites in the collagen polypeptide chain, causing extensive protein fragmentation and preventing collagen gelation. In contrast, reaction with (5–50 μM) N-chloroalanine induced little direct collagen fragmentation. Oxidative damage by N-chloroamines was, however, evident because collagen displayed greatly increased proteolytic susceptibility following N-chloroamine treatment. Collagen degradation by collagenase increased as much as 3-fold after exposure to N-chloroamine treatment. Collagen degradation by collagenase increased as much as 3-fold after exposure to N-chloroalanine. N-chloroleucine caused a small increase in proteolytic susceptibility, but N-chlorotaurine had no effect. Collagen fragmentation by HOCl, inhibition of gelation by HOCl, and N-chloroalanine-induced proteolytic susceptibility, all increased with linear kinetics at oxidant concentrations of 5 μM to 1.0 mM. In synovitis, phagocytes expose collagen to HOCl, N-chloroamines, and collagenase. It is known that HOCl can activate neutrophil procollagenase. Based on our new findings, we propose a model of inflammatory joint destruction that also includes collagen fragmentation, and increased susceptibility of collagen to degradation by collagenase. It may also be possible that taurine exerts a protective effect against HOCL/OCL⁻ damage by reacting to form what appears to be essentially an inert N-chloroamine. The validity of this model must now be tested.     

Foot structure and knee joint kinetics during walking with and without wedged footwear insoles

The relationship between static foot structure characteristics and knee joint biomechanics during walking, or the biomechanical response to wedged insoles are currently unknown. In this study, 3D foot scanning, dual X-ray absorptiometry and gait analysis methods were used to determine structural parameters of the foot and assess their relation to knee joint loading and biomechanical response to wedged insoles in 30 patients with knee osteoarthritis. In multiple linear regression models, foot fat content, height of the medial longitudinal arch and static hind foot angle were not associated with the magnitude of the knee adduction moment (R² = 0.24, p = 0.060), knee adduction angular impulse (R² = 0.21, p = 0.099) or 3D resultant knee moment (R² = 0.23, p = 0.073) during gait. Furthermore, these foot structure parameters were not associated with the patients’ biomechanical response to medial or lateral wedge footwear insoles (all p < 0.01). These findings suggest that static foot structure is not associated with gait mechanics at the knee, and that static foot structure alone cannot be utilized to predict an individual’s biomechanical response to wedged footwear insoles in patients with knee osteoarthritis.     

B cells from Patients with Rheumatoid Arthritis Show Conserved CD39-Mediated Regulatory Function and increased CD39 Expression After Positive Response to Therapy

Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by progressive joint destruction associated with increased pro-inflammatory mediators. In inflammatory microenvironments, exogenous ATP (eATP) is hydrolyzed to adenosine, which exerts immunosuppressive effects, by the consecutive action of the ectonucleotidases CD39 and CD73. Mature B cells constitutively express both ectonucleotidases, converting these cells to potential suppressors. Here, we assessed CD39 and CD73 expression on B cells from treated or untreated patients with RA. Neither the frequency of CD73⁺CD39⁺ and CD73^-CD39⁺ B cell subsets nor the levels of CD73 and CD39 expression on B cells from untreated or treated RA patients showed significant changes in comparison to healthy controls (HC). CpG+IL-2-stimulated B cells from HC or untreated RA patients increased their CD39 expression, and suppressed CD4⁺ and CD8⁺ T cell proliferation and intracellular TNF-production. A CD39 inhibitor significantly restored proliferation and TNF-producing capacity in CD4⁺ T cells, but not in CD8⁺ T cells, from HC and untreated RA patients, indicating that B cells from untreated RA patients conserved CD39-mediated regulatory function. Good responder patients to therapy (R-RA) exhibited an increased CD39 but not CD73 expression on B cells after treatment, while most of the non-responder (NR) patients showed a reduction in ectoenzyme expression. The positive changes of CD39 expression on B cells exhibited a negative correlation with disease activity and rheumatoid factor levels. Our results suggest modulating the ectoenzymes/ADO pathway as a potential therapy target for improving the course of RA.     

Targeting the Fas/FasL system in Rheumatoid Arthritis therapy: Promising or risky?

Rheumatoid Arthritis (RA) is a chronic inflammatory disease affecting synovial joints. Tumor necrosis factor (TNF) α is a key component of RA pathogenesis and blocking this cytokine is the most common strategy to treat the disease. Though TNFα blockers are very efficient, one third of the RA patients are unresponsive or present side effects. Therefore, the development of novel therapeutic approaches is required. RA pathogenesis is characterized by the hyperplasia of the synovium, closely associated to the pseudo-tumoral expansion of fibroblast-like synoviocytes (FLS), which invade and destroy the joint structure. Hence, depletion of RA FLS has been proposed as an alternative therapeutic strategy. The TNF family member Fas ligand (FasL) was reported to trigger apoptosis in FLS of arthritic joints by binding to its receptor Fas and therefore suggested as a promising candidate for targeting the hyperplastic synovial tissue. However, this cytokine is pleiotropic and recent data from the literature indicate that Fas activation might have a disease-promoting role in RA by promoting cell proliferation. Therefore, a FasL-based therapy for RA requires careful evaluation before being applied. In this review we aim to overview what is known about the apoptotic and non-apoptotic effects of Fas/FasL system and discuss its relevance in RA.     

Purine Metabolism and Clinical Status of Patients with Rheumatoid Arthritis Treated with Dipyridamole

The anti-inflammatory activities of methotrexate and sulphasalazine may be mediated by increases in endogenous adenosine levels. Since the vascular protective drug dipyridamole inhibits the uptake and metabolism of adenosine we have now tested this compound in patients with rheumatoid arthritis to assess its effects on their symptoms. Forty patients (aged 18–75 years) received dipyridamole 400 mg/day or placebo. The levels of adenosine and its major metabolites were determined by high performance liquid chromatography (HPLC) in blood samples taken at baseline and at monthly intervals during treatment for 6 months. After three months of treatment there was a significant reduction in the modified Health Assessment Questionnaire (mHAQ) score, but these effects were not maintained, and dipyridamole did not modify disease severity scores or the levels of adenosine and its metabolites. We conclude that the symptoms of rheumatoid arthritis were not modified by treatment with dipyridamole.