Francesca Oliviero

Co-stimulatory modulation in rheumatoid arthritis: The role of (CTLA4-Ig) abatacept

Associations between rheumatoid arthritis (RA) susceptibility and polymorphism in multiple immunoregulatory genes suggest a role of altered T cell function in the disease. The growing relevance of the oxidative stress in RA synovitis, which results in a number of T cell signalling abnormalities, is reinforced by the demonstration of a direct NO inducing activity through the shared epitope of the HLA class II molecules HLA-DRbeta1, with secondary lymphocytes oxidative damage. Direct T cell/macrophage contact-dependent activation, one of the driving mechanisms of synovitis, is mediated by co-stimulatory molecules as well as cell membrane cytokines and may also result in an impaired suppressive function of T regulatory cells (Treg) in RA joints. The fusion of CTLA4 extracellular binding domain to the Fcgamma1 allows to obtain a soluble CTLA4 receptor, the dimeric recombinant human fusion protein abatacept (CTLA4-Ig). The improved knowledge of the CTLA4-B7 co-stimulation regulatory mechanisms by signals delivered into DCs and Tregs provides multiple potential targets for the abatacept treatment. CTLA4-Ig shows the capacity, either ex vivo or in vivo, to interrupt at multiple steps the ongoing inflammatory and destructive process, and to concur in restoring the immunoregulatory balance in RA.

Synovial effusion and synovial fluid biomarkers in psoriatic arthritis to assess intraarticular tumor necrosis factor-α blockade in the knee joint.

IntroductionThe purpose of this study was theevaluation of synovial effusion (SE), synovial fluid (SF) and synovial tissue (ST) biomarkers in relation to disease activity indexes to assess the response to intraarticular (IA) tumor necrosis factor (TNF)-α blockers in psoriatic arthritis (PsA).MethodsSystemic and local disease activity indexes (disease activity score (DAS); the Ritchie articular index (mRAI), erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP); Thompson articular (THOMP) and joint articular (KJAI)-Index ) and ST samples were assessed at baseline, throughout treatment, and during the follow-up in 14 patients affected with PsA who underwent IA injections (0.5 ml to 12.5 mg) in the knee joint of etanercept (E) or placebo (P) once every two weeks for a 10-week period. Total SF white blood cell (WBC) counts (WBC/μl) and SF cytokine/chemokine (CK/CCK) levels were measured before IA-E at baseline, after IA-E, and as long as there were adequate amounts of SF for knee aspiration (post). Characterization of synovial mononuclear cell infiltration and synovial vessels was carried out in 8 out of 14 knees by staining serial sections of synovial tissue biopsies for CD45, CD3, CD68, CD31 and CD105.ResultsAt baseline, CRP and/or ESR were significantly correlated with SF-CK (interleukin- (IL-)1β, IL-1Ra, IL-6, IL-8) and CCK (CCL3). Post-IA injections, there was a decrease in SE in the knees in which aspiration following IA-E injection was possible as well as a significant reduction in SF WBC/μl and in SF-CK (IL-1β, IL-1Ra, IL-6 and IL-22). Pre- and post-IA-E injections, there were significant correlations between ST markers and SF-CK (IL-1β with CD45; IL-1β and IL-6 with CD31) and between SF-CCK (CCL4 and CCL3 with CD3). At the end of the study, there was a significant reduction in disease activity indexes (CRP, DAS, RAI, THOMP, KJAI) as well as in the ST markers (CD45; CD3).ConclusionsSynovial effusion regression is a reliable indicator of the response to IA TNF-α blockers in PsA patients as it is confirmed by the correlation between SF biomarkers to disease activity and synovial tissue inflammation.

Cytokine levels in human synovial fluid during the different stages of acute gout: role of transforming growth factor β1 in the resolution phase.

Objectives To determine the most relevant parameters in synovial fluid (SF) during the various stages of acute gout. Methods SFs from 38 gouty patients were analysed for white blood cell (WBC) count, percentage of polymorphonuclear cells (PMNs) and levels of interleukin 1β (IL-1β), IL-6, IL-8, tumour necrosis factorα (TNFα) and transforming growth factor β1 (TGFβ1). Patients were divided into three groups according to the length of time since onset of the attack: phase I (0–48 h), phase II (days 3–4) and phase III (days 5–7). Results Levels of WBCs were similar in SFs from phases I and II, while phase III showed the lowest WBC count. Percentages of PMNs were raised in all SFs. None of the cytokines analysed differed between phases I and II except for TGFβ1, which was higher in phase II. IL-1β, IL-6 and TNFα were higher in group 1 than in group 3. Levels of all the cytokines assessed, with the exception of TGFβ1, were significantly lower in phase III than in phase II IL-1β, p<0.05; IL-6, p<0.01; IL-8, p<0.001; TNFα, p<0.05).TGFβ1 levels were highest in SFs from phase III. Conclusion Cytokine levels in SFs may change depending on the different stages of acute gout, highlighting the role of TGFβ1 in the resolution of gout.

Gout as autoinflammatory disease: New mechanisms for more appropriated treatment targets

Gout is probably one of the oldest diseases affecting men. This is not surprising especially for the active role that innate immunity seems to play in its pathogenesis. It is fascinating to observe as this ancestral mechanism of defense feels that microcrystals are a danger, quite similar to infectious agents. New advances have revealed that at the base of the powerful inflammatory reaction stimulated by monosodium urate crystals there are many complexes cellular mechanisms, mainly involving inflammasome and toll-like receptors. Subsequently, there is an early increase of proinflammatory cytokines responsible for the acute attack, followed in few days by their reduction along with an increase of anti-inflammatory cytokines, probably main actors of the resolution phase. New targets have also been identified for the reduction of hyperuricemia, the prerequisite of gout, in order to prevent new attacks and the deposition of urate crystals in and around the joints. All these aspects, leading to deeper insight, have suggested new treatments, some of which are already available while others are likely to become available in the near future.

New Biochemical Insights into the Pathogenesis of Osteoarthritis and the Role of Laboratory Investigations in Clinical Assessment

Osteoarthritis (OA) is among the most frequent diseases in the population and a common cause of pain and disability in adults. The principal disease hallmarks for assessment of OA are still clinical observation and radiographic aspects. However, the efficacy of therapeutic interventions is complicated by the time required to observe radiographic signs, useful for both diagnosis and assessment. Thus, laboratory markers have received growing attention in recent years, in an attempt to improve diagnosis, assessment of disease activity and severity, and evaluation of therapeutic effects. Many biomarkers have been proposed, in particular those reflecting cartilage and bone turnover and synovitis. Among these, COMP, antigenic keratan sulphate, hyaluronan, YKL-40, type III collagen N-propeptide, and urinary glucosyl-galactosyl pyridinoline appear to be the most promising. However, serum or urinary determinations of these molecules are difficult to interpret adequately due to their complex metabolism. New ultrasensitive methods for C-reactive protein have improved the usefulness of this marker, especially in the assessment of disease activity. Routine examination of synovial fluid is still essential for diagnosis and includes leukocyte count and crystal detection; specialized testing includes the evaluation of the levels of markers of local inflammation such as metalloproteinases and cytokines, which appear to be crucial to the pathogenesis of OA.

High-density lipoproteins downregulate CCL2 production in human fibroblast-like synoviocytes stimulated by urate crystals

IntroductionTo investigate whether monosodium urate (MSU) crystals induce the production of CCL2 (monocyte chemoattractant protein-1; MCP-1) in human fibroblast-like synoviocytes (FLS) and whether this mechanism would be affected by high-density lipoproteins (HDL).MethodsHuman FLS isolated from synovial tissue explants were stimulated with MSU crystals (0.01 to 0.5 mg/ml) or interleukin (IL)-1β (10 pg/ml) in the presence or absence of HDL (50 and 100 μg/ml). The production and expression of CCL2 was evaluated with ELISA, confocal microscopy, immunofluorescence microscopy, chemotaxis assay, and real-time quantitative PCR.ResultsExposure of FLS to MSU crystals induced CCL2 accumulation in culture medium in a dose- and time-dependent manner, reaching a plateau at 50 to 75 μg/ml MSU crystals and 20 to 24 hours. Although low, the induced CCL2 levels were sufficient to trigger mononuclear cell migration. In resting FLS, CCL2 was localized in small cytoplasmic vesicles whose number diminished with MSU crystal stimulation. Concomitantly, MSU crystals triggered the induction of CCL2 mRNA expression. All these processes were inhibited by HDL, which cause a 50% decrease in CCL2 mRNA levels and a dose-dependent inhibition of the release of CCL2. Similar results were obtained when FLS were pretreated with HDL and washed before activation by MSU crystals or IL-1β, suggesting a direct effect of HDL on the FLS activation state.ConclusionsThe present results demonstrate that MSU crystals induce FLS to release CCL2 that is stored in vesicles in resting conditions. This mechanism is inhibited by HDL, which may limit the inflammatory process by diminishing CCL2 production and, in turn, monocytes/macrophages recruitment in joints. This study confirms the antiinflammatory functions of HDL, which might play a part in the limitation of acute gout attack.

New horizons in osteoarthritis

Osteoarthritis (OA), also known as degenerative joint disease, is the most frequent chronic musculoskeletal disease and the leading cause of disability in elderly persons. There are currently at least 27 million persons afflicted with OA in the United States, and the annual cost to society in medical care and wage loss is expected to reach nearly

JAK/STAT/PKCδ molecular pathways in synovial fluid T lymphocytes reflect the in vivo T helper-17 expansion in psoriatic arthritis

100 billion dollars by 2020, with consequent increased spending on its diagnosis and treatment, side effect prevention, and loss of productivity. Despite this enormous burden, many aspects of OA are still unknown, with implications not only in terms of diagnosis and assessment but also with regard to therapy. Awareness of this state of affairs has attracted many researchers to this field, making OA one of the most actively studied sectors of rheumatology. Although some clinicians are unaware of recent advances, there is a large body of publications indicating that much has been achieved. Major progress has been made in formulating better definitions of risk factors, in particular in indicating the responsibility of biomechanical and genetic factors, and, with regard to pathogenesis, underlining the role of subchondral bone, cytokines and proteinases. Assessment of OA activity and its progression has been improved with the advent of biomarkers and new imaging procedures, in particular sonography and magnetic resonance imaging (MRI), but also of better clinical instruments, including more reliable patient questionnaires. Information from ongoing studies may improve the to some extent incomplete definition of OA phenotypes. Finally, promising new horizons have been opened up even with regard to the treatment of OA, which is still for the most part unsatisfactory except for surgical replacement therapy. Numerous new substances have been formulated and the findings of trials studying their effects are encouraging, although much has yet to be done.

A comparative study of serum and synovial fluid lipoprotein levels in patients with various arthritides.

AbstractLooking to the sustained psoriatic arthritis (PsA) joint as a model of local human inflammation, this study was designed to assess the T lymphocyte signal transduction pathways potentially involved in this chronic immune-mediated inflammatory process, as characterized by direct ex vivo analysis of T helper (Th)-17 T effector (Teff) cell phenotypes in synovial fluid (SF) and peripheral blood (PB) of clinically active PsA patients. The reverse-phase protein arrays (RPPA) technique was employed to identify STAT3, STAT1, JAK1, JAK2, PKCδ and ERK1/2 phosphoprotein levels on total T cell lysates in SF samples of PsA patients. Frequencies of T CD4+IL-17A-F+ and T CD4+IL-23R+ Th17 cells were quantified in SF and matched PB of PsA patients by flow cytometry and compared with PB of healthy controls (HC). Increased levels of JAK1, STAT3, STAT1 and PKCδ phosphoproteins were found in SF T cells of PsA patients, compared with PB of HC. The expansion of T CD4+IL-17A-F+ cells, as well as of T CD4+ cells expressing IL-23Rp19 (T CD4+ IL-23R+), considered as the pathogenic phenotype of effector Th17 cells, was found to be confined to the joints of PsA patients, as the frequencies of both populations were significantly higher in SF than in matched PB, or in PB of HC. In conclusion, T lymphocyte signal transduction pathway mapping revealed an enhanced activation of JAK1/STAT3/STAT1 and PKCδ phosphoproteins that may drive the local inflammatory process, characterized by the in vivo expansion of T CD4+IL-17A-F+ and T CD4+IL-23R+ Th17 Teff cells in SF of clinically active joints of PsA patients.

Epithelial CXCR3-B regulates chemokines bioavailability in normal, but not in Sjögren's syndrome, salivary glands

UNLABELLED The aim of this study was to investigate apolipoprotein (apo) A-I, apo B, lipoprotein (Lp) (a), HDL-cholesterol (C), LDL-C, triglycerides (TG) and total cholesterol (TC) values in the serum and synovial fluid (SF) of untreated rheumatoid arthritis (RA), psoriatic arthritis (PsA), and osteoarthritis (OA) patients. METHODS Paired SF and serum samples were collected simultaneously from 14 patients with RA, 14 with PsA, and 16 with OA and tested for apo A-I, apo B, HDL-C, LDL-C, Lp(a), TC and TG. Serum C reactive protein (CRP) and amyloid A (SAA) levels were also determined. RESULTS The inflammatory arthritis patients had higher SF lipid levels with the exception of HDL. Reflecting increased synovial permeability, the lipid SF/serum ratio was always higher in RA and PsA with respect to OA patients. The positive correlation between serum and SF apo A-I, apo B, HDL-C, TG, and Lp(a) levels confirmed that there is lipoprotein diffusion into the SF. RA and PsA patients had lower concentrations of all serum lipids except for Lp(a) with respect to OA patients. The levels in the RA patients were similar to those in healthy matched controls, while the PsA patients had significantly lower apo A-I and HDL levels and higher apo B and LDL values. CONCLUSIONS Lipid diffusion into the joint cavity, which largely depends on the degree of inflammation, may contribute to modulating local inflammatory processes.