Cyclic adenosine 5′-monophosphate in synovial fluid of rheumatoid arthritis and osteoarthritis patients

The relationship between synovial fluid (SF) cAMP level and IL-18 and PGE2 SF levels in rheumatoid arthritis (RA) and osteoarthritis (OA) patients, and between SF cAMP level and disease as well as inflammatory activity in RA were investigated in 17 RA and 19 OA patients. Erythrocyte sedimentation rate (ESR), serum (S) C-reactive protein (CRP) level and SF IL-18 level were higher in RA than in OA patients. SF PGE2 level was similar in both groups. SF cAMP level was higher in OA than in RA patients. In RA patients, SF cAMP level showed negative correlation with Disease Activity Score including a 28-joint count and S CRP, ESR and SF IL-18 level. The results suggest that cAMP promotes anti-inflammatory response in RA and OA patients, which is higher in the latter. Promotion of anti-inflammatory response by cAMP elevating agents might be useful in the treatment of RA.     

Plasma and synovial fluid levels of granulocytal elastase-α-1-protease inhibitor complex in patients with rheumatoid arthritis

Summary Plasma and synovial fluid levels and granulocytal elastase--1-protease inhibitor complex (EIC) in patients with rheumatoid arthritis (RA) were measured by enzyme-linked immunoassay and the results compared with those in patients with osteoarthrosis (OA). It was found that the plasma and synovial fluid levels of EIC in RA patients were higher than those in OA patients. There was a positive correlation between plasma EIC level in RA patients in Lansbury's index score of disease activity, as this tends to be higher when titer of RAHA in the plasma is high. The level of EIC in the synovial fluids correlated positively with granulocyte count and-1-protease inhibitor (-1-PI) level, and this, too, tends to be higher when titer of RAHA in synovial fluid is high. The results suggested that the level of EIC in the plasma or synovial fluids can be a good marker for the systemic or localized activation of the granulocytes and that IgM rheumatoid factor (IgMRF) is involved in the mechanism of the release of elastase.     

What is the ability of anti-cyclic citrullinated peptide antibodies determination in synovial fluid in discriminating rheumatoid arthritis from non-rheumatoid arthritis patients? A Tunisian cross-sectional study

Anti-cyclic citrullinated peptide antibodies (ACPA) seem to be produced locally at the site of joints inflammation in the first stage of rheumatoid arthritis (RA). A strong correlation between serum ACPA and ACPA in the synovial fluid (SF-ACPA) is now suggested. A case-control study was conducted to evaluate the usefulness of ACPA determination in SF of patients with RA. A total of 53 patients with a knee-joint effusion (26 RA, 18 peripheral spondyloarthropathies (SPA), and 9 osteoarthritis (OA)) were included in our study. SF samples were obtained by performing therapeutic arthrosynthesis. IgG serum ACPA and SF-ACPA levels were determined by the enzyme-linked immunosorbent assay (ELISA). We have also determined IgG levels in serum and SF by nephelometry. Higher levels of IgG ACPA antibodies in SF (p = 0.045) and serum (p = 0.045) were found in patients with RA with respect to SPA and OA patients. The Spearman correlation analysis showed a significant and positive correlation between ACPA in serum and SF (rho = 0.516; p = 0.007) not only in the RA group but also in patients with SPA. Serum ACPA discriminated RA from non-RA at a cut-off value of 2.7 U/ml (sensitivity, 69%; specificity, 78%; and area under the curve (AUC), 0.72), whereas SF-ACPA discriminated RA from non-RA at a higher cut-off value of 4.95 U/ml (sensitivity, 73%; specificity, 61%; and AUC, 0.71). Our study suggests that the determination of SF-ACPA give complement information to serum ACPA in patients with RA.     

Increased levels of thymosin β4 in synovial fluid of patients with rheumatoid arthritis: association of thymosin β4 with other factors that are involved in inflammation and bone erosion in joints

Aim: Thymosin (Tβ4) may have various biological effects that are relevant to the pathogenesis of rheumatoid arthritis (RA). This study was performed to gain insight into the relevance of Tβ4 in the pathogenesis of inflammatory arthritis. Method: The level of Tβ4 in synovial fluid from patients with osteoarthritis (OA) or RA was measured by enzyme‐linked immunosorbent assay. An association between Tβ4 and matrix metalloproteinase (MMP)‐1 and MMP‐13 (collagenases), MMP‐2 and MMP‐9 (gelatinases), MMP‐7, adiponectin, lactoferrin, vascular endothelial growth factor (VEGF), urokinase‐type plasminogen activator (uPA), interleukin (IL)‐6, IL‐8 and prostaglandin E2 (PGE₂) in synovial joint fluids from OA and RA patients were investigated. Results: The level of Tβ4 in the synovial joint fluid of patients with OA and RA was (mean ± SD) 145 ± 88 and 1359 ± 1685 ng/mL, respectively. The level of Tβ4 in the synovial joint fluid of RA patients was significantly associated with the levels of MMP‐9, MMP‐13, VEGF, uPA, IL‐6 and IL‐8, but not with MMP‐1, MMP‐2, MMP‐7, adiponectin and lactoferrin. In contrast, the level of Tβ4 in the synovial joint fluid of patients with OA was not associated with any of these molecules. Conclusions: The results suggest that Tβ4 may play an important role in bone degradation and inflammation in RA but not OA, although nothing is known about the molecular mechanisms mediating Tβ4 in arthritic joints. The role of Tβ4 in arthritis should be studied to understand its relevance to the pathogenic processes in arthritis.     

Preliminary observation on the activity of T lymphocyte α-naphthyl acetate esterase (ANAE) in blood of patients with schistosomiasis japonica.

31 blood samples from proven patients with chronic schistosomiasis, 35 from latestage patients, and 37 from healthy persons were tested by both T lymphocyte esterasestainina method and lymphocyte transformation test (LTT). Both ANAE-staining posi-     

Anti-p53 antibodies are rarely detected in serum of patients with rheumatoid arthritis and Sj?gren's syndrome.

OBJECTIVE: To detect evidence of abnormalities of the p53 protein in autoimmune diseases. Mutation of the p53 protein may inhibit apoptosis and thereby lead to cancer and possibly play a role in the pathogenesis of autoimmune diseases. METHODS: Serum antibodies to p53 are detected in 30 to 50% of patients with cancer who have p53 mutations. Using an ELISA, we determined the prevalence of anti-p53 antibodies in the serum of 106 patients with rheumatoid arthritis (RA), 72 patients with primary Sj?gren's syndrome (SS), and 14 patients with lymphoma complicating SS. The presence of anti-p53 antibodies was also measured in the synovial fluid of 16 patients with RA. Positive sera by ELISA were confirmed by immunoprecipitation. RESULTS: Serum anti-p53 antibodies were detected in 2 of 106 patients with RA. The synovial fluid of one of these 2 patients was also studied and was positive. Anti-p53 antibodies were not detected in the other synovial fluids. Serum anti-p53 antibodies were not detected in 72 patients with primary SS alone, but were present in 2 of 14 patients with lymphoma complicating SS. CONCLUSION: Our results suggest that if p53 mutations have any role in the pathogenesis of some autoimmune diseases, they are rarely associated with the presence of anti-p53 antibodies in patients with RA. In patients with SS, the presence of serum anti-p53 antibodies might be an indirect sign of the development of a lymphoma.     

Increased levels of stromelysin-1 and tissue inhibitor of metalloproteinases–1 in sera from patients with rheumatoid arthritis

Objective. To evaluate the efficacy of stromelysin-1 (matrix metalloproteinase–3 [MMP-3]) and tissue inhibitor of metalloproteinases–1 (TIMP-1) in serum as markers for joint inflammation in rheumatoid arthritis (RA). Methods. Levels of both macromolecules in sera from 97 healthy controls, 109 patients with RA, and 47 patients with osteoarthritis (OA) were measured by respective 1-step sandwich enzyme immunoassays. In the patients with RA, serum levels of MMP-3 and TIMP-1 were investigated in relation to laboratory and clinical measures of disease activity. In addition, the relationships between serum and synovial fluid (SF) levels in paired samples from individual patients were examined. Results. Serum levels of both MMP-3 and TIMP-1 in RA patients were significantly higher than those in OA patients and in healthy controls (P < 0.001), and were shown to correlate with traditional systemic markers of inflammation including the erythrocyte sedimentation rate and C-reactive protein level, and with the Lansbury articular index. In addition, it was noted that serum levels of MMP-3 correlated with the corresponding values in paired SF samples obtained concurrently from patients with RA (r_s = 0.588, P < 0.001), while such correlations were not found for TIMP-1 levels. Conclusion. Our results support the notion that levels of both MMP-3 and TIMP-1 in RA patient sera are increased in association with inflammation. Furthermore, the level of MMP-3 in serum provides a particularly useful marker of inflammatory activity in the joints of patients with RA.     

Disease activity and the course of elbow joint deterioration over 10 years in the patients with early rheumatoid arthritis

One hundred and eighteen elbows in 59 patients who started treatment using disease-modifying antirheumatic drugs (DMARDs) in the first year of the disease were followed-up for more than 10 years without biologic agents. Using annual radiographs of the elbow joint, Larsen grade (LG) was determined, and bone absorption ratio (BAR), cortical thickness ratio (CTR) in the humerus, and olecranon thickness ratio (OTR) were calculated. Disease activity was determined by disease activity score (DAS)28-C-reactive protein (CRP)(3) at 3- or 4-month interval throughout the follow-up period. At 10 years, 30 elbows were in LG III or more, the more deteriorated (MD) group, and 88 elbows were in LG II or less, the less deteriorated (LD) group. In the radiological assessment, the mean LG, BAR, CTR, and OTR progressed with time linearly during 10 years. In the comparison between the MD group and the LD group, there was a significant difference in the magnitude of change in CTR (DeltaCTR; P = 0.0064), BAR (DeltaBAR; P = 0.0100), and OTR (DeltaOTR; P = 0.0051). There was a significant difference in the mean DAS28-CRP(3) (0-2 and 0-10 years) between the two groups (P = 0.0017 and 0.00002). The cut-off value of mean DAS28-CRP(3) (0-2 years), which indicated further progress to the MD group at 10 years, was 3.15. It is important to keep disease activity in low level to prevent progression of the elbow joint deterioration in the patients with RA.     

Do biologic drugs affect the need for and outcome of joint replacements in patients with rheumatoid arthritis? A register-based study

Objectives

The aim was to study the incidence of joint replacements among biologic drug and disease-modifying anti-rheumatic drug (DMARD) users as well as to investigate the plausible effect of biologic treatment on survival of prostheses in patients with Rheumatoid arthritis (RA).

Methods

The study population comprised 2 cohorts of patients [Register of biologic treatment in Finland (ROB-FIN) and the Central Finland RA database] from 1999 to 2010. Records of joint replacements performed in the study population between 1980 and 2010 were retrieved from the Finnish Arthroplasty Register. Propensity score matching was used to equalize patient characteristics between biologics and DMARD users. The incidence rates of primary and revision operations were compared between the 2 treatment groups. Kaplan–Meier survival analysis was used to analyze prosthesis survival.

Results

Of the 2102 biologics and 2710 DMARD users identified from the registries, 1587 were included in both groups after the matching. Median follow-up times were 3.1 and 8.0 years, respectively. There were more primary operations per 100 patient years in the biologics (3.89, CI 95% 3.41–4.41) vs. DMARD (2.63, 2.35–2.94) group but slightly fewer revisions (0.65, 0.46–0.88 vs. 0.83, 0.68–1.01). Biologics users were more likely to receive a joint replacement to small joints (p < 0.001). The survival of the prostheses installed during or prior to follow-up was similar in both treatment groups.

Conclusions

The use of biologic drugs did not reduce the need for joint replacement surgery in patients with a similar on-medication disease activity. Despite possibly lower rate of revisions among biologic users, the durability of prostheses was not improved.     

Different ELR (+) angiogenic CXC chemokine profiles in synovial fluid of patients with Behçet’s disease, familial Mediterranean fever, rheumatoid arthritis, and osteoarthritis

The aim of the present study was to determine synovial levels of ELR (+) CXC chemokines, known to attract mainly neutrophils to inflamed tissues by binding the neutrophil chemokine receptors CXCR1 and CXCR2 and promoting neovascularization in patients with various inflammatory disorders. The study group consisted of 14 patients with Behçets disease and nine with familial Mediterranean fever. Fourteen patients with rheumatoid arthritis and 16 with osteoarthritis served as controls. Synovial chemokine levels were measured by two-step sandwich enzyme-linked immunosorbent assay, and significant differences were found in the various chemokines studied. In addition to its angiogenic properties, increased synovial levels of interleukin-8 by attraction of more neutrophils to synovial fluids might also be responsible for the acute synovitis in patients with Behçets disease. However, the absence of chronic changes with the eventual development of pannus and erosions might result from relatively lower expression of interleukin-8 and the transient, short-lived nature of the arthritis observed in these patients.     

High frequency of vertebral fracture and low bone quality in patients with rheumatoid arthritis—Results from TOMORROW study

Objectives: Osteoporosis is one of the complications in patients with rheumatoid arthritis (RA). In this study, we researched the morbidity of existing vertebral fractures and the risk factors for vertebral fractures in patients with RA.

Methods: This study included 413 participants, 208 patients with RA, and 205 age- and sex-matched controls without RA. Clinical data, radiographic assessment of vertebral fracture from T4 to L4 in thoracic and lumber spine, bone mineral density (BMD), and bone metabolic markers (BMM) were analyzed.

Results: Vertebral fractures were observed more frequently, severe and multiple in patients with RA. In the logistic regression analysis, age (adjusted odds ratios (OR): 1.07, 95% confidence interval (CI): 1.04–1.09) and RA (adjusted OR: 1.72, 95% CI: 1.04–2.83) were risk factors for existing vertebral fracture. Moreover, two bone matrix-related markers, undercarboxylated osteocalcin (ucOC) (adjusted OR: 1.68, 95% CI: 1.02–2.78), and urinary pentocidine (adjusted OR: 2.51, 95% CI: 1.48–4.24) were associated with existing vertebral fracture.

Conclusions: High frequent, multiple, and severe vertebral fractures were found in patients with RA compared to the controls. Low bone quality might be the cause of the frequent prevalence of vertebral fracture in patients with RA.     

An elevated level of IL-10- and TGFβ-secreting T cells,B cells and macrophages in the synovial membrane of patients with reactive arthritis compared to rheumatoid arthritis

A relative high secretion level of IL-10 and a low secretion of TNF- has been described in the synovial fluid and peripheral blood of patients with reactive arthritis (ReA), possibly contributing to the persistence of bacteria. The role of TGF- is less clear. We investigated these cytokines in the synovial membrane of patients with ReA and rheumatoid arthritis (RA) and tried to identify their cellular source. We used sections from the synovial membrane of 4 ReA and 4 RA patients which were double stained with immunofluorescence antibodies against cell surface markers for T cells (CD3), macrophages (CD68) and B cells (CD20) in combination with antibodies against intracellular cytokines TNF-, IFN-, TGF-, IL-4 and IL-10, and quantified these using a fluorescence microscope. A lower number of TNF--secreting cells were found in ReA compared to RA: CD3+: 1.78±0.54% versus 5.02%±0.47% (p=0.034). CD68+: 2.86±0.52 versus 5.37±0.53% (p=0.034), CD20+ : 3.02±0.42% versus 3.58±0.48% (p>0.05). A higher number of IL-10 positive cells were found in ReA compared to RA: CD3+: 3.27±1.5% versus 1.13±0.50% (p=0.034), CD68+ 1.23±0.75% versus 0.83±0.35%(p>0.05), CD20+: 3.70±1.6% versus 1.6±1.1% (p>0.05). A difference between ReA and RA was also found for TGF-+ T cells: CD3+ 7.86+1.5% versus 1.78+0.35% (p= 0.032); CD20+: 7.91+2.1% versus 2.1+2.8% (p>0.05), CD68+: 7.81%+1.24% versus 2.12+0.28% (p= 0.032). In conclusion, we saw a different cytokine secretion pattern in the synovial membrane of ReA and RA. For T cells in ReA we found a cytokine secretion profile typical for T regulatory cells 1 (Tr1), with an elevated level of IL-10- and TGF--secreting cells. Whether this is due to a more general difference in TNF-, IL-10 or TGF- production which is genetically determined or regulated by T cells remains to be determined.Abbreviations AS Ankylosing spondylitis - RA Rheumatoid arthritis - ReA Reactive arthritis     

Anti-cyclic citrullinated peptide antibodies do not reflect self-reported disability and physical health in patients with rheumatoid arthritis of less than 5 years of duration

It is well accepted that patients with antibodies against cyclic citrullinated peptides (anti-CCP) and rheumatoid arthritis (RA) suffer from more severe forms of RA in terms of clinical presentation and radiographic destruction at long term compared to anti-CCP-negative patients. The purpose of this cross-sectional study was to investigate whether the measures of self-reported health among patients with RA of <5 years of duration are influenced by anti-CCP status. Additionally, we aimed to determine whether the measures of self-reported health among the two patient groups differ from those of a control group. Telephone interviews were conducted with 464 patients with RA and 637 population controls, who reported educational level, income, smoking habits and lifestyle 10 years before the interview and completed the Health Assessment Questionnaire and the Short-Form Health Survey Questionnaire, version 2 (SF-36v2); 424 (91 %) patients submitted a blood sample for analysis. Patients with anti-CCP-positive and anti-CCP-negative RA showed no significant differences in self-reported disability and physical health after adjustment for age, gender, socioeconomic factors, lifestyle and disease-related variables (p > 0.05). Both groups of RA patients reported significantly more physical disabilities in everyday life and significantly poorer physical health than the controls (both p < 0.001). A similar pattern was seen for self-reported mental health (both p < 0.05). Patients with RA of <5 years of duration report significantly more disability and poorer physical health than the general population of Denmark, but these reports were independent of anti-CCP status.     

Sustained elevation of interleukin-33 in sera and synovial fluids from patients with rheumatoid arthritis non-responsive to anti-tumor necrosis factor: possible association with persistent IL-1β signaling and a poor clinical response

Although TNF inhibitors have dramatically improved the outcome of patients with rheumatoid arthritis, 30–40% of patients do not respond well to them and treatment needs to be changed. In an effort to discriminate good and poor responders, we focused on the change in serum and synovial fluid levels of interleukin (IL-) 33 before and after treatment with TNF inhibitors. They were also measured in synovial fluids from 17 TNF inhibitor-na?ve patients, and fibroblast-like synoviocytes (FLS) in-culture from 6 patients and correlated with various pro-inflammatory cytokines. Serum levels of IL-33 at 6?months after treatment decreased significantly in responders, while they did not change in non-responders. Synovial fluid levels of IL-33 in 6 patients under treatment with TNF inhibitors stayed high in 3 who were refractory and slightly elevated in 2 moderate responders, while they were undetectable in one patient under remission. Among inflammatory cytokines measured in 17 synovial fluids from TNF inhibitor-na?ve patients, levels of IL-33 showed a significant positive correlation only to those of IL-1β. IL-1β increased IL-33 expression markedly in FLS in vitro, compared to TNF-α. IL-1β might be inducing RA inflammation through producing pro-inflammatory IL-33 in TNF inhibitor-hypo-responders. Sustained elevation of serum and/or synovial levels of IL-33 may account for a poor response to TNF inhibitors, although how TNF inhibitors affect the level of IL-33 remains to be elucidated.     

Peripheral blood and synovial fluid monocyte activation in inflammatory arthritis. i. a cytofluorographic study of monocyte differentiation antigens and class ii antigens and their regulation by γgm-interferon

Recent study of the expression of monocyte differentiation antigens (MAg) and HLA–DR on peripheral blood monocytes (PBM) has led to the recognition of resting and activated monocyte phenotypes. The former is identified by the expression of large amounts of MAg (i.e., Mo2 and 63D3) and small amounts of HLA–DR, while the latter is identified by the reverse. We studied the phenotypes of PBM and synovial fluid monocytes (SFM) of patients with chronic inflammatory arthritis and found that PBM were primarily resting and SFM were usually activated. In addition, we measured the degree of modulation of MAg and HLA–DR by γ-interferon (γ-IFN). Patient PBM reacted the same as PBM from normal individuals (i.e., MAg decreased and HLA–DR increased after exposure to γ-IFN). However, in patient SFM, HLA–DR did not increase with exposure to γ-IFN because expression was already maximal. Interestingly, MAg could still be down-regulated on γ-IFN–treated SFM, even when expression began at a very low level (i.e., activated phenotype). This independent regulation of MAg and HLA–DR suggests that macrophage activating factors other than γ-IFN may be responsible, in part, for the activated phenotypes observed.