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PubMedDate not supplied

Vascular endothelial growth factor levels in the serum and synovial fluid of patients with rheumatoid arthritis.

OBJECTIVE: To determine the vascular endothelial growth factor (VEGF) concentrations in serum and synovial fluid (SF) from patients with rheumatoid arthritis (RA) and to search for relationships between VEGF levels and clinical and laboratory variables. METHODS: We measured VEGF levels using an enzyme-linked immunosorbent assay. Serum samples were obtained from 99 RA patients, 49 osteoarthritis (OA) patients, and 80 normal controls. Paired samples of serum and SF were collected from 32 patients with RA and 15 with OA. RESULTS: The mean serum VEGF concentration was 590.1 pg/ml for RA patients, 286.7 pg/ml for OA patients, and 265.8 pg/ml in controls. The serum VEGF concentration was significantly higher in the RA patients than in the OA patients or the controls (both p < 0.001). Furthermore, the VEGF levels in SF from RA patients were significantly higher than in SF from OA patients (p = 0.017). However, there was no correlation between VEGF levels in serum and SF from the same RA patients. The serum VEGF concentration was correlated with the ESR, serum CRP concentration, serum rheumatoid factor, number of tender and swollen joints, Modified Health Assessment Questionnaire, and patient and physician global assessments of disease activity in RA patients. CONCLUSION: These results suggest that VEGF level is related to RA disease activity, suggesting that VEGF may play some role in the pathogenesis of RA.

Adult↗
PubMed1995

Neutrophil gelatinase levels in plasma and synovial fluid of patients with rheumatic diseases.

To examine the clinical significance of neutrophil gelatinase in rheumatic diseases, plasma and synovial fluid (SF) gelatinase levels were determined in 62 patients with rheumatoid arthritis (RA), 12 patients with ankylosing spondylitis (AS), 18 patients with osteoarthritis (OA) and 17 healthy controls. The gelatinase level was measured by enzyme-linked immunoassay (ELISA). The assay had a sensitivity of 1 ng/ml and a working range of 5-25 ng/ml. Gelatinase levels were significantly higher in the plasma of patients with RA and of patients with RA complicated by amyloidosis or vasculitis as compared to those of healthy controls. Moreover, the mean value of gelatinase in the plasma of patients with RA complicated by vasculitis was found to be significantly higher than that of RA patients without vasculitis. A significant increase in gelatinase concentration was also observed in the plasma of AS patients but not in the plasma of patients with OA. The concentration of gelatinase in the RA SF samples was much higher (18-fold) than the level of the enzyme in the plasma of RA patients. There was also a higher concentration of gelatinase (four-fold) in OA SF compared with OA plasma. The results suggested that circulating gelatinase may reflect some degree of neutrophil activation in patients with inflammatory arthritis, especially in those with RA complicated by vasculitis. However, the results did not allow a differentiation between chronic and acute inflammation.

Adult↗
PubMedDate not supplied

An ultrastructural study of the synovial fluid (S.F.) in rheumatoid arthritis versus ankylotic spondylarthritis.

Our studies focused on the cytomorphological and ultrastructural analysis of ten synovial fluid (S.F.) samples from patients with serum positive rheumatoid arthritis (R.A.) (latex 1/1280, 1/640 and Waaler-Rose 1/1024, 1/1512) as compared to five S.F. sampes from patients with peripheral ankylotic spondylarthritis (A.S.). The cytomorphological investigation aimed at defining the cellularity, ragocytosis and synoviocytogram. We found out that the average number of cells (R.A. = 8060/mm3; A.S. = 6100/mm3), percentage of ragocytes (R.A. = 75%; A.S. = 25%), polymorphonuclear cells (R.A. = 70%; A.S. = 58%), degradative polymorphonuclear cells (R.A. = 7%, A.S. = 3%) and phagocytic mononuclear cells (R.A. = 16%; A.S. = 13%) are by far larger in R.A. than in A.S., whereas the lymphocyte percentage is much more reduced (R.A. = 13%; A.S. = 26%). Ultrastructurally, the rheumatoid S.F. ragocytes present specific intracytoplasmic inclusions and phagolysosomes pointing to an endocytotic activity. At the level of the degradative polymorphonuclear cells (7% cytomorphologically confirmed), the alteration of the granular-fibrillar structure of the nucleus and the presence of some cytoplasmic lysis areas were associated with the absence of cellular organelles. We also noticed cellular and collagen detritus. In A.S. the ultrastructural effect on the polymorphonuclear cells is much more reduced (3%) as compared to R.A. and resides in the dilatation of both mitochondrial cristae and granularendoreticular cisternae as well as in a slight concentration of the fibrogranular nuclear matter. The cells are considerably active in endocytosis. The phagocytotic mononuclear cells of the rheumatoid S.F. and A.S. are morphologically identical to the immunologically activated macrophages.(ABSTRACT TRUNCATED AT 250 WORDS)

Arthritis, Rheumatoid↗
PubMed1998

T cell derived cytokines in psoriatic arthritis synovial fluids.

OBJECTIVE: The aim of this study was to investigate the concentrations of T cell derived cytokines in the synovial fluids (SFs) of patients with psoriatic arthritis (PsA) in comparison with rheumatoid arthritis (RA) and osteoarthritis (OA). METHODS: Th1 type cytokines (interleukin 2 (IL2), tumour necrosis factor beta (TNF beta), and interferon gamma (INF gamma) and Th2 type cytokines (IL4, IL10) were measured by means of enzyme linked immunosorbent assays. RESULTS: IL2 was usually not detectable in any of the disease groups. TNF beta was found in 3 of 31 PsA SFs (mean (SEM) 11.1 (2.3) pg/ml) and in a significantly lower concentration than in 20 of the 40 RA SFs (42.2 (15.6) pg/ml; p < 0.002). INF gamma was measurable in 2 of 10 PsA and 6 of 16 RA SFs (p > 0.05). IL4 was present at low concentrations in 4 of 22 PsA SFs (0.41 (0.8) pg/ml), and in 15 of 20 RA SFs (0.63 (0.09) pg/ml; p < 0.01). IL10 was found in 4 of 27 PsA SFs (12.3 (0.9) pg/ml) and in 27 of 32 RA SFs (37.3 (4.9) pg/ml; p < 0.0001). In all OA SFs cytokine concentrations were below the limit of detection. CONCLUSION: The pattern of T cell derived cytokines in PsA SFs was similar to that of RA SFs. However, both the frequency and the concentrations of cytokines were lower in PsA SFs than in RA SFs, while OA SFs generally lacked any detectable T cell cytokines altogether. The presence of Th1 and Th2 cell derived cytokines in PsA SFs suggests the presence of activated T cells in the inflamed joint tissues and their participation in the immunoinflammatory events.

Arthritis, Psoriatic↗
PubMed2002

Interleukin 13 in synovial fluid and serum of patients with psoriatic arthritis.

OBJECTIVES: To compare the pattern of interleukin (IL) 13 production in synovial fluid (SF) and serum of patients with psoriatic arthritis (PsA) with that in patients with rheumatoid arthritis (RA) and osteoarthritis (OA), investigating its relation to the proinflammatory cytokine IL12. METHODS: SF and serum IL13 levels were determined in 35 patients with PsA, 36 with RA, and 15 with OA. The main clinical and laboratory variables, including number of painful and/or swollen joints, Ritchie index, morning stiffness, erythrocyte sedimentation rate, level of C reactive protein, level of rheumatoid factor, and SF analysis, were also evaluated. RESULTS: SF IL13 levels were significantly higher in patients with PsA (p<0.02) or RA (p<0.012) than in patients with OA, with no significant difference between the former two. SF IL12 levels were significantly higher in patients with PsA (p<0.023) than in those with OA. Serum IL13 (p<0.0001) and IL12 (p<0.02) levels were lower in patients with PsA than in those affected by RA. Only patients with PsA had higher IL13 levels in SF than in serum (p<0.002). The IL13 SF/serum ratio was higher in the PsA group than in the group with RA (p<0.005) or OA (p<0.026). SF IL13 levels correlated with serum IL13 levels (p<0.0001) in RA and with SF IL12 levels (p<0.03) in PsA. CONCLUSIONS: In PsA, there appears to be localised production of IL13, in balance with IL12, in the inflamed joints. The distinct IL13 secretion profiles in PsA, RA, and OA may be related to the clinical pictures, reflecting the different pathogenic mechanisms involved in inflammatory and degenerative joint diseases.

Arthritis, Psoriatic↗
PubMed1998

Expression of macrophage markers by a population of T cells obtained from synovial fluid of a subgroup of patients with juvenile rheumatoid arthritis.

OBJECTIVE: To characterize distinctive lymphoid cell populations in the synovial fluid (SF) of patients with juvenile rheumatoid arthritis (JRA) that have the specific ability to display monocytic markers when cultured in vitro. METHODS: Mononuclear cells obtained from SF of patients with JRA and depleted of adherent macrophages were cultured in vitro in RPMI 1640 medium supplemented with only fetal calf serum (FCS). Phenotypic evaluation of these cells was by flow cytometry and immunohistochemical analysis was by specific fluorochrome labeled antibodies. RESULTS: T cells from a JRA subgroup displayed some typical macrophage attributes, i.e., abundant cytoplasm, adherence to plastic, and phagocytosis of latex beads when cultured in vitro. These cells have the ability to survive in culture for several weeks in RPMI 1640 medium containing only 10% FCS. The macrophage-like T cells rosetted with sheep red blood cells and proliferated when stimulated with phytohemagglutinin or anti-CD3, indicating functional T cell responses. CONCLUSION: Our data indicate that a population of T cells obtained from the SF of a subgroup of patients with JRA exhibited characteristics of macrophages, yet retained their CD3 and T cell receptor expression. Whether this promiscuous behavior is caused by malignant transformation of lymphoid precursor cells or is induced by the concerted effect of a myriad of cytokines and growth factors present in the SF remains unknown. The presence of these cells in the SF of 2 patients with JRA with different onset types raises the question of their function and significance in an autoimmune disorder such as JRA.

Adolescent↗
PubMed2000

Placenta growth factor (PlGF) induces vascular endothelial growth factor (VEGF) secretion from mononuclear cells and is co-expressed with VEGF in synovial fluid.

The aims of this study were (i) to determine whether PlGF, VEGF and PlGF/VEGF heterodimers are detected in synovial fluid (SF) and plasma samples from patients with a range of arthropathies; (ii) to describe whether any correlation exists between SF PlGF, VEGF and PlGF/VEGF heterodimer levels and the total and differential SF leucocyte counts; and (iii) to investigate the regulation of peripheral blood mononuclear cell (PBMC) VEGF secretion by stimuli relevant to inflammatory joints. PlGF, VEGF and PlGF/VEGF heterodimer levels were measured in the SF and plasma of patients with a range of arthropathies and normal controls by ELISA. Western blotting for PlGF was performed on SF from three patients with rheumatoid arthritis (RA) and primary inflammatory arthropathies. VEGF was quantified in cell culture supernatants after stimulation with lipopolysaccharide (LPS), PlGF or cobalt ions of PBMC isolated from RA patients and controls. PlGF and VEGF were detected in all SF samples. PlGF/VEGF heterodimers were detected in 10.2% of SF samples, most frequently in RA samples. Western blotting confirmed the presence of PlGF in RA SF. PlGF was detected in 52% of RA and 31% of control plasma samples, and VEGF was detected in 38% of RA and 38% of control plasma samples. PlGF/VEGF heterodimers were detected in 21% of RA samples and none of the control samples. In primary inflammatory arthropathy patients, SF PlGF and VEGF levels correlated significantly with the SF total leucocyte count and the neutrophil count. PlGF was the most potent inducer of PBMC VEGF production in both RA and control subjects. This is the first report of the detection of PlGF and PlGF/VEGF heterodimers in the SF of patients with inflammatory arthropathies, and we have shown for the first time that PlGF up-regulates PBMC VEGF production. PlGF may therefore play a key role in the production of VEGF in the inflammatory joint.

Adolescent↗
PubMed1980

Synovial fluid leukocytosis associated with intracellular lipid inclusions.

A unique case of an inflammatory monoarthritis associated with intracellular lipid inclusions in synovial fluid leukocytes is described. In contrast to previous reports, these lipid inclusions were uniformly intracellar, not associated with articular destruction, and not visible under ordinary light but readily seen under polarized light microscopy.

Arthritis↗
PubMed1981

Sodium oxacillin in the horse: serum, synovial fluid, peritoneal fluid, and urine concentrations after single-dose intramuscular administration.

Six healthy adult mares were given a single dose (25 mg/kg of body weight) of sodium oxacillin IM. Oxacillin concentrations in serum, synovial fluid, peritoneal fluid, and urine were measured serially over a 48-hour period. The mean peak serum oxacillin concentration was 9.75 microgram/ml at 0.5 hour after injection. Mean peak oxacillin concentrations in synovial and peritoneal fluids were 1.45 microgram/ml and 2.60 microgram/ml at 1 hour and 2 hours, respectively. These concentrations decreased in parallel with serum values and were not measurable at 48 hours. Urine concentrations of oxacillin were high, with a mean peak concentration of 2,790.2 microgram/ml at 0.5 hour.

Animals↗
PubMed2000

Detection and quantitation of stem cell factor (kit ligand) in the synovial fluid of patients with rheumatic disease.

OBJECTIVE: To identify and quantitate stem cell factor (SCF; kit ligand) in the serum and synovial fluid (SF) of patients with rheumatoid arthritis (RA) and to compare these values with those measured in normal serum, RA serum, SF of patients with other rheumatic diseases, and conditioned medium from cultured synoviocytes. METHODS: SCF was measured in serum, SF, and conditioned synovial cell culture medium by a sensitive ELISA. Results were correlated with hematologic variables including white blood cell count, hemoglobin, platelet count, erythrocyte sedimentation rate, and rheumatoid factor. RESULTS: SCF levels in RA SF exceeded those measured in RA serum, osteoarthritis SF, and SF from patients with other inflammatory arthropathies. SCF was detectable in conditioned medium from cultured synoviocytes. CONCLUSION: High levels of SCF are present in RA serum and SF. Local production of SF may influence expansion of myeloid progenitor cells and mast cell function in RA.

Arthritis, Rheumatoid↗
PubMed1991

Characterization of a gelatinase from human rheumatoid synovial fluid cells.

A metalloproteinase with a specificity for gelatin was isolated from serum-free medium of cultures of rheumatoid synovial fluid. The enzyme showed all the properties of a leukocyte gelatinase. In addition to gelatin this proteinase cleaved the synthetic substrate dinitrophenyl-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg (Dnp-peptide) rapidly, while casein was a much poorer substrate. This proteinase showed no enzymatic activity against collagen type I, was secreted in a latent form and could be activated by trypsin or organomercurial compounds, such as mersalylic acid or 4-aminophenyl-mercury acetate. The latent enzyme had an apparent molecular mass of 130,000-150,000 estimated by gel filtration or 97,000 by electrophoresis on polyacrylamide gel containing sodium dodecyl sulphate. When analysed by immunoblotting the enzyme was recognized by antibodies raised against human polymorphonuclear leukocyte gelatinase. Although we found synovial fibroblasts to be largely present in the cell cultures we could not detect any fibroblast gelatinase activity.

Arthritis, Rheumatoid↗
PubMed1985

Production of mononuclear cell factor by mononuclear phagocytes from rheumatoid synovial fluid.

Collagenase-and PGE1-stimulating activities (mononuclear cell factor or MCF) have been found in culture supernatants from synovial fluid macrophages (SF-M phi) of patients wih rheumatoid arthritis. The apparent molecular weight of the MCF activity present in SF-M phi supernatants was between 15-20,000 daltons, which is similar to interleukin I. All the SF-M phi supernatants from patients tested showed the MCF activity. Our study presents evidence for MCF production by monocyte-macrophages at the local site of the lesion.

Adult↗
PubMed1981

Cytochemical detection of proteases in synovial fluid of arthritic joints.

A cytochemical method can be used to detect neutral proteases in polymorphonuclear leukocytes (PMN cells) in synovial fluid smears obtained from arthritic joints. Using this method, incubation of fixed smears in NaCl-borate buffer causes release of neutral PMN proteases, the enzymatic activity of which id documented in the degradation of exudate protein in ring-shaped areas centered around the neutrophils(halo effect). Such halo formation can also be detected around certain large mononuclear cells, as well as around very small particles - which may represent thrombocytes. Application of trypsin inhibitors and various antirheumatic agents causes concentration-dependent inhibition of halo formation.

Arthritis, Rheumatoid↗
PubMed2000

Effect of IL15 on T cell clonality in vitro and in the synovial fluid of patients with rheumatoid arthritis.

OBJECTIVE: Recent studies have suggested that interleukin (IL) 15 induces T cell accumulation in synovial lesions of rheumatoid arthritis (RA). This study aimed at determining whether this cytokine could explain in vivo T cell clonality in RA. METHODS: Peripheral blood mononuclear cells (PBMC) from patients with RA were stimulated in vitro with IL15 or IL2. After isolation of mRNA from stimulated cells and synovial T cells, genes coding the V-D(N)-J (CDR3) region of T cell receptor beta chains were amplified by a reverse transcriptase polymerase chain reaction. A single strand conformation polymorphism analysis was used to detect the clonotype(s) of accumulating T cells. Nucleotide and amino acid sequencing was also performed. RESULTS: Stimulation of PBMC with IL15 resulted in oligoclonal expansion of T cells. However, IL15 induced clones from PBMC were mostly different from the dominantly expanding T cell clones in synovial fluid. Furthermore, IL15 and IL2 responding clones were only partially identical. CONCLUSIONS: Although IL15 results in clonal accumulation of T cells, T cell clonality in rheumatoid joints could not be explained by the effect of IL15 alone. The results indicated the requirement of other factor(s), in addition to IL15, in the pathological process affecting RA joints. The results also suggested different responses by each T cell clone to IL15 or IL2.

Aged↗
PubMed2005

Examination of the molecular nature of low-molecular-mass chromium(III) ions in isolated osteoarthritic knee-joint synovial fluid.

High field (1)H NMR spectroscopy demonstrated that equilibration of added Cr(III) ions in osteoarthritic knee-joint synovial fluid (SF) resulted in its complexation by a range of biomolecules, the relative efficacies of these complexants/chelators being threonine approximately alanine>glycine>glutamine>citrate>histidine approximately phenylalanine approximately tyrosine>valine approximately isoleucine approximately leucine>glutamate>lactate approximately acetate approximately formate approximately pyruvate, this order reflecting the ability of these ligands to compete for the available Cr(III) in terms of (1) thermodynamic equilibrium constants for the formation of their complexes and (2) their SF concentrations. The significance of these observations with regard to the in vivo corrosion of chromium-containing metal alloy joint prostheses (e.g., CoCr alloys) is discussed.

Cations↗
PubMed1989

Purification and characterization of extracellular phospholipase A2 from human synovial fluid in rheumatoid arthritis.

Extracellular phospholipase A2 was purified about 1.7 X 10(5) fold to near homogeneity from human synovial fluid of rheumatoid arthritis by sequential use of column chromatographies on heparin-Sepharose, butyl-Toyopearl, and reversed-phase HPLC. The final preparation showed a single band on SDS-polyacrylamide gel electrophoresis, and its molecular mass was estimated to be approximately 13,700 daltons. The purified enzyme had a pH optimum of 9.0 and required Ca2+ for maximum activity. It hydrolyzed phosphatidyl-ethanolamine more effectively than phosphatidylserine and phosphatidylcholine. These properties were similar to those of an extracellular phospholipase A2 detected in the peritoneal cavity of caseinate-treated rats.

Arthritis, Rheumatoid↗