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PubMed1983

Relationship of angiogenesis factor in synovial fluid to various joint diseases.

A low-molecular-weight freely dialysable angiogenesis factor has been isolated from 49 synovial fluids obtained from patients with various joint diseases. An analysis of disease type and incidence of freely dialysable angiogenesis activity showed that the osteoarthrotic group had a significantly higher incidence than all the other groups (p = 0.0332). Angiogenesis factor has also been detected in a bound form in the retentates of fluids which gave positive results for dialysable factor. The possibility that an imbalance between carrier-bound and free factor may have a causative role in disease is discussed.

Angiogenesis Inducing Agents↗
PubMed1995

Effects of synovial fluid hyaluronan concentration and molecular size on clearance of protein from the canine knee.

OBJECTIVE: The synovial fluid (SF) concentration of proteins (or fragments thereof) derived from the articular cartilage may reflect the processes of cartilage breakdown or repair within an arthritic joint and, hence, serve as a marker of these processes. Because changes in the clearance kinetics of marker molecules in SF would invalidate the assumption that their concentration in SF is directly proportional to turnover of the cartilage matrix, we investigated whether reduction in the concentration and/or molecular size of hyaluronan (HA) in joint effusions affects clearance of albumin from the canine knee. METHODS: The volume of distribution and clearance kinetics of radioiodinated serum albumin (RISA) were determined in normal dogs after injection of the knee with HA preparations whose concentration (3.0-0.03 mg/ml) and average molecular weight (0.2-6.1 x 10(6) Da) resembled those seen in synovial effusions, and which increased by roughly 270% the volume of fluid in the joint space. RESULTS: In knees injected with a physiologic concentration (3.0 mg/ml) of high molecular weight HA, the RISA distribution volume (2.8 ml) and clearance rate (1.7 microliters/min) slightly exceeded those in knees in which the SF HA concentration was reduced 55-62% by injection of 0.3-0.03 mg/ml HA, but not those in knees in which the average molecular weight of the SF HA was reduced 84% by injection of exogenous HA with a molecular weight of 2.4 x 10(5) Da. CONCLUSION: Our data suggest that SF HA is a relatively minor determinant of the kinetics of RISA clearance from the joint, and the changes in HA content seen in joint effusions are unlikely to confound interpretation of the concentration of marker molecules in SF.

Animals↗
PubMed1996

Identification of 2 Chlamydia trachomatis antigens recognized by synovial fluid T cells from patients with Chlamydia induced reactive arthritis.

OBJECTIVE: To identify antigens of Chlamydia trachomatis recognized by synovial T cell clones from patients with reactive arthritis (ReA). METHODS: C. trachomatis specific T cell clones were isolated from synovial fluid of 2 patients with chlamydia induced ReA. The particular antigens/epitopes recognized by these clones were identified using T cell immunoblotting and testing recombinant chlamydial proteins and synthetic peptides. RESULTS: Two sets of clones were shown by immunoblotting to recognize antigens of roughly 60 and 18 kilodaltons (kDa) respectively. By testing recombinant chlamydial proteins these antigens were identified as the 57 kDa heat shock protein and the 18 kDa histone-like protein, Hc1. Mapping the epitope in Hc1 using synthetic peptides identified a peptide containing a sequence motif compatible with binding to HLA-DR1, the restricting antigen for the Hc1 specific clones. CONCLUSION: These are the first 2 chlamydial antigens to be identified as targets of the synovial T cell response in chlamydia induced ReA. Both have properties that are shared with target antigens identified in ReA induced by enteric infection and relevant to the pathogenesis of joint inflammation.

Adult↗
PubMed2001

HPLC determination of adenosine in human synovial fluid.

A high-performance liquid chromatographic method has been developed for the quantitative determination of adenosine in human synovial fluid. The method is simple, rapid and, overall, selective. No interference with the components of the biological matrix was observed in these chromatographic conditions. An ODS (250 x 4.6 mm) 5 microm column was used with an isocratic elution of a phosphate buffer-acetonitrile mobile phase. Detection was carried out on a UV detector at 260 nm. Calibration curve was found to be linear in the 0.7--70 microg ml(-1) range. Linear regression analysis of the data demonstrates the efficacy of the method in terms of precision and accuracy. The precision of this method, calculated as the relative standard deviation (RSD) of the recoveries (1.57--2.21%), was excellent. The limits of quantitation (LOQ) and detection (LOD) were respectively 0.7 and 0.2 microg ml(-1). The method was applied to some samples of synovial effusion from patients affected by rheumatoid arthritis. The concentrations of adenosine which were found were included in the range of the calibration curve.

Adenosine↗
PubMed2003

Increased expression of alpha(1,3)-fucosyltransferase-VII and P-selectin binding of synovial fluid T cells in juvenile idiopathic arthritis.

OBJECTIVE: The mechanisms controlling the recruitment of T helper type 1 (Th1) cells to the inflamed synovium are not fully understood. Here, we focus on alpha(1,3)-fucosyltransferase-VII (FucT-VII), an enzyme responsible for the generation of functional P- and E-selectin ligands that is upregulated in Th1 cells. METHODS: Expression of transcripts encoding FucT-VII, interferon-gamma (IFN-gamma), and interleukin 12Rbeta2 (IL-12Rbeta2) were analyzed in T cells purified from synovial fluid (SF) and from peripheral blood (PB) of children with juvenile idiopathic arthritis (JIA) using kinetic reverse transcriptase polymerase chain reaction analysis. Binding of SF and PB T cells to P-selectin was determined by flow cytometry using a soluble P-selectin/IgG1 fusion molecule. Recruitment of T cells to synovial tissue in vivo was studied by analyzing the migration of FucT-VII transfected Jurkat T cells into human rheumatoid synovial tissue grafted into SCID mice. RESULTS: In patients with JIA, the mRNA levels of FucT-VII, as well as of IFN-gamma and IL-12Rbeta2, were up-regulated in SF T cells compared to paired PB T cells. A higher expression of FucT-VII mRNA in SF T cells was associated with increased binding of T cells to P-selectin. Moreover, FucT-VII expression and increased P-selectin binding capacity of T cells were associated with a polyarticular course of oligoarticular JIA. Expression of FucT-VII in Jurkat T cells resulted in an increased accumulation of these cells in human rheumatoid synovial tissue grafted into SCID mice. CONCLUSION: Our data indicate an important role of FucT-VII in the enhanced homing of T cells to the inflamed synovium.

Adolescent↗
PubMed1988

The clinical and immunological effects of pulse methylprednisolone therapy in rheumatoid arthritis. III. Effects on immune and inflammatory indices in synovial fluid.

Pulse methylprednisolone therapy has dramatic effects on cellular traffic and immunological variables of disease activity in both the synovial fluid (SF) and peripheral blood compartments. While these effects persisted at 2 weeks in the SF they had reverted to pretreatment levels in the peripheral blood. Pulse therapy resulted in temporary changes in the number of helper and suppressor cells in peripheral blood with an initial fall followed by a rise in the helper to suppressor ratio. In the SF, pulse therapy produced more gradual and sustained effects on the helper and suppressor subpopulations, resulting in a reduction in the helper to suppressor ratio.

Antigen-Antibody Complex↗
PubMed1989

Biochemical characterization of soluble phospholipase A2 from rheumatoid synovial fluid.

Radiolabeled E. coli, Phosphatidylethanolamine (PE) and Phosphatidylcholine (PC), were used to characterize the phospholipase A2 (PLA2) activity in synovial fluid (SF) from rheumatoid arthritis (RA) patients. Cell-free fractions of SF contain a PLA2 enzyme that preferentially releases [14C]oleic acid from E. coli, requires calcium and is optimally active at neutral pH. Purified PE, but not PC is also readily degraded by the soluble enzyme. A cell-associated PLA2 present in sonicates of SF mononuclear cells and neutrophils preferentially releases [3H]AA from E. coli. These studies suggest the presence of at least two different enzymes with activity of PLA2 in rheumatoid SF.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed1990

Development and evaluation of a method for preservation of synovial fluid wet preparations for quality control testing of crystal identification.

We describe a technique for embedding drops of fresh synovial fluid in mounting resin to prepare test slides that retain crystal morphology for at least 2 weeks. Hospital and private rheumatologists were solicited nationwide to test themselves and their staff with 5 unknown specimens. Eighty-nine percent identified monosodium urate (MSU) crystals from a tophus but only 71% identified smaller and less frequent MSU. Seventy-five percent correctly identified calcium pyrophosphate dehydrate and 42% detected steroid crystals. Apatite crystal clumps included as control slides with no birefringent material were suspected by only 7%. This or another method which simulates actual practice by also testing the equipment and handling of the microscope is proposed as an important component of quality control testing programs for synovianalysis.

Birefringence↗
PubMed2001

Correlations between matrix metalloproteinase-9 and adenosine deaminase isozymes in synovial fluid from patients with rheumatoid arthritis.

OBJECTIVE: To clarify the significance of increased adenosine deaminase (ADA) isozyme activities in synovial fluid (SF) from patients with rheumatoid arthritis (RA). METHODS: ADA isozyme activities were measured using colorimetric assays. Concentrations of matrix metalloproteinase-9 (MMP-9) were measured by ELISA. RESULTS: Levels of ADA isozyme activities in RA SF were significantly higher than those from patients with osteoarthritis and patients with traumatic injuries. Significant positive correlations between MMP-9 concentration and ADA activities were observed in RA SF (MMP-9 vs total ADA: r = 0.880; vs ADA1: r = 0.829; vs ADA2: r = 0.823; p < 0.001). CONCLUSION: Elevated levels of ADA activities were found in SF from patients with RA. There were significant positive correlations between MMP-9 and ADA isozymes. These results may reflect the inflammatory condition of patients with RA.

Adenosine Deaminase↗
PubMed1989

Iron-binding proteins and free iron in synovial fluids of rheumatoid arthritis patients.

Iron-binding proteins (lactoferrin, transferrin and ferritin) and free iron were measured in synovial fluid (SF) from 30 patients with rheumatoid arthritis (RA) and 20 osteoarthritis (OA) patients. The iron-binding proteins except transferrin were significantly increased in RA SF as compared with OA SF. Similarly, free iron was also significantly higher in RA SF than in OA SF, whereas the ferritin saturation index, transferrin saturation index and bound iron were more significantly decreased in RA SF than in OA SF. These results suggest that RA SF contains sufficient micromolar amounts of free iron to allow hydroxyl radical formation. Also the capacity of iron-binding proteins to bind free iron is inadequate in the presence of a large amount of iron-binding proteins which are present in RA SF.

Antigen-Antibody Complex↗
PubMed1996

Increased population of high fluorescence 1F7 (CD26) antigen on T cells in synovial fluid of patients with rheumatoid arthritis.

OBJECTIVE: To investigate the activation of T cells in peripheral blood (PB) and synovial fluid (SF) of patients with rheumatoid arthritis (RA). METHODS: The expression of CD26 (Ta1 and 1F7) antigen on T cells was analyzed in 7 women with RA and 7 healthy control subjects by immunofluorescence. RESULTS: The percentage of CD3+ CD26+ cells was significantly higher in PB of patients with RA compared with healthy subjects. The IF7+ cell population was divided into high (1F7+high cells) and low fluorescence populations (1F7+low cells), based on 1F7 antigen density. The percentage of 1F7+high cells in SF of RA was markedly increased compared with PB of patients and healthy subjects. However, RA SF contained lower percentages of whole 1F7+ cells compared with PB. CONCLUSION: Our results indicate that SF of patients with RA contains activated T cells, and suggest that T cells with high levels of CD26 antigen may preferentially migrate into the rheumatoid synovium to induce inflammation and tissue destruction.

Adult↗
PubMed1982

Functional characteristics of synovial fluid and blood mononuclear cells in rheumatoid arthritis and traumatic synovitis.

The activity of blood mononuclear cells (BMC) and synovial fluid mononuclear cells (SMC) from patients with rheumatoid arthritis (RA) and traumatic synovitis (TS) was assessed by means of [14C]thymidine incorporation and production of leukocyte migration inhibitory factor (LIF). When compared with normal controls, spontaneous LIF production by BMC was found in 5 of 9 TS patients, whereas spontaneous LIF production by rheumatoid arthritis BMC and by SMC from both patient groups was infrequently seen. ConA-induced LIF production by BMC and SMC from both patient groups did not differ significantly from that of normal controls. Thymidine incorporation by unstimulated SMC and BMC was low in both patient groups. After stimulation with polyclonal activators, SMC showed significantly reduced proliferation in comparison with BMC, but the responses to microbial antigens were equal to or higher than those of BMC. The proliferative responses of stimulated SMC from TS patients were higher than the responses displayed by stimulated SMC from RA patients.

Adult↗
PubMed1989

Evidence for activation of platelets in the synovial fluid from patients with rheumatoid arthritis.

Platelets were isolated by gel filtration from paired samples of peripheral blood and synovial fluid (SF) aspirated from inflamed knee joints from 20 adult patients with rheumatoid arthritis (RA) as well as from peripheral blood obtained from 20 healthy subjects. The platelets from the three different sources were investigated for quantitative differences in the number of two distinct types of intracellular storage organelles using immunofluorescence staining for platelet factor 4 (PF4) and labelling with the fluorescent substance mepacrine (MC). The number of PF4-stained organelles per cell was the same in the peripheral normal and RA platelets. This number was distinctly lower in the SF platelets. The peripheral and SF platelets from the RA patients had the same number of MC-labelled organelles. This number was distinctly lower than in the normal cells. The results suggest that the peripheral RA platelets had been activated to liberate serotonin and other substances from one type of organelles, and that the SF platelets had been activated to an additional liberation of PF4 from another such type. Liberated PF4, serotonin, and other substances from SF platelets may, in several ways, contribute to the inflammatory responses of RA.

Adult↗
PubMed1995

Detection and investigation of the molecular nature of low-molecular-mass copper ions in isolated rheumatoid knee-joint synovial fluid.

Low-molecular-mass copper(II) species have been detected and quantified in ultrafiltrates (n = 7) of rheumatoid synovial fluid (SF) by a highly-sensitive HPLC-based assay system with the ability to determine Cu(II) concentrations of < 10(-7) mol.dm-3. High field 1H NMR spectroscopy demonstrated that addition of Cu(II)(aq.) to isolated samples of RA SF ultrafiltrates resulted in complexation by histidine > alanine > formate > threonine > lactate > tyrosine > phenylalanine, their effectiveness in this context being in the given order. CD spectra of Cu(II)-treated samples of intact SF exhibited absorption bands typical of copper(II)-albumin complexes, in addition to a band attributable to a low-molecular-mass histidinate complex (lambda min 610 nm). Since both albumin and histidine are potent radical scavengers, these results indicate that any .OH radical generated from bound copper ions will be 'site-specifically' scavenged. Hence, low-molecular-mass copper complexes with the ability to promote the generation of .OH radical which can then escape from the metal ion co-ordination sphere (and in turn, cause damage to critical biomolecules) appear to be absent from inflammatory SF.

Adult↗
PubMed1988

Serum and synovial fluid levels of serum amyloid A protein and C-reactive protein in inflammatory and noninflammatory arthritis.

Serum and synovial fluid (SF) levels of serum amyloid A (SAA) and C-reactive protein (CRP) were measured in 46 cases of various inflammatory arthritis (Group 1), and in 40 cases of noninflammatory arthritis: 18 cases of osteoarthritis (Group 2) and 22 cases of traumatic arthritis (Group 3). Serum and SF SAA levels were markedly elevated in Group 1: 126.4 micrograms/ml +/- 19.2 SEM and 46.4 micrograms/ml +/- 10.5 SEM, respectively; moderately elevated in Group 2: 10.1 micrograms/ml +/- 2.9 SEM, 4.0 micrograms/ml +/- 1.1 SEM and moderately elevated in Group 3: 10.4 micrograms/ml +/- 1.2 SEM, 4.0 micrograms/ml +/- 1.2 SEM, respectively. Serum/SF SAA ratios were similar in all 3 groups and ranged between 2.52-2.72. In comparison to SAA, the increment of serum and SF CRP above normal levels was moderate. A positive strong correlation was found between serum SAA and serum CRP: r = 0.64 (p less than 0.001) and between SF SAA and SF CRP: r = 0.59 (p less than 0.0001). SF SAA did not correlate with the number of SF white blood cells but did correlate with the percent of SF polymorphonuclear cells: r = 0.23 (p less than 0.05).

Arthritis↗
PubMed1997

IgG2b inducing factor from rheumatoid arthritis synovial fluid synergizes with transforming growth factor-beta in promoting IgG2b antibody production in mouse B lymphocytes.

Rheumatoid arthritis synovial fluid (RA-SF) contains a distinct biological activity that selectively induces IgG2b production in LPS-activated murine B blasts. This IgG2b inducing factor (IgG2bIF) acts directly on purified LPS-activated B blasts from normal and Bruton's tyrosine kinase-defective CBA/N mice. In order to test the possibility that TGF-beta and IgG2bIF in RA-SF act in concert to induce IgG2b production, anti-TGF-beta monoclonal antibodies and RA-SF were added to LPS-activated CBA B blasts, which led to a marked reduction of IgG2b-producing cells. This result indicates that TGF-beta and IgG2bIF in RA-SF synergize in the induction of IgG2b production. TGF-beta antibodies do not inhibit IgG2b production in CBA/N B blasts, further substantiating our earlier notion that CBA/N B blasts have a higher endogenous production of TGF-beta after LPS stimulation, which might be responsible for the aberrant reactivity in btk deficient CBA/N mice. RA-SF is able to reconstitute the deficient IgG1 response in LPS- and IL-4-stimulated CBA/N B blasts. Addition of antibodies against TGF-beta had only a marginal effect on the IgG1 response, indicating that the reconstitution is mediated by another factor(s), which is present in RA-SF.

Animals↗
PubMed2006

Rheumatoid synovial fluid T cells are sensitive to APO2L/TRAIL.

The infiltration and accumulation of T cells in the rheumatoid arthritis (RA) synovial fluid (SF) are hallmarks of disease. We aimed to assess the functional relevance of FasL and of APO2L/TRAIL in the persistence of T cells in the rheumatoid SF. We have analyzed the expression of the activation markers HLA-DR and CD69 and also of the death receptor Fas/CD95 and death ligands FasL or APO2L/TRAIL in CD3+ lymphocytes from SF of 62 RA patients, together with their sensitivity to anti-Fas mAb or to rAPO2L/TRAIL, using as controls T lymphocytes present in SF of 20 patients with traumatic arthritis. T lymphocytes infiltrated in SF of RA patients have a chronically activated phenotype, but they are resistant to Fas-induced toxicity. However, they are more susceptible to rAPO2L/TRAIL than T cells in the SF of traumatic arthritis patients. In addition, we found very low amounts of bioactive FasL and APO2L/TRAIL associated with exosomes in SF from RA patients as compared with SF from traumatic arthritis patients. The observation on the sensitivity of RA SF T cells to rAPO2L could have therapeutic implications because bioactive APO2L/TRAIL could be beneficial as a RA treatment.

Antigens, CD↗