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Complement-derived leukotactic factors in inflammatory synovial fluids of humans.

A large per cent of rheumatoid synovial fluids contain chemotactic activity for rabbit granulocytes (neutrophilic). The chemotactic activity is, in large part, related to the fifth (C5) and sixth (C6) components of human complement; a combination of physical-chemical techniques indicates the activity to be attributable to C567 and C5a, a cleavage product of C5. Many rheumatoid synovial fluids contain a C5-cleaving enzyme which, on the basis of substrate specificity and susceptibility to inhibitors, is very similar to an enzyme extractable from lysosomal granules of human and rabbit granulocytes. Inflammatory nonrheumatoid synovial fluids contain chemotactic activity that is related to cleavage products (C3a) of the third component of human complement (C3). Also found in these fluids is a C3-cleaving enzyme capable of producing C3a. Of the other synovial fluids examined, lupus fluids were remarkable by their total lack of chemotactic activity. These findings record for the first time the presence of complement-derived chemotactic factors in pathological human fluids.

Antigen-Antibody Reactions↗

[Antibodies to rubella virus in the blood sera and in the synovial fluid of children with rheumatoid arthritis].

Sera and synovial fluids from children with rheumatoid arthritis (RA) were examined for antibody to some viral and mycoplasmal antigens. Blood sera from the patients were found to have a selective excess of antibody to rubella virus as compared with the age norm. Antibody of this specificity was found in synovial fluids in titers significantly higher than those in the blood. Titers of other antibodies in the synovial fluids were, as a rule, slightly lower than in sera. In the time course of intercurrent respiratory diseases, despite the lack of rubella introduction into the wards, children with RA showed variations in the levels of antibody to rubella virus much more frequently than to other infectious agents. The foregoing data are discussed from the point of view of the etiological association of juvenile RA with rubella virus persistence. The antigen of the latter was found in snyovial membrane cells of 4 children by means of immunofluorescence procedure.

Adolescent↗

Rheumatoid factor has increased reactivity with IgG from synovial fluids of patients with rheumatoid arthritis and osteoarthritis.

Synovial fluid IgG may be altered in rheumatoid arthritis (RA) and promote the formation of immune complexes with rheumatoid factor. To investigate this possibility, monomeric IgG was prepared from synovial fluids from a range of arthritides for use as the antigen in a rheumatoid factor microplate radioimmunoassay. In comparisons with normal serum IgG antigen, increased rheumatoid factor binding was shown to IgG antigens prepared from synovial fluids from patients with RA and osteoarthritis (OA). Increased binding was also shown to RA sera IgG, but not to OA sera IgG. This increased binding was not due to increased IgG antigen binding to the plate or to IgG rheumatoid factor in the antigen preparations. It was considered that cause was a structural alteration of the IgG as a result of inflammation within the rheumatoid and OA joint.

Antigens↗

The application of atomic force microscopy for the detection of microcrystals in synovial fluid from patients with recurrent synovitis.

Synovial fluid from 33 patients with inflammatory arthritis was examined with a polarized light microscope (PLM) and an atomic force microscope (AFM). Two samples were imaged with a transmission electron microscope (TEM) to determine calcium/phosphate ratios and identify microcrystals of calcium pyrophosphate dihydrate and octacalcium phosphate. Additional correlative x-ray diffraction studies were performed on several samples including purified hydroxyapatite and sodium chloride crystals. Monosodium urate, calcium pyrophosphate dihydrate, hydroxyapatite, octacalcium phosphate, and cholesterol crystals were identified with AFM. AFM images of these microcrystals revealed detailed surface topology, including lattice parameters and structural irregularities at the crystals' surface. These features were consistent with those obtained by TEM and x-ray diffraction studies. In addition, AFM images revealed that some specimens contained microcrystals that were undetected by PLM and/or TEM. These results suggest that AFM may provide a simple yet powerful technique for the detection of microcrystals in synovial fluid taken from patients with crystal-induced arthritis.

Arthritis↗

Interleukin-6 and interleukin-8 levels in serum and synovial fluid of patients with osteoarthritis.

Concentrations of interleukin (IL)-6 and IL-8 in serum and synovial fluid obtained from patients with osteoarthritis (OA) of the knee were determined by the chemiluminescence-ELISA (CL-ELISA) method, the sensitivity of which is 100-1,000 times greater than that of the conventional ELISA method. The results were compared with those obtained from patients with rheumatoid arthritis (RA) and from healthy subjects. The mean IL-6 and IL-8 levels in synovial fluid indicated higher concentrations in RA than in OA. The IL-6 and IL-8 levels in serum were significantly higher in RA and OA relative to controls. Among OA patients in whom remarkable improvement was noted in hydrarthrosis, the synovial fluid IL-6 and IL-8 levels at the initial examination were relatively higher, and were markedly decreased after treatment with sodium hyaluronate (NaHA). Among those in whom no improvement was noted in hydrarthrosis, the synovial fluid IL-6 and IL-8 levels at the time of initial examination were relatively lower, and hydrarthrosis was not significantly improved even after treatment with NaHA. In addition, there was a tendency for the synovial fluid IL-6 and IL-8 levels to decrease as HA levels increased. Evaluation of X-ray findings revealed that the IL-6 levels in synovial fluid at the initial examination in low-grade cases tended to be significantly higher than in high-grade cases. In low-grade cases, as determined by X-ray findings, there was a significant decrease in IL-6 levels in synovial fluid after treatment with NaHA.

Adjuvants, Immunologic↗

Synovial fluid and serum hydroxyproline fractions in rheumatoid arthritis.

In rheumatoid arthritis (RA) the synovial fluid lymphocyte count closely parallels the synovial fluid dialysable hydroxyproline, a marker of collagen resorption. This observation stresses the primordial role of lymphocytes in RA joint injury. The actual and eventually potential destruction of any single joint, as expressed by synovial fluid dialysable hydroxyproline levels, seems to reflect the general picture of disease activity as evaluated by the number of active joints (joint count), ESR and rheumatoid factor titres (latex fixation). Although urinary hydroxyproline levels are generally within the normal range in RA, they can be used as index of articular tissue destruction and as a parameter of overall disease activity when groups of patients are studied longitudinally in detail. The present study sheds no further light on the significance of synovial fluid and blood non-dialysable hydroxyproline levels.

Arthritis, Rheumatoid↗

Correlation between rheumatoid factor and IL-6 activity in synovial fluids from patients with rheumatoid arthritis.

Synovial fluids (SF) and sera (S) from patients with rheumatoid arthritis (RA) were examined for IgM, IgM-rheumatoid factor (IgM-RF), albumin and interleukin-6 (IL-6) activity. The quotient of SF/S IgM-RF was elevated compared with that of SF/S albumin in 7 patients with seropositive RA, although the quotient of SF/S IgM was lower than that of SF/S albumin. SF IL-6 activity was much higher than serum IL-6 activity in all the 7 RA patients. In synovial fluids from 22 seropositive RA patients, SF IL-6 activity was significantly correlated with the SF IgM-RF, IgG-RF and IgA- less than RF, but not with SF IgM, IgG or IgA. Moreover, SF IgM-RF as well as SF IL-6 activity was significantly correlated with the Westergren erythrocyte sedimentation rate (ESR) or the Lansbury articular index. These results indicate that IL-6 and RF might be produced within the rheumatoid joints as a result of abnormal immune system activation, which is associated with the disease activity of RA. Three of the 4 seronegative RA patients, however, showed high SF IL-6 without detectable levels of SF IgM-RF, indicating that IL-6 alone is not sufficient for IgM-RF production.

Adult↗

Clonal heterogeneity of synovial fluid T lymphocytes from patients with rheumatoid arthritis.

Although substantial evidence suggests that synovial T lymphocytes are critical in the pathogenesis of rheumatoid arthritis (RA), little is known regarding their antigenic specificities, antigen receptor gene rearrangements, and mechanisms of activation. To assess the extent of expansion of specific clones among RA synovial fluid T cells, Southern blot analyses of T-cell receptor (TCR) gene rearrangements were performed on 40 RA synovial fluid T-cell clones, as well as on both fresh and polyclonally activated T cells from RA synovial fluid, RA peripheral blood, and normal peripheral blood. Two of the clones had identical TCR rearrangement patterns, but the remainder were unique. The nonclonal RA T-cell samples showed the same pattern of TCR beta-chain rearrangement that was observed among normal peripheral blood T cells, indicating no dominant clonal T-cell population in these samples. It was noted that with sufficient exposure of autoradiograms of the Southern blots, discrete TCR gene rearrangements, representing in some cases common D beta J beta (D, diversity; J, joining) rearrangements, were evident in T cells from peripheral blood of normal individuals and patients with RA, as well as T cells from RA synovial fluid. Taken together, the findings indicate that only a minor degree of oligoclonality can be demonstrated among T lymphocytes from RA synovial fluid.

Arthritis, Rheumatoid↗

Gonadal and adrenal steroid hormones in plasma and synovial fluid of patients with rheumatoid arthritis.

OBJECTIVES: Gonadal and adrenal steroids were shown to affect multiple immune processes including inflammatory response. These effects were documented, specifically, through an influence on local productions of cytokines and the functions of synovial cells at the site of inflammatory processes. The aim of this study was to investigate the levels of selected hormones in synovial fluid of knee joints of patients with rheumatoid arthritis (RA) and with osteoarthrosis (OS, control group). METHODS: The concentrations of cortisol (CORT), 17-beta-estradiol (ES), dehydroepiandrosterone (DHEA), testosterone (TE), progesterone (PRG), and aldosterone (ALD) were determined in plasma and synovial fluid. RESULTS: Significant positive correlations between the levels in plasma and synovial fluids were observed in hormones ES, PRG, TE, DHEA and ALD. In most hormones, the levels in synovial fluids were similar as in plasma; however, the content of ALD was higher in synovial fluid as compared to plasma. Higher levels of ES (in females), DHEA (in males), and ALD were observed in plasma and synovial fluids of RA patients as compared to OS patients. After adjustment to age, no significant RA vs. OS difference was noted in ES, TE, DHEA, PRG, and CORT in plasma and synovial fluid. Age-adjusted ALD concentration tended to be higher in synovial fluid of RA patients as compared to OS patients. The ratio of ES/TE concentrations in synovial fluid was significantly higher in male RA patients compared to OS group. Also the ES/CS and ES/DHEA ratios in synovial fluid were elevated in RA patients in comparison to controls. CONCLUSIONS: These results demonstrated the prevalence of pro-inflammatory hormones in synovial fluid of RA patients, suggesting the possible role of these steroid hormones in inflammatory processes.

Adrenal Cortex Hormones↗

Synovial fluid and serum concentrations of aminoterminal propeptide of type III procollagen in healthy volunteers and patients with joint disease.

OBJECTIVES: To analyse synovial fluid and serum concentrations of the amino-propeptide of the type III procollagen (PIIINP) in normal individuals and patients with joint disease, and to explore the relationship between synovial fluid PIIINP concentrations and the rheumatological diagnosis, local inflammation, and joint disease. METHODS: A radioimmunoassay was used to measure the PIIINP concentrations in serum and knee joint synovial fluid from 16 healthy volunteers and patients with osteoarthritis (OA) (n = 40), rheumatoid arthritis (RA) (n = 30), and psoriatic arthritis (PsA) (n = 12). The PIIINP measurements were related to demographic data, synovial fluid leucocyte counts, and radiographic changes at the knee. RESULTS: Serum PIIINP concentrations were greater in each of the disease groups than in control subjects, but there were no differences between the disease groups. Synovial fluid concentrations of PIIINP were much greater than those in serum, indicating local production, and were significantly greater in RA than in other disease groups (p < 0.001). There was only a weak positive correlation between synovial fluid leucocyte counts, some radiographic changes, and synovial fluid PIIINP concentrations. CONCLUSIONS: These data suggest that synovial fluid PIIINP concentrations may reflect local synovial proliferative processes in joint disease, and that they could be of diagnostic and prognostic value in inflammatory arthropathies.

Adult↗

Use of the isolator 1.5 microbial tube for culture of synovial fluid from patients with septic arthritis.

Synovial fluid specimens obtained from patients with arthritis were plated onto solid media (conventional cultures) or inoculated into an Isolator 1.5 microbial tube (Isolator cultures), and the yield and time to detection of organisms were compared. Overall, 144 specimens obtained from 137 patients were processed, and 31 (21.5%) cultures obtained from 29 patients were positive by at least one method. Staphylococcus aureus was isolated from 12 patients, Streptococcus pneumoniae and Kingella kingae were isolated from 4 patients each, group G streptococci were isolated from 3 patients, Staphylococcus epidermidis and members of the family Enterobacteriaceae were isolated from 2 patients each, and Streptococcus mitis and Peptostreptococcus prevotii were isolated from 1 patient each. Overall, the causative organism was detected in 31 of 31 (100.0%) Isolator cultures and 24 of 31 (77.4%) conventional cultures (P < 0.02). Twenty-nine of 31 (93.5%) positive Isolator cultures and 20 of 24 (83.3%) conventional cultures were positive by the second day of incubation. Among the 24 cultures positive by both methods, higher numbers of CFU per milliliter were detected with the Isolator system in 13 cultures and with conventional cultures in 2 cultures (P < 0.002). Inoculation of synovial fluid into an Isolator 1.5 microbial tube improves the recovery of organisms causing septic arthritis.

Adult↗

Pharmacokinetics of the R(-) and S(+) enantiomers of ibuprofen in the serum and synovial fluid of arthritis patients.

Eight patients with arthritis and knee effusions received 13 doses of a single 800-mg ibuprofen tablet every 8 hours. Serum and synovial fluid samples were obtained after the first and last doses and assayed for the R(-) and S(+) enantiomers of ibuprofen by a stereospecific assay. Since only S(+)-ibuprofen inhibits cyclo-oxygenase, a description of the time course of this isomer in synovial fluid is needed for the development of suitable pharmacodynamic models. The isomers were significantly different with respect to peak concentrations and areas under the concentration-time curves (AUC) in synovial fluid levels. No significant accumulation of either isomer was observed in serum or synovial fluid levels between the first and the last doses. The steady-state concentration of both isomers fluctuated less in synovial fluid than in plasma, and the synovial fluid concentrations of the S(+) isomer were about twice that of the R(-) isomer. The mean synovial albumin concentration was about 60% of the serum albumin concentration, and the steady-state isomer AUC values in synovial fluid were significantly correlated with the corresponding serum values after the differences between the two fluids with respect to albumin concentration were corrected. The authors conclude that binding of the isomers to albumin and the serum-synovial fluid albumin ratio controls the steady-state distribution of the ibuprofen isomers into synovial fluid. The ramifications of these findings in the development of satisfactory concentration-response relationships are discussed.

Adult↗

[Sonographic findings for synovial fluid].

OBJECTIVE: The aim of this pictorial essay was to evaluate the sonographic features of synovial fluid in patients with arthritis. METHODS: Sixty-nine patients with active synovitis (rheumatoid arthritis, psoriatic arthritis, osteoarthritis, septic arthritis, crystal arthropathies, post-traumatic arthritis) were studied. Sonographic evaluation was performed with a AU-5 Harmonic, Esaote Biomedica (Genoa, Italy) equipped with a 10-14 MHz broadband linear transducer and a Diasus Dynamic Imaging Ltd.(Livingston, Scotland UK) equipped with a 8-16 MHz broadband linear transducer. RESULTS: Six main different sonographic patterns were detected: 1) Anechoic: increased amount of homogeneous anechoic synovial fluid (exudative synovitis). 2) Cloudy: echogenic structures (proteinaceous material). 3) Mixed: anechoic synovial fluid and proteinaceous material. 4) "Snow-storm" aspect: multiple mildly and heterogeneous echoic spots (9 out of 10 patients with acute gouty synovitis). 5) Dotted: multiple sparkling hyperechoic dots without posterior acoustic shadow (10 out of 12 patient with chondrocalcinosis). 6) Granular: irregular turbid aspect of the synovial fluid. It was present in 3 patient with septic arthritis. CONCLUSIONS: The results of this study indicate that high resolution ultrasonography is able to detect different features of synovial fluid. Further studies are needed to assess both sensitivity and specificity of ultrasonography in "in vivo" synovial fluid examination.

Arthritis, Infectious↗

Identification and functional importance of plasma kallikrein in the synovial fluids of patients with rheumatoid, psoriatic, and osteoarthritis.

OBJECTIVES: To determine and identify, unequivocally, if plasma kallikrein (PK) is present in the synovial fluid of patients with rheumatoid (RA), psoriatic (PA) and osteo (OA) arthritis, and to consider its functional importance in the inflamed joint. METHODS: Therapeutically aspirated synovial fluids (pooled and individual samples, n = 66) were obtained from patients with arthritis. In addition, serum (n = 14) was collected from RA patients, and saliva (n = 10) and urine (n = 10) from normal individuals. Enzymic (amidase) and immunoreactive activities of PK and its precursor, prokallikrein (PPK), were determined. The presence of PK was assessed by incubation with soya bean trypsin inhibitor (SBTI), and by adsorption with anti-PK antibody linked to Sepharose. An enzyme-linked immunosorbant assay (ELISA) for PK was developed for quantitative measurement of total PK in biological fluids. Enhancement of the PK dose-response by RA synovial fluid made it necessary to remove RF from synovial fluids before determination of PK by ELISA. RESULTS: Amidase activity was demonstrated in synovial fluid pools and shown to be inhibited completely by SBTI, and removed by prior treatment with anti-PK Sepharose. Total PK activity (PK + PPK) from individual synovial fluid specimens did not differ significantly between patients with RA (median activity 76 mU/g protein), PA (80 mU/g protein) or OA (60 mU/g protein). Similar results were obtained when active PK alone was measured. No correlation was found between active PK or total PK values and the severity score for individual joints. Most of the measured immunoreactivity was removed by adsorption with anti-PK antibody linked to Sepharose. CONCLUSION: The results support the hypothesis that plasma kallikrein is present in synovial fluid. The enzyme may be important in the pathogenesis of inflamed joints.

Adult↗

Correlation of some cytomorphological and ultrastructural modifications of synovial fluid in juvenile chronic arthritis.

Experiments have been performed on 25 synovial fluid samples from patients with juvenile chronic arthritis (mono- and polyarticular forms) and with hydroarthrosis, the latter considered as controls. By cytomorphologic studies, we determined the cellularity, ragocytosis and synoviocytogram of the synovial fluid cellular pellet and found out that the synovial fluid from cases of juvenile chronic arthritis is characterised by cytosis (11.270/mm3; 15.275/mm3), polynucleosis (67.3%, 72.2%) and ragocytosis (12.8%, 17.5%) whereas hydroarthrosis synovial fluid is characterised by lymphocytosis (47.8%). Ultrastructurally, ragocyte-like polymorphonuclear cells are characterised by: a) segmentation of the nucleus and preferential concentration of chromatime on the periphery of the nuclear membrane: b) frequent intracytoplasmic inclusions and phagolysosomes. Phagocyte-like mononuclear cells present numerous inclusions and phagolysosomes, certainly indicating an endocytic activity. Lymphocytes are characterised by a narrow cytoplasmic rim, presenting relatively few cellular organelles. They coexist with immunely activated lymphocytes rich in cytoplasm, mitochondria and endoplasmic reticle. Corroboration of cytomorphological and ultrastructural date enables us to explain the morphological modifications and emphasize their importance in juvenile chronic arthritis pathogenesis and diagnosis.

Arthritis↗

Characterization of the natural killer-like lymphocytes in rheumatoid synovial fluid.

IgG Fc- cytotoxic cells found in the synovial fluid of patients with rheumatoid arthritis have natural killer (NK)-like characteristics but can kill NK-resistant cell lines as well. The phenotype of these cells was defined by complement-mediated lysis with monoclonal antibodies. The synovial fluid killer cell activity was significantly reduced by treatment with complement and OKT11 and 4F2, but the cytotoxic T cells did not express the NK-related antigens OKM1 and Leu-7, nor the cytotoxic T lymphocyte-specific antigen, OKT8. These results demonstrate that the synovial fluid killer cells resemble the activated T cells generated in an autologous mixed leukocyte reaction or in the treatment of peripheral blood mononuclear cells with interleukin 2, and they are distinct from the conventional NK cells found in blood.

Adult↗

Serum and synovial fluid concentrations of keratan sulfate and hyaluronan in dogs with induced stifle joint osteoarthritis following cranial cruciate ligament transection.

OBJECTIVE: To examine longitudinal changes in serum and synovial fluid concentrations of keratan sulfate (KS) and hyaluronan (HA) after cranial cruciate ligament (CCL) transection in dogs. ANIMALS: 12 clinically normal adult mixed-breed dogs. PROCEDURE: Following CCL transection in the right stifle joint, KS and HA concentrations were determined in serum and neat (undiluted) synovial fluid prior to and 1, 2, 3, and 12 months after surgery. Postsurgical dilution of synovial fluid was corrected by use of urea as a passive marker. RESULTS: Synovial fluid KS and HA concentrations decreased at 1, 2, and 3 months after surgery in operated stifle joints, compared with baseline values. Synovial fluid KS concentration decreased in unoperated stifle joints at 1 month. A decrease in synovial fluid KS concentration was found in operated stifle joints, compared with unoperated stifle joints, at 2 and 3 months, and a decrease in synovial fluid HA concentrations was also found in operated stifle joints, compared with unoperated stifle joints, at 1, 2, and 3 months. Serum KS concentrations increased from baseline values at 3 months after surgery. Hyaluronan concentrations in operated stifle joints were lower than baseline values at 1, 2, and 3 months. Urea-adjusted synovial fluid concentrations revealed that dilution did not account for the decline in biomarker concentrations. CONCLUSIONS AND CLINICAL RELEVANCE: The initial decrease and subsequent increase in synovial fluid concentrations of HA and KS may be caused by an acute inflammatory response to surgical intervention that negatively affects cartilage metabolism or an increase in production of immature proteoglycans.

Animals↗

Impaired interleukin-8-dependent chemotaxis by synovial fluid polymorphonuclear leukocytes in rheumatoid arthritis.

The accumulation of polymorphonuclear leukocytes (PMN) in synovial fluid is a common feature of rheumatoid arthritis (RA). We studied the chemotactic response of PMN obtained from the synovial fluid and from the peripheral blood of patients with RA using a modified Boyden's method, in which interleukin-8 (IL-8) or N-formyl-methionyl-leucyl-phenylalanine (FMLP) was used as a chemotactic agent. The IL-8-induced response of peripheral blood PMN from 15 patients with RA did not differ from that of 15 healthy controls. A decreased chemotactic response to IL-8 was, however, observed in PMN from the synovial fluid of 12 patients with RA compared with peripheral blood cells of the same individual. This defective chemotactic ability of PMN was inversely correlated with the number of infiltrating cells in the synovial fluid. We also obtained similar results with FMLP. These results indicate that the chemotactic ability of PMN may be reduced after migrating to the synovial fluid.

Adult↗