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Relations between absolute number of CD4 + CD29+ memory cells and levels of interleukin-6, rheumatoid factors, and acute phase proteins in rheumatoid synovial fluid.

Synovial fluid samples from 21 rheumatoid arthritis patients were analyzed for lymphocyte subsets using flow cytometry and antibodies to the lymphocyte surface antigens CD3, CD4, CD8, CD16, CD19, CD29, and CD45RA. Synovial fluid levels of interleukin-6, rheumatoid factors and acute phase proteins were also measured. Depletion of CD45RA+ cells and predominance of CD29+ cells were found. Interleukin-6 levels were markedly elevated (1155 pg/ml; range, 24-3875 pg/ml), as were levels of IgM, IgG and IgA rheumatoid factors and of acute phase proteins. CD4 + CD29+ counts were significantly correlated with interleukin-6 levels. Interleukin-6 levels were significantly correlated with levels of all three rheumatoid factor classes but not with levels of acute phase proteins. Significant correlations were also found between CD19+ B counts and levels of all three rheumatoid factor classes. These data suggest that synovial fluid CD4 + CD29+ cells may be involved in the immune dysregulation characteristic of rheumatoid arthritis. The correlation between CD4 + CD29+ counts and interleukin-6 levels is consistent with the recent hypothesis that, together with monocytes, CD4 + CD29+ cells are an important source of the elevated levels of interleukin-6 seen in rheumatoid synovial fluid.

Acute-Phase Proteins↗

Arthritis diagnosis based upon the near-infrared spectrum of synovial fluid.

Synovial fluid aspirates have been characterized by measuring their visible/near-infrared spectra (400-2500 nm). The hypothesis tested in this study is that the spectra contain sufficient information to serve as an aid in the diagnosis and/or staging of arthritic disorders. The concentrations of all major constituents are carried implicitly in the spectra, and in this sense this approach is similar in spirit to conventional synovial fluid analysis. The distinguishing feature of this method is that we have not converted the raw data (spectra) explicitly to analytical information. Rather, we have used automated pattern recognition methods to identify significant characteristics of the spectra themselves. A total of 109 spectra were measured and split into three classes according to the disease (osteoarthritis, rheumatoid arthritis, or spondyloarthropathy) affecting the patient from whom the synovial fluid sample was taken. An automated classification method was then trained by correlating features derived from these spectra to the clinical diagnoses. The robustness of the classification was validated using the leave-one-out cross-validation method, i.e., by training on all but one of the spectra and using the resulting model to predict the classification for the spectrum that is left out. The result derived by following this procedure for each of the spectra was that 105 of the 109 predicted classifications correctly matched the clinical diagnosis. These results suggest that the near-infrared spectrum of synovial fluid is sufficient to allow diagnosis of the disease affecting the joint from which the aspirate is drawn.

Arthritis, Rheumatoid↗

Phenotypes and spontaneous cell cytotoxicity of mononuclear cells from patients with seronegative spondyloarthropathies: ankylosing spondylitis, psoriatic arthropathy and pauciarticular juvenile chronic arthritis--analysis of mononuclear cells from peripheral blood, synovial fluid and synovial membranes.

T-cell subpopulations and natural killer (NK) cells from peripheral blood, synovial fluid and synovial membranes from patients with seronegative spondyloarthropathies were investigated. Thirty-four patients with ankylosing spondylitis, sixteen patients with psoriatic arthropathy and six patients with pauciarticular juvenile chronic arthritis were studied. All the patient groups had normal proportions of T4+ and T8+ cells as well as normal T4/T8 ratios in peripheral blood. In the synovial fluids the T4/T8 ratios were reduced in ankylosing spondylitis and psoriatic arthropathy (p less than 0.05). Although both the T4 and T8 subpopulations were reduced, the T4/T8 ratios in the synovial membranes of patients with these two disorders tended to be within the normal range of that of peripheral blood. Increased numbers of T-cells in the synovial fluid from patients with ankylosing spondylitis expressed class II MHC antigens. The natural killer cell activity was normal in peripheral blood and synovial fluids of patients with ankylosing spondylitis and psoriatic arthropathy while it tended to be reduced, although not significantly, in pauciarticular juvenile chronic arthritis. Synovial membranes were almost devoid of NK cell activity. The number of Leu 7+ cells were reduced in synovial fluid of patients with psoriatic arthropathy (p less than 0.04), but not as significantly as in the two other patient groups.

Adult↗

T cytotoxic and helper cells are markedly increased, and T suppressor and inducer cells are markedly decreased, in rheumatoid synovial fluids.

Synovial fluid lymphocytes and paired peripheral blood lymphocytes from 28 patients with rheumatoid arthritis were analyzed by 2-dimensional flow cytometry. We found that the composition of the lymphocyte population is different in synovial fluid compared with that in peripheral blood, in that there were increased proportions of Ia+ Leu-2a+/Leu-15- (T cytotoxic) cells and Leu-3a+/Leu-8- (T helper) cells, together with marked decreases in Leu-2a+/Leu-15+ (T suppressor) cells and Leu-3a+/Leu-8+ (T inducer) cells. These findings suggest that the unique composition of synovial fluid lymphocytes might be the result of T cell activation by some mechanism that is not known.

Adult↗

Characterization of antibody-dependent cell-mediated cytotoxicity in rheumatoid synovial fluid.

Synovial fluids of 15 rheumatoid arthritis patients, 11 juvenile rheumatoid arthritis patients, and 6 patients with other chronic inflammatory joint diseases were studied. The isolated mononuclear cells, mainly monocyte/macrophages and lymphocytes, were preincubated overnight and then cultured with antibody-sensitized 51Cr-labelled chicken erythrocytes. There was no significant difference in the cytotoxicity between mononuclear cells from synovial fluid and peripheral blood of the patients. Depletion of adherent cells of the mononuclear cell suspensions by nylon wool column fractionation or by incubation on plastic dishes and depletion of cells which had phagocytosed colloidal iron had no substantial effect on the cytotoxicity whereas depletion of cells forming rosettes with human erythrocytes sensitized with human IgG (EA-RFC) practically abolished the activity. This shows that in rheumatoid synovial fluids the net effect of monocyte/macrophages in this in vitro antibody-dependent cytotoxic assay is small. Thus, the main effector cells in these mononuclear cell suspensions are lymphocytes which probably exhibit surface-bound Fc-receptors.

Antibody-Dependent Cell Cytotoxicity↗

Antirheumatic drug concentrations in human synovial fluid and synovial tissue. Observations on extravascular pharmacokinetics.

Antirheumatic drug concentrations have been measured in human synovial fluid and synovial tissue, and provide insights on: (1) extravascular pharmacokinetics; (2) articular pathophysiology; and (3) the factors which modify drug levels in inflamed tissues. Concentrations of free drug in synovial fluid and plasma are the same in all conditions except rheumatoid and infectious arthritis, where the most severely afflicted joints may contain lower synovial fluid drug concentrations. This finding may be relevant to the chronicity and intractability of chronic arthritis. After single-dose therapy and a characteristic 'equilibration time', higher concentrations are found in synovial fluid than in plasma - a phenomenon which results from relative drug sequestration across the trans-synovial diffusion barrier away from the organs of elimination. Studies of oral, parenteral, topical and intra-articular antirheumatic drug therapy are reviewed, and recommendations are made for the conduct of future studies.

Anti-Inflammatory Agents↗

[Differential diagnostic value of nephelometrically determined protein fractions in the synovial fluid].

Synovial fluids of patients suffering from rheumatoid arthritis and osteoarthritis with effusions of the knees were examined. Different parameters were evaluated out of the synovial fluid (immunglobulins, Complement-1Q,-3,-4, haptoglobins, alpha-1-anti-trypsin, alpha-2-macroglobulin, transferrin, ceruloplasmin, rheumatoid factors, total count of cells, and ragocytes) and out of the plasma (blood sedimentation rate). The proteins were analysed by a nephelometricturbidimetric automatic centrifugal analyser. All parameters have been tested by valuable statistical methods and correlated to each other. The results worked out proved the reliability of the used test kits and apparative systems. Correlations within groups of parameters according to their formations (intra-and/or extraarticular) could not have been worked out in a way as it may be supposed. In contrast some parameters themselves are statistically different comparing rheumatoid arthritis and osteoarthritis. In general the results are on a higher level in the rheumatoid arthritis group. Using all parameters mentioned above the statistical differential diagnostic level is based on about 94%. If only blood sedimentation rate, total cell count and ragocytes are evaluated the level is based on 68%.

Acute-Phase Proteins↗

Use of BACTEC 9240 blood culture system for detection of Brucella melitensis in synovial fluid.

Synovial fluid specimens aspirated from patients with arthritis were inoculated into an aerobic Peds Plus blood culture bottle and monitored by the BACTEC 9240 instrument for 4 weeks. A total of 1,072 synovial fluid cultures were processed, and 15 (0.14%) were positive for Brucella melitensis. A single culture, harboring 1.3 CFU of viable organisms per ml, was missed by the instrument. All 14 positive BACTEC cultures were detected within 3 to 7 days.

Adolescent↗

Growth factors with heparin binding affinity in human synovial fluid.

Synovial effusions were obtained from the knees of 15 subjects with joint trauma, menisceal or ligamentous injury, or osteoarthritis. Heparin-Sepharose affinity chromatography of these synovial fluids revealed, in general, three major peaks of mitogenic activity as measured by incorporation of 3H-thymidine into 3T3 cells. Gradient elution patterns showed activities at 0.5M NaCl, which is characteristic of platelet derived growth factor, and at 1.1M NaCl and 1.6M NaCl, indicative of acidic and basic fibroblast growth factors, respectively. The identities of these mitogenic fractions were confirmed by specific immunologic and receptor-binding assays. The presence of platelet derived, acidic and basic fibroblast growth factors in the synovial fluid may contribute to wound healing in the arthritic joint.

Animals↗

Comparative analysis of CD45RA- and CD45RO-positive CD4+T cells in peripheral blood, synovial fluid, and synovial tissue in patients with rheumatoid arthritis and osteoarthritis.

To determine whether the predominant infiltration with memory CD4+T cells in joints is specific to the local immune and inflammatory response in rheumatoid arthritis (RA), the proportions of CD45RA+ or CD45RO+ cells in the CD4+T cell populations in three different compartments (i.e., peripheral blood, synovial fluid, and synovial tissue) from patients with RA and osteoarthritis (OA) were compared by two-color flow-cytometric analysis. In the CD4+T cell population of peripheral blood, the number of CD45RO+ cells was relatively higher than CD45RA+ cells in both RA and OA patients, but their percentages did not differ from those found in healthy individuals. However, the great majority of CD4+T cells present in synovial fluid and synovial tissue were CD45RO-positive and CD45RA-negative in both patient groups; although CD4+T cells infiltrating both the disease compartments were markedly greater in RA joints, their mean percentages of CD45RO+ cells were not significantly different from those in OA joints. These data indicate that an accumulation of CD45RO+ memory CD4+T cells is a generalized phenomenon during local inflammatory responses in both RA and OA joints, and may be due mainly to the propensity of these cells to preferentially transmigrate into the inflamed joint via adhesion molecules as compared with CD45RA+ naive CD4+T cells.

Aged↗

Effects of pulse methylprednisolone on inflammatory mediators in peripheral blood, synovial fluid, and synovial membrane in rheumatoid arthritis.

OBJECTIVE: To establish whether the clinical efficacy of pulse methylprednisolone (MP; 1,000 mg intravenously) is related to the modulation of proinflammatory cytokines within the peripheral blood, synovial membrane, or synovial fluid compartments. METHODS: Eighteen patients with active rheumatoid arthritis (RA) were studied. Peripheral blood (11 patients) and knee synovial fluid (9 patients, 10 knees) were obtained before and at 4 and 24 hours after MP therapy. Interleukin-1beta (IL-1beta), IL-8, and tumor necrosis factor alpha (TNFalpha) were measured by enzyme-linked immunosorbent assay and biologic assays; prostaglandin E2 (PGE2) was measured by competitive radioimmunoassay. In 10 patients, arthroscopically directed synovial biopsies were obtained before and at 24 hours after treatment, at disease relapse (4 patients), and after retreatment (1 patient). Membranes were stained by immunohistochemical techniques with monoclonal antibodies against TNFalpha, IL-8, IL-1beta, and the IL-1 receptor antagonist protein (IL-1Ra). RESULTS: MP therapy was associated with a rapid (within 24 hours) and substantial decrease in the expression of TNFalpha in the lining and sublining regions of the synovial membrane, as well as substantial decreases in the levels of TNFalpha in serum and synovial fluid. There was also reduced IL-8 expression in the synovial lining, as well as reduced synovial fluid IL-8 levels. No effect on synovial membrane IL-1beta and IL-1Ra or synovial fluid IL-1beta and PGE2 was found. CONCLUSION: MP therapy rapidly reduces IL-8 and TNFalpha levels in the synovial compartment, with cytokine changes in the serum and synovial fluid reflecting the changes in the synovial membrane. Alterations in TNFalpha expression in the synovial membrane correlated with clinical response to, and subsequent relapse after, MP therapy.

Aged↗

Fibronectin in rheumatoid and non-rheumatoid arthritic synovial fluids and in synovial fluid cryoproteins.

Concentrations of fibronectin, immunoglobulins G, M, and A, and C3 and C4 components of complement, and other plasma proteins were determined in synovial fluids from patients with rheumatoid arthritis (RA) and other diseases (non-RA). Fibronectin concentrations were two to three times greater in all synovial fluids than in plasma, and RA synovial fluids had a significantly higher mean concentration than non-RA fluids (883 microgram per ml vs. 588 microgram per ml, respectively, p less than 0.01). The mean concentrations of other synovial fluid constituents were less than their mean plasma concentrations. These results suggest that unlike other plasma constituents, either plasma fibronectin is concentrated in synovial fluids or that a substantial portion of synovial fluid fibronectin may be derived from synovial tissue cells. Both the C3 and C4 complement components were present in lower concentrations in RA than in non-RA synovial fluids. The C3 contents showed a statistically significant negative correlation with the fibronectin contents. Fibronectin was also found in all synovial fluid cryoprotein fractions tested, although its content varied greatly as a percent of the total cryoprotein protein (0.01 to 43 percent). The data show that fibronectin is a consistent constituent of synovial fluid cryoproteins in agreement with our previously reported finding that fibronectin is found in all serum cryoglobulin fraction tested.

Arthritis↗

[The distribution of benorylate in plasma, synovial fluid and synovial tissue in rheumatoid arthritis].

The authors report on the drug benorylate which is available in the Federal Republic under the name of Benortan (4-acetamidophenyl-2-acetoxybenzoat). Numerous examinations have shown that it is well tolerated by mouth and that it has beneficial clinical effects with few side effects. Benorylate is a neutral, fat-soluble, water-insoluble substance which upon absorption is almost completely hydrolyzed into salicylate and paracetamol. 6 patients with classical seropositive rheumatoid arthritis and with a highly active synovialitis of one or both knee joints not previously treated received 4 g of a 40% benorylate suspension orally twice daily over a period of 9-14 days. On different days at exactly determined times of drug administration blood, and on the day of synovectomy synovial fluid and synovial tissue, were taken and frozen to - 70 degrees C and subsequently examined as to the content of salicylate, paracetamol, and unchanged benorylate. The plasma levels of salicylate and paracetamol were generally distinctly higher than the concentrations of these metabolites in the synovial fluid. Benorylate which is practically not detectable in blood is found in the synovial tissue and is detectable in greatest quantities in the most inflamed synovial villi. Benorylate can probably penetrate into the synovial membrane like its metabolites salicylate and paracetamol; it remains, however, to be examined whether the metabolites are distributed differently in different synovial areas (active inflamed and unattacked synovial tissue, respectively) in the same way as benorylate per se.

Acetaminophen↗

Synovial tissue and synovial fluid.

Synovial tissue is a highly specialized tissue that keeps the articular joint well lubricated, and at the same time provides nutrients to the articular surface. A joint needs a small amount of synovial fluid to work. Although the tissue structure is simple, its function is highly sophisticated. Because synovial tissue is the target of most autoimmune diseases, joints need to be understood in their entirety to create new treatments for such immunological disorders.

Adult↗

Clonality of T lymphocytes expanded with IL-2 from rheumatoid arthritis peripheral blood, synovial fluid and synovial membrane.

The association of rheumatoid arthritis (RA) with particular MHC class II genes suggests that autoantigen-specific T cell clones present in joints could be central to the pathogenesis of the disease. Previous investigations on the clonal diversity of T cells infiltrating the rheumatoid synovial membrane have yielded conflicting results. With the use of Southern blot analysis, we investigated the clonality of rheumatoid T cell lines expanded from peripheral blood, synovial fluid and synovial tissue. From peripheral blood lymphocyte (PBL) of RA patients and healthy normal controls, we also checked the consequences of two different culture conditions on the clonality of these cell lines. From control PBL, we found that in vitro non-specific expansion of non-clonal T cell populations does not create artefactual clonal selection. However, growing T cells in vitro with IL-2 seems to be able to lead to preferential expansion of cells bearing IL-2 receptor (IL-2R). We identified such in vivo activated IL-2-sensitive T cell clones frequently in RA synovial tissue (8/13) and more rarely in synovial fluid and peripheral blood (3/12). One patient presents the same T cell receptor gene rearrangements in synovial membrane of two affected joints. In RA synovial tissue, the frequency of these IL-2-responsive T cells is most prevalent among actively inflamed membranes removed early in the disease process. The role and the relevance to the disease of these IL-2-responsive T cells remain to be elucidated.

Adult↗

Natural killer (NK) cell activity of peripheral blood, synovial fluid, and synovial tissue lymphocytes from patients with rheumatoid arthritis and juvenile rheumatoid arthritis.

Natural killer (NK) cell activity was investigated in peripheral blood, synovial fluid, and synovial tissue lymphocytes from patients with rheumatoid arthritis (RA) and juvenile rheumatoid arthritis (JRA). Unfractionated lymphocytes, T lymphocytes, and non-T lymphocytes from the 3 compartments of JRA patients had reduced activity compared with that of normal peripheral blood lymphocytes (with p values usually between 0.05 and 0.1). Unfractionated synovial tissue lymphocytes of RA patients also showed reduced cytotoxicity (0.05 less than p less than 0.1), whereas peripheral blood lymphocytes exerted normal NK cell activity. The NK activity was exerted by cells both with and without Fc gamma receptors. The highest cytotoxicity was observed in Fc gamma receptor-positive cells, both in peripheral blood and synovial fluid, since more than 70% reduction in NK activity was found after depletion of Fc gamma receptor-positive cells. No evidence of lymphocytotoxic antibodies or other factors with influence on NK cells was observed in the patients' sera.

Adolescent↗

[A study of opioid peptides in synovial fluid and synovial tissue in patients with rheumatoid arthritis].

Authors have often experienced that psychological stress influences rheumatoid arthritis (RA). In addition, recent reports show a modulatory role for neuropeptide such as substance P in arthritis. These findings prompted us to study endogenous opioid peptides in RA, which are found mainly in the brain and have an effect on the central nervous system. We examined methionine-enkephalin (Met-enk), leucine-enkephalin (Leu-enk) and beta-endorphin (beta-end) in opioid peptides. We measured these peptides in plasma and synovial fluid samples obtained from 28 knees of 24 RA patients and the quantity in the synovial tissue of 13 knees. We also measured plasma and synovial fluid samples from patients with osteoarthritis of the knee and plasma samples from healthy candidates. Leu-enk and beta-end levels in synovial fluid were significantly higher than plasma levels only in RA. Larger quantity of Leu-end and beta-end were contained apparently in the synovial tissue than Met-enk. The synovial tissue with proliferative change tends to contain larger quantity of opioid peptides. These results indicate that the synovial tissue produces or secretes Leu-enk and beta-end and that opioid peptides are related to the degree of inflammation in RA.

Adult↗

Cytokines and the chronic inflammation of rheumatic disease. I. The presence of interleukin-1 in synovial fluids.

Synovial fluids from patients with rheumatoid arthritis (13 cases), ankylosing spondylitis (six), psoriatic arthritis (two), osteoarthritis (three) and rubella arthritis (four) contain interleukin-1 (Il-1) activity as measured in the CH3/He mouse thymocyte assay. That this activity is Il-1 was shown by: (i) the time course of thymocyte stimulation; (ii) failure of PHA blast cells to absorb out the activity; (iii) molecular weight of 15,000-20,000 daltons, and (iv) ability to stimulate prostaglandin E2 production from synovial cells. These results indicate that IL-1 is an important mediator in the inflammatory pathway of different types of joint disease.

Animals↗