interleukin-8 and Synovitis

Topics: Animals; Anti-Inflammatory Agents; Antibodies; Antibody Specificity; Arthritis, Rheumatoid; Autoantibodies; Blood Coagulation Factors; Chemokine CXCL5; Chemokines; Chemokines, CC; Chemokines, CXC; Chemotaxis, Leukocyte; Humans; Interleukin-8; Peptides; Receptors, Chemokine; Synovial Fluid; Synovitis

We evaluated the ability of Coll2-1, a type II collagen peptide, to activate pro-inflammatory pathways in synovial cells and to induce arthritis in Lewis rats.. Human synoviocytes and chondrocytes from knee OA patients were cultured for 24 h with/without Coll2-1 and/or purified immunoglobulin G (AS0619) binding specifically this peptide, and/or CLI-095, a TLR-4 signaling inhibitor and/or apocynin and diphenyleneiodonium, Reactive oxygen species (ROS) production inhibitors. The Interleukin (IL)-8 and Vascular Endothelium Growth Factor (VEGF) expression, the IL-8 production, the IκB-α and p65 phosphorylation and ROS were evaluated. Coll2-1 peptide, bovine type II collagen (CIA), streptococcal cell wall (SCW) or saline solution were injected into Lewis rats. The Coll2-1 peptide was injected subcutaneously (SC; 20-200μg/100μl/animal) or intra-articularly (IA; 0.5-5μg/50μl/animal) and compared to CIA injected in SC (200μg/100μl/animal) and SCW in IA (5μg/50μl/animal). The animals were injected on day 0 and monitored for 28 days. Histological lesions assessment was performed using an arthritis score.. Coll2-1 peptide significantly increased IL-8 gene expression and production by synoviocytes. AS0619 and CLI-095 significantly decreased IL-8 expression. Coll2-1 induced p65 and IκBα phosphorylation and oxidative stress inhibitors decreased it. In human chondrocytes culture, Coll2-1 significantly increased MMP-3 and VEGF gene expression. In Lewis rats, CIA, SCW or Coll2-1 injection triggered arthritis. Like CIA or SCW, Coll2-1 induced synovitis, loss of cartilage proteoglycans, cartilage structure lesion and subchondral bone remodeling.. Coll2-1 activates synoviocytes to produce IL-8 and induces arthritis in rat. These findings suggest that neutralizing Coll2-1 could be a therapeutic approach of arthritis.

Topics: Aged; Animals; Cells, Cultured; Collagen Type II; Disease Models, Animal; Female; Gene Expression Regulation; Humans; Interleukin-8; Male; Middle Aged; Oxidative Stress; Peptide Fragments; Rats; Rats, Inbred Lew; RNA; Synoviocytes; Synovitis

The objective was to study both ex vivo and in vitro secretion of pro-inflammatory cytokines in patients affected by Blau syndrome (BS) and carrying p.E383K mutation in the CARD15/NOD2 gene associated with the disease. For ex vivo studies, peripheral blood mononuclear cells (PBMCs), serum from three patients and healthy controls have been collected. PBMCs have been cultured in the presence or absence of inflammatory enhancers, such as lipopolysaccharide (LPS) and muramyl dipeptide (MDP). The levels of interleukin (IL)-1β, IL-6, IL-8, tumor necrosis factor (TNF)-α and interferon (IFN)-γ were assayed by either immunoassay or array-based system. For in vitro studies, different constructs were created cloning human wild-type and p.E383K-mutated NOD2 cDNA into the expression vector pCMV-Tag2c. HEK293 cell lines were stably transfected, cultured with or without MDP and IL-8 level was assayed in their surnatants. Statistical analysis in both studies was performed using non-parametric tests. Both ex vivo and in vitro studies have not identified a significant increase in secretion of the analyzed proinflammatory cytokines. p.E383K-mutated NOD2 transfected cells express low level of IL-8. The ex vivo basal level results from both serum and PBMCs surnatants present similar levels of IL-1β, IL-6, TNF-α and IFN-γ in patients and controls. The presence of the stimulant agents (LPS and MDP), either individual or paired, does not lead to significant increases in all cytokines concentrations in patients compared to controls. Taken together, the ex vivo and in vitro data suggest that there is not a primary mediation of IL-1β and other pro-inflammatory cytokines in BS patients carrying p.E383K.

Topics: Adult; Arthritis; Biomarkers; Case-Control Studies; Cytokines; Extremities; Fathers; Female; Humans; In Vitro Techniques; Interferon-gamma; Interleukin-1beta; Interleukin-6; Interleukin-8; Male; Middle Aged; Mutation; Nod2 Signaling Adaptor Protein; Nuclear Family; Pedigree; Sarcoidosis; Synovitis; Tumor Necrosis Factor-alpha; Uveitis

Despite the widespread use of magnetic resonance imaging (MRI) and Doppler ultrasound for the detection of rheumatoid arthritis (RA) disease activity, little is known regarding the association of imaging-detected activity and synovial pathology. The purpose of this study was to compare site-specific release of inflammatory mediators and evaluate the corresponding anatomical sites by examining colour Doppler ultrasound (CDUS) and MRI scans.. RA patients were evaluated on the basis of CDUS and 3-T MRI scans and subsequently underwent synovectomy using a needle arthroscopic procedure of the hand joints. The synovial tissue specimens were incubated for 72 hours, and spontaneous release of monocyte chemoattractant protein 1 (MCP-1), interleukin 6 (IL-6), macrophage inflammatory protein 1β (MIP-1β) and IL-8 was measured by performing multiplex immunoassays. Bone marrow oedema (BME), synovitis and erosion scores were estimated on the basis of the rheumatoid arthritis magnetic resonance imaging score (RAMRIS). Mixed models were used for the statistical analyses. Parsimony was achieved by omitting covariates with P > 0.1 from the statistical model.. Tissue samples from 58 synovial sites were obtained from 25 patients. MCP-1 was associated with CDUS activity (P = 0.009, approximate Spearman's ρ = 0.41), RAMRIS BME score (P = 0.01, approximate Spearman's ρ = 0.42) and RAMRIS erosion score (P = 0.03, approximate Spearman's ρ = 0.31). IL-6 was associated with RAMRIS synovitis score (P = 0.04, approximate Spearman's ρ = 0.50), BME score (P = 0.04, approximate Spearman's ρ = 0.31) and RAMRIS erosion score (P = 0.03, approximate Spearman's ρ = 0.35). MIP-1β was associated with CDUS activity (P = 0.02, approximate Spearman's ρ = 0.38) and RAMRIS synovitis scores (P = 0.02, approximate Spearman's ρ = 0.63). IL-8 associations with imaging outcome measures did not reach statistical significance.. The association between imaging activity and synovial inflammatory mediators underscores the high sensitivity of CDUS and MRI in the evaluation of RA disease activity. The associations found in our present study have different implications for synovial mediator releases and corresponding imaging signs. For example, MCP-1 and IL-6 were associated with both general inflammation and bone destruction, in contrast to MIP-1β, which was involved solely in general synovitis. The lack of association of IL-8 with synovitis was likely underestimated because of a large proportion of samples above assay detection limits among the patients with the highest synovitis scores.

Topics: Adult; Aged; Arthritis, Rheumatoid; Biomarkers; Chemokine CCL2; Chemokine CCL4; Cross-Sectional Studies; Female; Hand Joints; Humans; Immunoassay; Inflammation Mediators; Interleukin-6; Interleukin-8; Magnetic Resonance Imaging; Male; Middle Aged; Radiography; Reproducibility of Results; Sensitivity and Specificity; Synovial Membrane; Synovitis; Tissue Culture Techniques; Ultrasonography, Doppler, Color

Platelet microparticles (pMPs) are small membrane-coated vesicles that are released from the plasma membrane upon platelet activation. In the joint fluid of patients with rheumatoid arthritis, pMP can interact with and activate fibroblast-like synoviocytes (FLS), which are important effector cells that mediate both immune activation and joint destruction. The signaling process by which engagement of glycoprotein VI (GPVI), a surface glycoprotein receptor for collagen which is expressed on platelets, triggers pMP generation is poorly understood, but has been suggested to involve Spleen Tyrosine Kinase (SYK), best known as an upstream activator of Bruton's Tyrosine Kinase (BTK) in B cells. In this study, we showed that activation of human platelets triggered by convulxin or collagen, specific ligands for GPVI receptor, or alternatively by antibody-mediated cross-linking of another platelet receptor, C type lectin-like receptor 2 (CLEC2), resulted in phosphorylation of BTK and downstream effector, phospholipase Cγ2 (PLCγ2). A potent and selective BTK inhibitor, RN486, inhibited GPVI- or CLEC2-mediated PLCγ2 phosphorylation and pMP production in a dose-dependent manner. BTK is also an essential effector of B cell receptor (BCR)-induced B cell signaling. Consistent with the biology, the IC50s of BTK inhibitors with varying potencies in a BCR-dependent B cell activation marker assay correlated with those in the GPVI-mediated PLCγ2 phosphorylation. In a co-culture system consisting of human primary synovial FLS and activated human platelets, convulxin stimulation resulted in elevated production of pro-inflammatory cytokines, IL-6 and IL-8, an effect which was dose-dependently blocked by RN486. The effects are specific as RN486 abrogated platelet aggregation induced by GPVI ligands but not by other platelet surface receptor agonists. Taken together, our data further support the potential therapeutic utility of BTK inhibitors in RA therapy, by inhibiting GPVI-mediated platelet activation and thus subsequent amplification of inflammation driven by pMP-induced FLS cytokines production.

Topics: Agammaglobulinaemia Tyrosine Kinase; Arthritis, Rheumatoid; B-Lymphocytes; Blood Platelets; Catalysis; Cell-Derived Microparticles; Coculture Techniques; Humans; Interleukin-6; Interleukin-8; Lectins, C-Type; Lymphocyte Activation; Phospholipase C gamma; Phosphorylation; Platelet Activation; Platelet Aggregation; Platelet Membrane Glycoproteins; Protein Kinase Inhibitors; Protein-Tyrosine Kinases; Receptors, Antigen, B-Cell; Synovitis

Interleukin (IL)-17A and Th17 cells are critically involved in T cell-mediated synovial inflammation. Besides IL-17A, Th17 cells produce IL-22. Recently, Th22 cells were discovered, which produce IL-22 in the absence of IL-17. However, it remains unclear whether IL-22 and Th22 cells contribute to T cell-mediated synovial inflammation. Therefore, we examined the potential of IL-22 and Th22 cells to induce synovial inflammation and whether IL-22 is required for T cell-mediated experimental arthritis.. Peripheral and synovial Th17 and Th22 cells were identified and sorted from patients with rheumatoid arthritis (RA). Co-culture experiments of these primary T cell populations with RA synovial fibroblasts (RASF) were performed. The in vivo IL-22 contribution to synovial inflammation was investigated by inducing T cell-mediated arthritis in IL-22 deficient mice and wild-type mice.. Peripheral Th17 and Th22 cell populations were increased in patients with RA and present in RA synovial fluid. In T cell-RASF co-cultures, IL-22 in the presence of IL-17A had limited effects on IL-6, IL-8, matrix metalloproteinase-1 (MMP-1) and MMP-3 production. Furthermore, primary peripheral blood and synovial Th17 cells were more potent in the induction of these factors by RASF compared with Th22 cells. In line with this, similar synovial inflammation and disease severity was found between IL-22 deficient and wild-type mice in T cell-mediated experimental arthritis.. These findings show that IL-17A/Th17 cell-mediated synovial inflammation is independent of IL-22 and Th22 cells. This implies that targeting IL-17A/Th17 cells, rather than IL-22/Th22 cells, should be the focus for treatment of T cell-mediated synovial inflammation.

Topics: Adult; Aged; Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Coculture Techniques; Female; Humans; Interleukin-17; Interleukin-22; Interleukin-6; Interleukin-8; Interleukins; Male; Matrix Metalloproteinase 1; Matrix Metalloproteinase 3; Mice; Mice, Inbred C57BL; Mice, Knockout; Middle Aged; Receptors, CCR6; Synovitis; Th17 Cells; Up-Regulation; Young Adult

The underlying mechanisms for the subsets of self-limiting, intermittent or chronic and deforming arthritis in systemic lupus erythematosus (SLE) are not well understood. We performed a cross-sectional analysis of pro-inflammatory cytokines (IL-1β, IL-2, IL-6, IL-8 and TNF-α) and joint status in 47 SLE patients (79% females, age 42 years, disease duration 8.6 years). All cytokines levels were significantly elevated in SLE patients compared with controls, but only IL-2 and IL-8 levels were higher than in patients with rheumatoid arthritis. SLE patients with ongoing synovitis (19%) and joint deformities (11%) had increased erythrocyte sedimentation rate (ESR), IL-6 and anti-dsDNA Ab levels. IL-6 levels correlated with ESR, anti-dsDNA Ab and haemoglobin, but not with C-reactive protein levels. Arthritis constitutes a considerable burden of disease in SLE over time, and joint deformations are associated with longstanding disease and arthritis flare rates. IL-6 is a potential biomarker and therapeutic target in the prevention of joint damage in SLE arthritis.

Topics: Adult; Aged; Arthritis, Rheumatoid; Biomarkers; Blood Sedimentation; Case-Control Studies; Cross-Sectional Studies; Cytokines; Female; Humans; Interleukin-2; Interleukin-6; Interleukin-8; Joint Deformities, Acquired; Lupus Erythematosus, Systemic; Male; Middle Aged; Registries; Synovitis

Bradykinin (BK) and B2 receptors have been implicated in the pathophysiology of osteoarthritis (OA), and synovitis is one of its hallmarks. Here, the selective B2 receptor antagonists MEN16132 and icatibant have been pharmacologically characterized in human synovial cells.. Radioligand and functional studies (inositol phosphate (IP) accumulation, interleukin (IL)-6 and IL-8 release) were performed in cultured synoviocytes.. [3H]-BK saturation studies indicated receptor density (Bmax) and K(d) values of 121,550 sites per cell and 1.14 nM respectively. In synoviocytes, MEN16132 (pK(I) 8.9) was threefold more potent than icatibant (pK(I) 8.4). Both antagonists showed competitive antagonism in the BK-induced IP assay (control EC50 0.45 nM), with pK(B) values of 9.9 (MEN16132) and 8.1 (icatibant). 24h incubation with BK induced IL-6 (EC50 216 nM) and IL-8 (EC50 53 nM) release. Both MEN16132 (IL-6: pIC50 8.1; IL-8: pIC50 8.4) and icatibant (IL-6: pIC50 6.6; IL-8: pIC50 6.7) completely prevented this BK-induced release. Indomethacin did not affect the basal or the IL-6/IL-8 release induced by BK, whereas nordihydroguaiaretic acid decreased the basal release, although BK still increased IL-6 and IL-8 production. BK-induced IL-8 release was attenuated by inhibitors of phospholipase C (U73122), p38 (SB203580), JNK (SP600125), ERK 1/2 (PD98059) MAPKs, phosphoinositide 3-kinase (LY294002), NF-kappaB (BAY-117085) and by the glucocorticoid dexamethasone.. Bradykinin via B2 receptors can participate in inflammatory events in synovitis. MEN16132 is a highly potent B2 receptor antagonist capable of blocking pro-inflammatory responses to BK evoked in human synoviocytes.

Topics: Bradykinin; Bradykinin B2 Receptor Antagonists; Cells, Cultured; Fibroblasts; Humans; Inhibitory Concentration 50; Inositol Phosphates; Interleukin-6; Interleukin-8; Ornithine; Radioligand Assay; Receptor, Bradykinin B2; Sulfonamides; Synovial Membrane; Synovitis

Compressive stress may be involved in temporomandibular joint (TMJ) synovitis, but its mechanism has not been fully elucidated. We hypothesized that mechanical stress to the synovial cells of the TMJ potentially causes degenerative changes in temporomandibular joint disease. We examined the effect of cyclic compressive loading on three-dimensionally engineered constructs using human TMJ synovium-derived cells in vitro. Human TMJ synovium-derived cells were cultured onto collagen scaffolds, resulting in three-dimensional constructs. Cyclic compression loading was applied to the constructs by means of a custom-designed apparatus. DNA amount, apoptotic cells, and mRNA levels for inflammatory cytokines were analyzed. The protein expression and activity of MMPs were examined. DNA amount or apoptotic cell number was unchanged by loading. MMP-2, -3, and IL-8 mRNA expression was up-regulated by the compression, and both MMP-1 and -3 protein expression and MMP-2 activity were detected. Thus, compression of human TMJ synovium-derived cells appears to modulate inflammatory cytokines.

Topics: ADAM Proteins; Adult; Apoptosis; Cell Culture Techniques; Cells, Cultured; Collagen; Compressive Strength; Cytokines; Dental Stress Analysis; Female; Humans; Inflammation Mediators; Interleukin-8; Male; Matrix Metalloproteinase 2; Matrix Metalloproteinase 3; Middle Aged; Synovial Membrane; Synovitis; Temporomandibular Joint Disorders; Tissue Inhibitor of Metalloproteinase-1

Synovitis, which is characterized by infiltration of inflammatory cells, often accompanies progression of clinical symptoms of the temporomandibular joint (TMJ). Synovial fibroblasts of the TMJ are believed to play important roles in progression of synovitis. The purpose of this study was to examine production and gene expression of chemokines by synovial fibroblasts stimulated by tumor necrosis factor-alpha (TNF-alpha).. Protein levels of chemokines were measured by enzyme-linked immunosorbent assay (ELISA). Gene expression of chemokines was analyzed by real-time polymerase chain reaction (PCR).. Production of interleukin (IL)-8, growth-related oncogene (GRO)-alpha, monocyte chemoattractant protein (MCP)-1, and regulated upon activation normal T-cell expressed and secreted (RANTES) protein by synovial fibroblasts was increased by TNF-alpha. In contrast, stromal cell-derived factor (SDF)-1alpha, macrophage inflammatory protein (MIP)-1alpha and -1beta were not detectable in conditioned media of synovial fibroblasts, with or without TNF-alpha treatment. Increases in gene expression of IL-8, GRO-alpha, MCP-1, and RANTES in response to TNF-alpha treatment were detected.. Increased protein production and gene expression of chemokines by synovial fibroblasts in response to TNF-alpha treatment appears to play an important role in recruitment of inflammatory cells into synovium and the progression of synovitis in the TMJ.

Topics: Adolescent; Adult; Chemokine CCL2; Chemokine CCL3; Chemokine CCL4; Chemokine CCL5; Chemokine CXCL1; Chemokine CXCL12; Chemokines; Chemokines, CXC; Disease Progression; Female; Fibroblasts; Gene Expression Regulation; Humans; Intercellular Signaling Peptides and Proteins; Interleukin-8; Joint Dislocations; Macrophage Inflammatory Proteins; Osteoarthritis; Synovial Membrane; Synovitis; Temporomandibular Joint Disc; Temporomandibular Joint Disorders; Tumor Necrosis Factor-alpha

Studies indicate the genetic, biological, and clinical heterogeneity of rheumatoid arthritis (RA). Recently the histological diversity of RA has been postulated. We investigated whether serum concentrations of interleukin 8 (IL-8), RANTES (regulated upon activation normal T cell expressed and secreted), and monocyte chemoattractant protein-1 (MCP-1) are correlated with histological appearance of the rheumatoid synovitis.. Using ELISA we assessed IL-8, RANTES, and MCP-1 concentrations in serum of 47 patients with RA and 30 patients with osteoarthritis (OA).. Morphological analysis of synovial specimens distinguished 2 types of rheumatoid synovitis. Twenty-eight RA samples presented diffuse infiltrates of mononuclear cells with no specific microanatomical organization and were categorized as diffuse synovitis. In the remaining 19 specimens, classified as follicular synovitis, formation of lymphocytic follicles with germinal center-like structures was observed. Serum levels of studied chemokines were increased in patients with RA compared to the OA control group (p < 0.001 for all comparisons). Concentrations of IL-8, RANTES, and MCP-1 were highest in serum of RA patients with follicular synovitis in comparison with patients with diffuse synovitis (p < 0.01, p < 0.01, and p < 0.05, respectively) and could distinguish RA patients with these 2 histological disease patterns. Serum levels of chemokines correlated with markers of disease activity such as erythrocyte sedimentation rate, C-reactive protein concentrations, and Disease Activity Score.. Distinct histological variants of rheumatoid synovitis associated with different serum levels of IL-8, RANTES, and MCP-1 reflect clinical activity of the disease and confirm the concept of RA heterogeneity.

Topics: Adult; Aged; Arthritis, Rheumatoid; Biomarkers; Biopsy, Needle; Case-Control Studies; Chemokine CCL2; Chemokine CCL5; Chemokines; Disease Progression; Female; Humans; Immunohistochemistry; Interleukin-8; Male; Middle Aged; Probability; Prognosis; Reference Values; Risk Assessment; Sensitivity and Specificity; Severity of Illness Index; Synovitis

Monosodium urate monohydrate (MSU) crystals promote gouty inflammation that is critically mediated by neutrophil recruitment and activation. Interleukin-8 (IL-8) and closely related chemokines are major neutrophil chemotaxins in experimental gout. But MSU crystals also activate the classical and alternative pathways of complement, and MSU crystals directly cleave C5 on the crystal surface. Unlike IL-8, the roles in acute gout of individual complement-derived peptides and of the terminal C5b-9 complement components that comprise the membrane attack complex (MAC) are unclear. Hence, we studied rabbits deficient in the MAC component C6 to determine if MAC mediated urate crystal-induced arthritis.. We injected C6-deficient and C6-sufficient rabbit knee joints with 10 mg of pyrogen-free urate crystals and analyzed IL-8 levels, leukocyte influx, and joint inflammation 24 hours later.. There was a significant decrease (>60%) in swelling in MSU crystal-injected knees of C6-deficient animals as compared with C6-sufficient animals (P < 0.05). An attenuated rise in MSU crystal-induced joint effusion levels of IL-8 also was observed, which was concordant with diminished numbers of neutrophils (P < 0.05) but not monocytes in MSU crystal-induced knee synovial fluid from C6-deficient animals. Synovial tissue analysis confirmed mononuclear leukocyte infiltration in response to MSU crystal injection in all animals, but substantial neutrophil infiltration only in C6-sufficient animals.. MAC activation appears to play a major role in intraarticular IL-8 generation and in neutrophil recruitment in experimental acute gouty arthritis of the rabbit knee. C6 and MAC activation may represent novel therapeutic targets for suppression of neutrophil-mediated joint inflammation in gout.

Topics: Animals; Complement C6; Complement Membrane Attack Complex; Crystallization; Gout; Interleukin-8; Knee Joint; Neutrophils; Rabbits; Synovitis; Uric Acid

To investigate concentrations of proinflammatory cytokines in the sera and their mRNA expression in biopsy specimens of evanescent rash and synovitis from patients with active untreated adult onset Still's disease (AOSD).. We measured serum levels of interleukin 6 (IL-6), IL-8, and tumor necrosis factor (TNF-alpha) by immunochemiluminescence method and serum IL-18 levels by ELISA in 50 patients with active untreated AOSD, 20 patients with active rheumatoid arthritis (RA), and 20 healthy controls. Multivariate analysis was used to evaluate the correlation between serum cytokine levels and disease activity and clinical features of AOSD. We also evaluated the expression of cytokine transcripts by real-time quantitative polymerase chain reaction in biopsy specimens of evanescent rash and synovitis from 12 patients with active untreated AOSD.. Significantly higher levels of IL-6, IL-8, IL-18, and TNF-alpha in sera were found in patients with active untreated AOSD compared to healthy controls. Serum levels of IL-6 and IL-18 correlated well with clinical activity score of AOSD patients. Multiple logistic regression analysis showed that serum IL-6 level was a possible predictor for the occurrence of evanescent rash (p = 0.0593), serum IL-8 level was a significant predictor of persistent arthritis, and serum IL-18 level predicted occurrence of liver dysfunction. The levels of mRNA expression of IL-6, IL-18, and IL-8 were significantly higher in the biopsy tissue of Still's rash from AOSD patients compared with those in controls. Levels of mRNA expression of IL-18, IL-8, and TNF-alpha were significantly higher in the synovial membranes of AOSD patients compared with those in osteoarthritis controls. Significantly lower levels of TNF-alpha and IL-8 were found in the sera and in the synovial membranes of AOSD patients compared with those in RA patients. AOSD patients who had a chronic articular course had significantly higher levels of serum IL-8 compared with those who had a monocyclic systemic course.. Significantly higher levels of IL-6, IL-8, IL-18, and TNF-alpha were seen in both sera and pathological tissues of patients with active AOSD. The associations between levels of cytokine profile and distinct clinical manifestations and various patterns of disease course suggest the heterogeneity of pathogenesis in AOSD.

Topics: Adult; Arthritis, Rheumatoid; Cytokines; Exanthema; Female; Humans; Interleukin-18; Interleukin-6; Interleukin-8; Male; Osteoarthritis; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Still's Disease, Adult-Onset; Synovitis; Tumor Necrosis Factor-alpha

We sought to clarify the nature of joint effusion (JE) on T2-weighted magnetic resonance images of the temporomandibular joint (TMJ) by analysis of the synovial fluid in the superior compartment of patients with internal derangement and osteoarthrosis.. One hundred symptomatic TMJs (100 patients) with 65 internal derangements and 35 osteoarthroses were scanned by means of magnetic resonance imaging, and, the synovial fluid was sampled on the same day. The amount of JE was evaluated on a scale of 0 to 3. Grades 0 and 1 indicated absence of JE or a negligible amount of JE, respectively, and grades 2 and 3 indicated the presence of JE. Correlation was evaluated among the amount of JE and the concentrations of the total protein and interleukin-1beta(IL-1beta), IL-6, IL-8, and tumor necrosis factor-alpha in the synovial fluid.. Magnetic resonance imaging revealed the absence of JE in 40 joints (grade 0, 17 joints; grade 1, 23 joints) and the presence of JE in 60 joints (grade 2, 31 joints; grade 3, 29 joints). The joints with JE had, on average, significantly higher concentrations of total protein (1,675 microg vs 714 microg; P = .0001) and IL-6 (42.9 pg vs 10.6 pg; P = .009) than did the joints without JE. Furthermore, there were significant correlations between the JE grade and the concentrations of the total protein (P = .0001), IL-6 (P = .001), and IL-8 (P = .004). The detection ratio of cytokines among the presence-absence groups of JE showed a significant difference in tumor necrosis factor-alpha (68.3% vs 47.5%; P = .037) and IL-6 (86.7% vs 67.5%; P = .012). Conclusions. JE may contain the released products when there is pronounced synovitis. It is probably composed of high concentrations of total protein with inflammatory cytokines. Furthermore, IL-6 and IL-8 seem to have an important role in the pathogenesis of JE in TMJ disorders.

Topics: Adolescent; Adult; Aged; Arthralgia; Chi-Square Distribution; Cytokines; Female; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Joint Dislocations; Magnetic Resonance Imaging; Male; Mandibular Condyle; Middle Aged; Osteoarthritis; Pain Measurement; Proteins; Range of Motion, Articular; Retrospective Studies; Statistics, Nonparametric; Synovial Fluid; Synovitis; Temporomandibular Joint; Temporomandibular Joint Disorders; Tumor Necrosis Factor-alpha

This study investigated the correlations between the concentrations of proinflammatory cytokines in synovial fluid and the degree of synovitis on the one hand, and the degree of degeneration of articular cartilage on the other hand, in patients with internal derangement and osteoarthritis of the temporomandibular joint. We measured the concentrations of interleukin-1beta (IL-1beta), tumour necrosis factor-alpha (TNF-alpha), IL-6 and IL-8 in synovial fluid and the degree of arthroscopic synovitis and degeneration of articular cartilage in 37 joints with internal derangement and osteoarthritis. The correlations between the concentration of each cytokine and the score of each arthroscopic feature were analysed statistically. The detection rates of IL-1beta,TNF-alpha, IL-6 and IL-8 were 57%, 78%, 89% and 70%, respectively. There was a positive correlation between the IL-6 concentration and the synovitis score (P = 0.02). Measurement of IL-6 in synovial fluid might be useful as an indicator of the extent of synovitis.

Topics: Adolescent; Adult; Aged; Arthroscopy; Cartilage, Articular; Female; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Interleukins; Joint Dislocations; Male; Middle Aged; Osteoarthritis; Statistics, Nonparametric; Synovitis; Temporomandibular Joint Disc; Temporomandibular Joint Disorders; Tumor Necrosis Factor-alpha

Rheumatoid arthritis is a prevalent example of an inflammatory angiogenic disease, which is mediated by pro-inflammatory and pro-angiogenic cytokines such as tumor necrosis factor (TNF). To evaluate the effect of the potent anti-angiogenic factor endostatin on TNF-induced inflammatory arthritis, we injected an endostatin-expressing lentiviral vector directly into the joints of human TNF-transgenic mice before the onset of disease. Histological analysis of the injected joints 8 weeks later revealed that endostatin reduced blood vessel density within the synovial tissues and an overall mean arthritis index. In vitro and in vivo examination of the potential mechanism by which endostatin inhibited the arthritis revealed that endostatin blocks TNF-induced activation of JNK and JNK-dependent pro-angiogenic gene expression. These data suggest a novel mechanism by which endostatin inhibits angiogenesis, and demonstrates the potential utility of anti-angiogenic gene therapy for treatment of inflammatory arthritis.

Topics: Angiogenesis Inhibitors; Animals; Arthritis, Rheumatoid; Chemokine CCL2; Collagen; Electrophoretic Mobility Shift Assay; Endostatins; Endothelial Growth Factors; Endothelium, Vascular; Gene Transfer Techniques; Genetic Therapy; Humans; Immunoenzyme Techniques; Interleukin-8; JNK Mitogen-Activated Protein Kinases; Joints; Lymphokines; MAP Kinase Kinase 4; Matrix Metalloproteinases; Mice; Mice, Inbred C57BL; Mice, Transgenic; Mitogen-Activated Protein Kinase Kinases; Neovascularization, Pathologic; Peptide Fragments; RNA; Synovitis; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A; Vascular Endothelial Growth Factors

In chronic inflammatory foci, such as the rheumatoid joint, there is enhanced recruitment of phagocytes from the blood into the tissues. Chemokines are strongly implicated in directing the migration of these cells, although little is known regarding the chemokine receptors that could mediate their chemotaxis into the joint tissue. Therefore the objective of the study was to identify chemokine binding sites on macrophages and neutrophils within the rheumatoid synovium using radiolabeled ligand binding and in situ autoradiography. Specific binding sites for CCL3 (macrophage inflammatory protein-1alpha), CCL5 (RANTES), CCL2 (monocyte chemoattractant protein-1) and CXCL8 (IL-8) were demonstrated on CD68+ macrophages in the subintimal and intimal layers. The number and percentage of intimal cells that bound chemokines were greater in inflamed regions compared to noninflamed regions. The intensity of intimal binding varied between chemokines with the rank order, CCL3 > CCL5 > CCL2 > CXCL8. Neutrophils throughout the synovium bound CXCL8 but did not show any signal for binding CCL2, CCL3 or CCL5. Immunohistochemistry showed that both CXCR1 and CXCR2 are expressed by macrophages and neutrophils in the rheumatoid and nonrheumatoid synovia, suggesting that both of these receptors are responsible for the CXCL8 binding. The chemokine binding sites described on phagocytes may be involved in the migration of these cells into the inflamed joint.

Topics: Antigens, CD; Antigens, Differentiation, Myelomonocytic; Autoradiography; Binding Sites; Chemokine CCL2; Chemokine CCL3; Chemokine CCL4; Chemokine CCL5; Chemokines; Humans; Immunoenzyme Techniques; Interleukin-8; Macrophage Inflammatory Proteins; Macrophages; Neutrophils; Radioligand Assay; Synovitis

To measure the levels of the proinflammatory cytokines, interleukin (IL)-1 beta, IL-6, tumor necrosis factor- (TNF) alpha, IL-8, and interferon- (IFN) gamma in synovial fluid samples taken from patients with temporomandibular disorders (TMD).. We studied 6 asymptomatic volunteers and 51 patients with TMD. The IL-1 beta, IL-6, TNF-alpha, IL-8, and IFN-gamma levels in temporomandibular joint synovial fluid were measured using enzyme-linked immunosorbent assay.. Measurable level of at least one cytokine in the synovial fluid was found in 40 (64.5%) of 62 joints in the patients: IL-1 beta and IFN-gamma were each detected in 18 (29.0%) of 62 joints; IL-6 in 13 (21.0%) of 62 joints; IL-8 in 11 (19.3%) of 57 joints; and TNF-alpha in only 5 (8.1%) of 62 joints. None of these cytokines was detectable in the synovial fluid in the control group. Furthermore, there was a strong correlation between the detection of IL-1 beta and pain in the joint area.. These data clearly demonstrate increased levels of several proinflammatory cytokines in certain patients with TMD and suggest that these cytokines may play a role in the pathogenesis of synovitis and degenerative changes of the cartilaginous tissue and bone of the temporomandibular joint.

Topics: Adolescent; Adult; Aged; Cytokines; Female; Humans; Inflammation Mediators; Interferon-gamma; Interleukin-1; Interleukin-6; Interleukin-8; Joint Dislocations; Male; Middle Aged; Osteoarthritis; Statistics, Nonparametric; Synovial Fluid; Synovitis; Temporomandibular Joint Disorders; Tumor Necrosis Factor-alpha

We describe the cellular infiltrate and cytokine profile in sequential synovial membrane biopsies from a patient with acute followed by chronic synovitis after intravesical bacillus Calmette-Guérin (BCG) therapy for an in situ transitional cell carcinoma of the bladder. Histological and immunohistochemical analysis of 3 synovial biopsies were done sequentially over a 9 month period. The patient was HLA-B27 positive, but HLA-DR4 negative, and did not have the "shared epitope." Unlike other cases, this patient's arthritis did not respond initially to nonsteroidal antiinflammatory drugs and was exacerbated by corticosteroid therapy. The synovitis took a neutrophilic form, with marked synovial membrane content of interleukin 8 (IL-8) and tumor necrosis factor alpha (TNF-alpha). It subsequently developed into chronic lymphoplasmacytoid synovitis, similar to rheumatoid arthritis (RA), with decreased IL-8 but continuing IL-1 and TNF-alpha production in the synovial membrane. The synovitis resolved to a fibrotic synovium with residual thickening of the synovial lining layer and continued production of TNF-alpha. Thus, during the evolution of this arthritis, the synovial layer and continued production of TNF-alpha. Thus, during the evolution of this arthritis, the synovial membrane yielded a cellular infiltrate and cytokine content that had marked similarities with that seen in RA; however, the arthritis eventually remitted spontaneously.

Topics: Aged; Arthritis, Reactive; BCG Vaccine; Carcinoma, Transitional Cell; Cytokines; Female; HLA-B Antigens; HLA-DR Antigens; Humans; Immunohistochemistry; Interleukin-1; Interleukin-8; Synovial Membrane; Synovitis; Time Factors; Tumor Necrosis Factor-alpha; Urinary Bladder Neoplasms

Psoriatic arthritis is an inflammatory arthropathy that ultimately can lead to joint destruction. In this study, we investigated the immunophenotypes of the inflammatory cells and the expression of interleukin-8 (IL-8), which is the hallmark chemoattractant cytokine of psoriasis in synovial membranes from patients exhibiting active psoriatic synovitis (n = 9). The tissue samples were examined by immunohistochemistry, Western blot analysis and in situ hybridisation. The inflammatory infiltrate consisted predominantly of CD3+ T lymphocytes, with a higher proportion of CD4+ than CD8+ T lymphocytes in six cases. CD3+ T lymphocytes were focally distributed near small blood vessels and the enlarged synovial intima. CD1+ interdigitating reticulum cells were not detected. CD22+ B lymphocytes and plasma cells were found in small aggregates without KiM4+ follicular dendritic cells. KiM8+ macrophages were located in the synovial intima and were distributed in a diffuse pattern near the synovial lining cells. CD15+ neutrophil granulocytes were detected in four cases. They were preferentially located in the vicinity of blood vessels and the synovial intima. IL-8 was found at a high level in the synovial lining cells and to a lesser extent in cells located in the perivascular areas. Immunofluorescence double staining showed IL-8 to be expressed in KiM8+ multinucleated giant cells, KiM8+ macrophages and CD3+ T lymphocytes. IL-8 receptor A was demonstrated in the synovial lining and in macrophages and lymphocytes. IL-8 was detected by immunoblot analysis of the synovial tissue at 8.4 kD. Employing in situ hybridisation, IL-8 mRNA was strongly and preferentially expressed in the synovial intima, as well as in macrophages and lymphocytes. The immunophenotype of the psoriatic arthritis inflammatory cells shows great similarity to the inflammatory infiltrate found in the synovial tissue of patients with rheumatoid arthritis. The preferential expression of IL-8 and IL-8 mRNA in the enlarged synovial intima and in lymphocytes and macrophages suggests that IL-8 exerts its action through activated mononuclear cells and T lymphocytes. It seems to play a role in regulating leucocyte traffic into the enlarged synovial intima and may contribute to the aggressive synovitis of patients with psoriatic arthritis.

Topics: Adult; Aged; Antigens, CD; Arthritis, Psoriatic; Female; Humans; Immunohistochemistry; Immunophenotyping; In Situ Hybridization; Interleukin-8; Knee Joint; Macrophages; Male; Middle Aged; Receptors, Interleukin; Receptors, Interleukin-8A; RNA, Messenger; Synovial Membrane; Synovitis; T-Lymphocyte Subsets

Topics: Arthritis, Psoriatic; B-Lymphocytes; Humans; Interleukin-8; Synovial Membrane; Synovitis; T-Lymphocytes

Human neutrophils at inflammatory sites may be an important source of the chemotactic cytokines macrophage inflammatory protein 1 alpha (M1P-1 alpha; a C-C chemokine) and interleukin 8 (IL-8; a C-X-C chemokine). In this study, we show that the inflammatory microcrystals monosodium urate monohydrate (MSU) and calcium pyrophosphate dihydrate (CPPD), the major mediators of gout and pseudogout, differentially regulate the production of these two chemokines by human neutrophils. Both MSU and CPPD increased the secretion of IL-8 by neutrophils in a dose- and time-dependent manner, but had no effect on that of MIP-1 alpha. Since inflammatory cytokines are likely to be present in the synovium during crystal-induced inflammation, we examined the interaction between TNF-alpha and GM-CSF and the crystals. Both TNF-alpha and GM-CSF stimulated IL-8 production; however, only TNF-alpha exerted a significant effect on MIP-1 alpha secretion in neutrophils. IL-8 production induced by TNF-alpha and GM-CSF was synergistically enhanced in the presence of MSU or CPPD, whereas MIP-1 alpha secretion induced by TNF was completely inhibited in the presence of either MSU or CPPD. Interestingly, no interaction between the crystals and the inflammatory cytokines was observed with respect to synthesis of the C-X-C chemokine MGSA in neutrophils. These results suggest that the combination of TNF-alpha and GM-CSF with MSU or CPPD will lead to the production of IL-8 by neutrophils and abolish the release of MIP-1 alpha, an event that will theoretically lead to recruitment of neutrophils but not mononuclear cells. These results are in accordance with the pathological state of gout and pseudogout, where the predominant inflammatory cell is the neutrophil.

Topics: Calcium Pyrophosphate; Cells, Cultured; Chemokine CCL3; Chemokine CCL4; Chemokine CXCL1; Chemokines, CXC; Chemotactic Factors; Crystallization; Cytokines; Gene Expression; Gout; Granulocyte-Macrophage Colony-Stimulating Factor; Growth Substances; Humans; Inflammation; Intercellular Signaling Peptides and Proteins; Interleukin-8; Macrophage Inflammatory Proteins; Monokines; Neutrophils; RNA, Messenger; Synovitis; Tumor Necrosis Factor-alpha; Uric Acid

Topics: Arthritis, Rheumatoid; Chronic Disease; Humans; Interleukin-6; Interleukin-8; Synovitis; T-Lymphocytes

Characteristic cellular changes have previously been reported in the bone marrow of patients with rheumatoid arthritis (RA). We investigated the levels of various cytokines in RA bone marrow.. We studied 25 patients with RA (22 women and 3 men) and 10 trauma patients (7 women and 3 men) as non-RA controls. Twelve kinds of cytokines [interleukin (IL)-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-6, IL-7, IL-8, granulocyte colony stimulating factor, granulocyte/macrophage colony stimulating factor, tumor necrosis factor (TNF)-alpha, and TNF-beta] were assayed by ELISA in iliac bone marrow serum (BMS), tibial BMS, and peripheral blood serum.. Markedly elevated levels of IL-6 and IL-8 were detected in iliac BMS, and much lower levels were found in tibial bone marrow and peripheral blood serum. The levels of IL-6 and IL-8 in iliac BMS showed a close relationship to the extent of synovial proliferation.. Iliac bone marrow may be an important site for the production or accumulation of IL-6 and IL-8 in RA, and these cytokines may influence synovial proliferation in patients with polyarthritis.

Topics: Adult; Aged; Arthritis, Rheumatoid; Bone Marrow; Bone Marrow Cells; Female; Granulocyte Colony-Stimulating Factor; Humans; Ilium; Interleukin-6; Interleukin-8; Male; Middle Aged; Synovitis; Tibia

1. The intradermal administration of endothelial IL-8 (IL-8(1-77) or monocyte derived IL-8 (IL-8(1-72) to rabbits produced a concentration-dependent increase in plasma extravasation and an accumulation of polymorphonuclear leukocytes (PMNs) when measured over a 3 h time period. When plasma extravasation and PMN accumulation were measured over a 30 min time period no significant increases in PMN accumulation or plasma extravasation were observed in response to IL-8 alone. However, under these conditions, the addition of prostaglandin E2 (100 pmol) produced a significant potentiation of IL-8-induced plasma extravasation. There was no significant difference between the biological activities of IL-8(1-77) and IL-8(1-72). 2. Plasma extravasation and PMN accumulation induced by IL-8 were inhibited in rabbits pretreated with the monoclonal antibody designated IB4 (1 mg kg-1, i.v.) directed against the common beta chain (CD18) of the leukocyte integrins. 3. The intra-articular administration to rabbits of IL-8(1-77) (1 nmol) resulted 24 h later in the appearance of a mixed population of leukocytes (PMNs and mononuclear cells) in synovial lavage fluid. Biochemical analyses revealed the presence of an increased level of sulphated proteoglycans (sPG) and of the metalloproteinase stromelysin. Pretreatment of rabbits with IB4 (3 mg kg-1, i.v.) inhibited the accumulation of PMNs but had no effect on the mononuclear infiltrate nor on the levels of sPG or stromelysin. 4. The intradermal or intra-articular injection of E. coli-derived endotoxin induced similar inflammatory changes to those observed with IL-8.The possibility that the biological activities of IL-8 were attributable to minor contamination with endotoxin is unlikely for two reasons. Firstly, biological effects of endotoxin were observed at levels greater than that contained in the IL-8 preparation. Secondly,reduction of the endotoxin content of the IL-8 preparation by a factor of 10 did not produce a concomitant reduction in the observed biological activity of the IL-8.

Topics: Animals; Antibodies, Monoclonal; Antigens, CD; CD18 Antigens; Dermatitis; Endotoxins; Escherichia coli; Female; Interleukin-8; Matrix Metalloproteinase 3; Metalloendopeptidases; Neutrophils; Proteoglycans; Rabbits; Synovial Fluid; Synovitis; Therapeutic Irrigation

Increased amounts of interleukin-8 (IL-8) were detected in synovial fluids of patients with active rheumatoid arthritis (RA) by radioimmunoassay (RIA). The concentration of IL-8 correlated directly with the number of infiltrating neutrophils in synovial fluids. To elucidate the role of IL-8 in neutrophil accumulation at the site of synovitis, the in vivo effects of intraarticular injection of recombinant IL-8 (rIL-8) on leukocytes infiltration into the joint space and synovium were examined. Following a single injection of rIL-8 into the knee joint space of rabbits, redness of the joint and limp became apparent after 4 h and were associated with the rapid infiltration of neutrophilic leukocytes into the joint space and synovial tissues. These effects were time dependent, first becoming evident at 1 h and reaching a plateau in 4 h, and also dose dependent, with a minimal effect being elicited by 100 ng per joint. Although neutrophils were present in the greatest number at 4 h, subsequently mononuclear cells accumulated and became apparent in considerable number after 8 h. Synovial lining cells became ovoid, pleomorphic, and multilayered at 24 h. IL-8 had no effect on the breakdown of proteoglycan of articular cartilage. Based on these findings, IL-8 released from monocytes and synovial cells may be an important contributor to leukocyte accumulation and inflammatory events in the joints of RA.

Topics: Animals; Arthritis, Rheumatoid; Cell Movement; Glycosaminoglycans; Humans; Interleukin-1; Interleukin-6; Interleukin-8; Joints; Leukocytes; Osteoarthritis; Rabbits; Recombinant Proteins; Synovial Fluid; Synovitis