keratan-sulfate and Arthritis--Rheumatoid

Topics: Adult; Aggrecans; Arthritis, Rheumatoid; Body Fluids; Cartilage; Child; Chondroitin Sulfate Proteoglycans; Enzyme-Linked Immunosorbent Assay; Extracellular Matrix Proteins; Humans; Joint Diseases; Keratan Sulfate; Lectins, C-Type; Osteoarthritis; Proteoglycans; Synovial Fluid

Rheumatoid arthritis is a systemic disease. Onset of the disease occurs most often between the ages of 20 and 60 years with a peak at 30 and 50 years. The diagnosis of rheumatoid arthritis (RA) is usually based on the revised criteria for the classification of RA. The presence of 4 or more of 7 criteria defines "rheumatoid arthritis". There is today much interest in development of a biochemical "marker" of RA, to detect the early phase of RA. Some approach employs measurement of serum and joint fluid levels of articular cartilage macromolecules, antibodies, and so on. In the section, we would like to describe some materials available now. It is difficult to point out a single material superior to rheumatoid factor to make a diagnosis of RA at the present time.

Topics: Antibodies; Arthritis, Rheumatoid; Biomarkers; Calcium-Binding Proteins; Collagen; Collagen Type II; Humans; Hyaluronic Acid; Keratan Sulfate

Soft-laser therapy has been used to treat rheumatic diseases for decades. The major effects of laser treatment may be dependent not on thermal mechanisms but rather on cellular, photochemical mechanisms. However, the exact cellular and molecular mechanisms of action have not been elucidated.. The aim of this study was to investigate the ex vivo effects of low-level laser treatment (with physical parameters similar to those applied previously) on protein expression in the synovial membrane in rheumatoid arthritis (RA).. Synovial tissues were laser irradiated, and protein expression was analyzed.. Synovial membrane samples obtained from 5 people who had RA and were undergoing knee surgery were irradiated with a near-infrared diode laser at a dose of 25 J/cm(2) (a dose used in clinical practice). Untreated synovial membrane samples obtained from the same people served as controls. Synovial protein expression was assessed with 2-dimensional polyacrylamide gel electrophoresis followed by mass spectrometry.. The expression of 12 proteins after laser irradiation was different from that in untreated controls. Laser treatment resulted in the decreased expression of α-enolase in 2 samples and of vimentin and precursors of haptoglobin and complement component 3 in 4 samples. The expression of other proteins, including 70-kDa heat shock protein, 96-kDa heat shock protein, lumican, osteoglycin, and ferritin, increased after laser therapy.. The relatively small sample size was a limitation of the study.. Laser irradiation (with physical parameters similar to those used previously) resulted in decreases in both α-enolase and vimentin expression in the synovial membrane in RA. Both proteins have been considered to be important autoantigens that are readily citrullinated and drive autoimmunity in RA. Other proteins that are expressed differently also may be implicated in the pathogenesis of RA. Our results raise the possibility that low-level laser treatment of joints affected with RA may be effective, at least in part, by suppressing the expression of autoantigens. Further studies are needed.

Topics: Adult; Aged; Analysis of Variance; Arthritis, Rheumatoid; Autoantigens; Case-Control Studies; Chondroitin Sulfate Proteoglycans; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Female; Ferritins; Heat-Shock Proteins; Humans; In Vitro Techniques; Intercellular Signaling Peptides and Proteins; Keratan Sulfate; Low-Level Light Therapy; Lumican; Mass Spectrometry; Middle Aged; Phosphopyruvate Hydratase; Synovial Membrane; Vimentin

Proteoglycans bearing keratan sulfate (KS), such as aggrecan, are components of the human cartilage extracellular matrix (ECM). However, the role of KS in influencing cartilage degradation associated with arthritis remains to be completely understood. KS side chains of the length found in human cartilage are not found in murine skeletal tissues. Using a murine model of inflammatory polyarthritis and cartilage explants exposed to interleukin-1α (IL-1α), we examined whether administering KS could influence intraarticular inflammation and cartilage degradation. Acute arthritis was induced by intravenous administration of an anti-type II collagen antibody cocktail, followed by an intraperitoneal injection of lipopolysaccharide. This treatment was followed by an intraperitoneal KS administration in half of the total mice to evaluate the therapeutic potential of KS for ameliorating arthritis. To investigate the therapeutic potential ex vivo, we examined cartilage fragility by measuring IL-1α-induced aggrecan release from cartilage explants treated with or without KS. Intraperitoneal KS administration ameliorated arthritis in DBA/1J mice. The aggrecan release induced by IL-1α was less in cartilage explants containing media with KS than in those without KS. Our data indicate that exogenous KS ameliorated arthritis in vivo and suppressed cartilage degradation ex vivo. KS may have important therapeutic potential in the treatment of inflammatory arthritis. The mechanism responsible for this requires further investigation, but KS may become a novel therapeutic agent for treating inflammatory diseases such as rheumatoid arthritis.

Topics: Animals; Arthritis, Experimental; Arthritis, Rheumatoid; Cartilage; Disease Models, Animal; Interleukin-17; Keratan Sulfate; Lymphotoxin-alpha; Mice; Mice, Inbred DBA; Tumor Necrosis Factor-alpha

The measurement of body fluid levels of biochemical markers in joint tissues has begun to provide clinically useful information. Synovial fluid (SF) plays an important role in articular joint lubrication, nutrition, and metabolism of cartilage and other connective tissues within the joint. The purpose of our study was to identify and characterize osteogenic protein 1 (OP-1) in SF from patients with rheumatoid arthritis (RA) or with osteoarthritis (OA) and to correlate levels of OP-1 with those of hyaluronan (HA) and antigenic keratan sulfate (AgKS). SF was aspirated from the knees of patients with either RA or OA and from the knees of asymptomatic organ donors with no documented history of joint disease. The presence of detectable OP-1 in SF was demonstrated by western blots with specific anti-pro-OP-1 and anti-mature OP-1 antibodies. Measurement of levels of OP-1, HA and AgKS was performed using ELISAs. OP-1 was identified in human SF in two forms, pro-OP-1 and active (mature) OP-1--mature OP-1 being detected only in SF from OA patients and RA patients. Levels of OP-1 and HA were higher in RA patients than in OA patients and asymptomatic donors, while the level of AgKS was highest in SF from asymptomatic donors. Statistically significant differences were found between SF levels of OP-1 in RA and OA patients and between SF levels of AgKS among the three groups tested. The SF content of OP-1 tended to correlate positively with HA levels, but negatively with AgKS concentrations. In conclusion, the results of this study suggest that measurement of OP-1 in joint fluid may have value in the clinical evaluation of joint disease processes.

Topics: Arthritis, Rheumatoid; Autoantigens; Bone Morphogenetic Protein 7; Bone Morphogenetic Proteins; Humans; Hyaluronic Acid; Keratan Sulfate; Knee Joint; Osteoarthritis; Reference Values; Synovial Fluid

Thrombin is a key factor in the stimulation of fibrin deposition, angiogenesis, proinflammatory processes, and proliferation of fibroblast-like cells. Abnormalities in these processes are primary features of rheumatoid arthritis (RA) in synovial tissues. Tissue destruction in joints causes the accumulation of large quantities of free hyaluronic acid (HA) in RA synovial fluid. The present study was conducted to investigate the effects of HA and several other glycosaminoglycans on antithrombin, a plasma inhibitor of thrombin. Various glycosaminoglycans, including HA, chondroitin sulfate, keratan sulfate, heparin, and heparan, were incubated with human antithrombin III in vitro. The residual activity of antithrombin was determined using a thrombin-specific chromogenic assay. HA concentrations ranging from 250 to 1000 mug/ml significantly blocked the ability of antithrombin to inhibit thrombin in the presence of Ca2+ or Fe3+, and chondroitin A, B and C also reduced this ability under the same conditions but to a lesser extent. Our study suggests that the high concentration of free HA in RA synovium may block antithrombin locally, thereby deregulating thrombin activity to drive the pathogenic process of RA under physiological conditions. The study also helps to explain why RA occurs and develops in joint tissue, because the inflamed RA synovium is uniquely rich in free HA along with extracellular matrix degeneration. Our findings are consistent with those of others regarding increased coagulation activity in RA synovium.

Topics: Antithrombin III; Arthritis, Rheumatoid; Autoimmune Diseases; Calcium; Chondroitin Sulfates; Chromogenic Compounds; Dermatan Sulfate; Glycosaminoglycans; Heparin; Heparitin Sulfate; Humans; Hyaluronic Acid; Iron; Keratan Sulfate; Synovial Fluid; Thrombin

We have sought to determine if markers of proteoglycans and collagen type II (CII) degradation can be detected at an early stage following acute knee injury in the synovial fluid (SF) from a group of patients diagnosed with non-infectious knee joint synovitis (KJS). CII, proteoglycans and elastase activity in the SF from patients with KJS were compared to SF from patients with two chronic arthritis conditions: osteoarthritis (OA) and rheumatoid arthritis (RA) as well as normal SF controls.. CII peptides were measured by sandwich ELISA using two monoclonal antibodies: 8:6:D8, a CII-specific antibody, and 14:7:D8 which binds to an amino acid sequence on CII as well as collagens type I, III and V. Epitope 9A4, a neo-epitope resulting from collagenase digestion of CI, CII, and CIII was measured by inhibition ELISA. Proteoglycans measurement included total sulfated glycosaminoglycans (sGAG) by dye-binding assay and 5-D-4 epitope, a keratan sulfate epitope, by inhibition ELISA. Elastase activity was measured colorimetircally using N-succinyl trialanine p-nitroanilide (SANA) substrate.. The quantified CII peptide concentrations by sandwich and inhibition ELISA were significantly higher in SF from patients with KJS (P<0.05) compared to SF from patients with OA, RA and normal aspirates. 5-D-4 and sGAG concentrations were significantly lower (P<0.05) in SF from patients with KJS compared to SF from patients with OA and RA. Elastase activity in SF from patients with KJS and RA were significantly higher (P<0.05) than SF from patients with OA. A significant correlation exists between elastase activity and 9A4 epitope concentration in SF from patients with KJS.. The elevated CII peptides concentrations in KJS SF compared to normal and OA aspirates indicate early signs of cartilage network damage. The low proteoglycans concentrations in SF from patients with KJS may indicate that injury is limited to the superficial zone of cartilage in the patient population studied. The high elastase activity in SF from patients with KJS and RA are linked to the high CII peptides concentration. The elastase activity in the SF from patients with KJS is due to the action of neutrophil elastase (NE) and collagenases, where both contribute to the destruction of the articular cartilage.

Topics: Arthritis, Rheumatoid; Cartilage, Articular; Collagen Type II; Enzyme-Linked Immunosorbent Assay; Epitopes; Humans; Keratan Sulfate; Knee Injuries; Knee Joint; Osteoarthritis, Knee; Pancreatic Elastase; Proteoglycans; Synovial Fluid; Synovitis

To compare synovial fluid (SF) levels of oncostatin M (OSM), tumor necrosis factor alpha (TNFalpha), and interleukin-6 (IL-6) in patients with rheumatoid arthritis (RA) and osteoarthritis (OA) and to determine which correlate best with SF levels of antigenic keratan sulfate (Ag KS), a marker of aggrecan catabolism, and pyridinium crosslinks, markers of the degradation of mature collagen molecules.. SF was drawn from the knee joints of patients with RA (n = 31) or OA (n = 31). Levels of Ag KS, D-pyridinoline (D-Pyr), pyridinoline (Pyr), OSM, TNFalpha, and IL-6 were measured by enzyme-linked immunosorbent assay.. RA patients had higher median SF levels of OSM, TNFalpha, IL-6, and Pyr, but a lower median level of D-Pyr, than OA patients. In both groups, IL-6 levels correlated positively with those of OSM and TNFalpha. However, the correlation between levels of OSM and TNFalpha was only significant in the RA group. Ag KS and Pyr levels correlated positively in RA but not in OA. The correlation between TNFalpha and Ag KS was positive in RA and negative in OA. Further, in RA, OSM and IL-6 levels correlated strongly with Pyr and Ag KS levels but not with D-Pyr levels, while there were no strong correlations in OA for OSM or IL-6 levels with Pyr, Ag Ks, or D-Pyr levels.. This in vivo study suggests that TNFalpha and other proinflammatory cytokines are involved in the up-regulation of the coordinated degradation of cartilage aggrecan and collagen in RA. Further, OSM may act synergistically with other proinflammatory cytokines in up-regulating the production of metalloproteinases by chondrocytes in rheumatoid joints.

Topics: Adult; Aged; Aggrecans; Antigens; Arthritis, Rheumatoid; Biodegradation, Environmental; Biomarkers; Cartilage; Collagen; Cross-Linking Reagents; Cytokines; Extracellular Matrix Proteins; Female; Growth Inhibitors; Humans; Keratan Sulfate; Lectins, C-Type; Male; Middle Aged; Oncostatin M; Osteoarthritis; Peptides; Proteoglycans; Synovial Fluid; Tumor Necrosis Factor-alpha

To investigate the correlation between synovial fluid (SF) concentrations of metalloproteinase (MMP) -2, -3, and -9, tissue inhibitors of metalloproteinases (TIMP)-1 and -2, chondroitin 4-, 6-sulfate (deltadi-C4S, deltadi-C6S), hyaluronic acid (deltadi-HA), antigenic keratan sulfate (KS), and radiologic grade in patients with rheumatoid arthritis (RA) and to assess the clinical value of these factors as disease markers.. Enzyme linked immunoassays and high performance liquid chromatography were used. SF samples were collected from 52 patients with RA. Radiographic (Larsen grade) and clinical evaluations were also done.. There was no significant correlation between the concentrations of MMP, TIMP, and deltadi-C4S, deltadi-C6S, deltadi-HA, and antigenic KS. No significant correlations were found between the radiologic grade and the concentrations of MMP or TIMP in SF. There was a significant increase in the concentration of antigenic KS and deltadi-C6S/deltadi-C4S ratio in SF from patients with Larsen grade 2 compared to grade 5. A significant correlation between deltadi-C6S and antigenic KS concentrations in SF was observed.. Although there were large variances between the samples in this study, proteoglycan metabolism in the cartilage matrix of the patients with RA might have been increased in the early phases of tissue destruction and decreased in advanced stages because of scanty cartilage tissue. The concentrations of MMP-2, -3, -9 and TIMP-1, -2 in SF did not seem to be influenced by the amount of residual cartilage.

Topics: Aged; Arthritis, Rheumatoid; Chondroitin Sulfates; Female; Glycosaminoglycans; Humans; Hyaluronic Acid; Keratan Sulfate; Male; Metalloendopeptidases; Middle Aged; Radiography; Synovial Fluid; Tissue Inhibitor of Metalloproteinases

To evaluate whether and how moderate physical activity following a night of rest influences serum levels of matrix metalloproteinase 3 (MMP-3), tissue inhibitor of metalloproteinases 1 (TIMP-1), antigenic keratan sulfate (Ag KS), and hyaluronan (HA) in 10 normal subjects and 38 patients with rheumatoid arthritis (RA).. Blood was obtained from 20 RA patients before they arose from a night's sleep, and again 1 and 4 hours after they had begun to perform moderate physical activity. Another 18 RA patients remained in bed and blood was sampled at the same time periods. Serum levels of MMP-3, TIMP-1, Ag KS, and HA were measured by enzyme-linked immunosorbent assay. Clinical activity was evaluated by the Lansbury index.. Both in normal subjects and in RA patients who did not remain in bed throughout the period of blood sampling, levels of HA, Ag KS, and MMP-3 increased significantly during the first hour after the subjects arose: the increase in HA and Ag KS correlated with the Lansbury index in the RA group. Three hours later, levels of Ag KS had dropped to baseline values in both groups of subjects. Levels of HA remained significantly and moderately elevated in the RA group but not in the control group, while levels of MMP-3 did not drop significantly in either group. In contrast, levels of HA, Ag KS, and MMP-3 did not change significantly in RA patients who had remained in bed. Unlike the other markers, the levels of TIMP-1 remained unchanged at the different time periods in all 3 groups studied.. Significant changes in serum levels of some metabolic markers occur during the first hour after one arises from a night of sleep, especially in patients with RA. Measurement of the magnitude of these changes at different times in individual patients provides very different information about metabolic changes occurring in joint tissue than does measurement of the level of the markers at a single time point, as is usually currently reported.

Topics: Adult; Antigens; Arthritis, Rheumatoid; Bed Rest; Biomarkers; Exercise; Female; Humans; Hyaluronic Acid; Keratan Sulfate; Kidney Function Tests; Liver Function Tests; Male; Matrix Metalloproteinase 3; Middle Aged; Time Factors; Tissue Inhibitor of Metalloproteinase-1

To determine whether patients with rheumatoid arthritis (RA) express cellular immunity to the purified G1 globular domain of cartilage proteoglycan (PG) aggrecan and whether it is influenced by the removal of keratan sulfate (KS) chains from the molecule.. The G1 globular domain of PG was purified from mature bovine articular cartilage, digested with keratanase, and used in proliferation assays with peripheral blood lymphocytes (PBL) isolated from 43 patients with RA, 11 patients with nonarticular rheumatism (NAR), including soft tissue rheumatism and mechanical back pain, and 13 healthy age- and sex-matched control subjects.. Removal of KS chains from the G1 globular domain resulted in significantly increased prevalence and values of cellular immune responses to G1 in RA patients compared with the control and NAR groups. In the majority of RA patients, KS chains on G1 significantly inhibited its immune recognition by PBL. There was no significant effect of KS removal on the immunity to G1 in patients with NAR and in the healthy control group.. These results reveal that immune reactivity to the G1 globular domain of the cartilage PG aggrecan is enhanced in patients with RA but only when KS chains are removed. Thus, KS chains inhibit immune responses to this domain of aggrecan. Since immunity to the G1 globular domain of aggrecan induces an erosive polyarthritis in BALB/c mice after removal of KS chains, immunity to the G1 globular domain, cleaved by proteases to remove KS chains, may play a role in the pathogenesis of RA.

Topics: Adult; Aged; Aged, 80 and over; Aggrecans; Animals; Arthritis, Rheumatoid; Cattle; Extracellular Matrix Proteins; Female; Humans; Immunity, Cellular; Keratan Sulfate; Lectins, C-Type; Male; Middle Aged; Peptide Fragments; Proteoglycans; Reference Values

To determine concentrations of chondroitin sulphate (CS) and keratan sulphate (KS) epitopes, glycosaminoglycans (GAGs) and hyaluronan (HA) in knee synovial fluid (SF) from normal subjects and patients with osteoarthritis (OA) or rheumatoid arthritis (RA), to test whether these variables may be used as markers of the OA process.. OA was subdivided into large joint OA (LJOA), nodal generalised OA (NGOA), and OA with calcium pyrophosphate crystal deposition (CPA). Clinical assessment of inflammation (0-6) was undertaken on OA and RA knees. Knee SF was examined by enzyme linked immunosorbent assay for: CS epitopes, using monoclonal antibodies 3-B-3 and 7-D-4; KS epitope using monoclonal antibody 5-D-4; and HA, using biotinylated HA binding region of cartilage proteoglycan. Total sulphated GAGs were measured by dye binding with 1:9 dimethylmethylene blue.. Increased SF 3-B-3 concentrations and 3-B-3/GAG ratio were found in OA, compared with RA or normal knees, with higher 3-B-3 and 3-B-3/GAG in LJOA and NGOA than in CPA. SF 7-D-4 and 7-D-4/GAG were reduced in RA, compared with normal and OA; SF 5-D-4 was reduced in OA compared with normal. GAG and HA concentrations were decreased in both OA and RA. No correlations with radiographic scores were observed, but SF 7-D-4 was lower in 'inflamed' compared with 'non-inflamed' RA and OA knees. In patients with bilateral samples there were strong correlations between right and left knees for all SF variables.. Changed concentrations of SF CS and KS can be detected in OA with a profile that differs from that seen in RA. Clinical subgrouping and local joint inflammation may influence these measures, supporting different pathogenesis within OA subgroups and requirement for careful patient characterisation in SF studies.

Topics: Adult; Aged; Arthritis, Rheumatoid; Biomarkers; Chondroitin Sulfates; Enzyme-Linked Immunosorbent Assay; Epitopes; Female; Glycosaminoglycans; Humans; Hyaluronic Acid; Keratan Sulfate; Knee Joint; Male; Middle Aged; Osteoarthritis; Synovial Fluid

To determine if osteocalcin (OC) is locally produced in the joint and to study the relation between markers of bone, cartilage, and synovial tissue turnover.. The concentrations of OC, keratan sulphate epitope (5D4), and hyaluronate (HA) were measured in paired serum and synovial fluid in 10 healthy volunteers and 15 patients with osteoarthritis (OA) and 16 with rheumatoid arthritis (RA). OC was measured with a commercial immunoradiometric assay and concentrations of 5D4 and HA were measured using enzyme linked immunosorbent inhibition assays.. Synovial fluid OC was found to be significantly lower than serum (p < 0.001) in all patients and controls. Synovial fluid OC concentrations were directly correlated with serum concentrations (r = 0.63, p < 0.001) and with age (r = 0.48, p < 0.01). There were also some relations between OC, HA, and 5D4 in patients with OA and RA. The OC concentrations were directly correlated with HA (r = 0.68, p < 0.01) in OA serum and there was a similar correlation in RA synovial fluid (r = 0.69, p < 0.01). A weak negative correlation was found between OC and 5D4 in OA serum (r = -0.55, p = 0.035) while a weak positive correlation was found in RA serum (r = 0.53, p = 0.034).. These results show that more OC is present in the circulation than in knee joint fluids suggesting that synovial fluid OC may be derived from the blood.

Topics: Adult; Aged; Aged, 80 and over; Arthritis, Rheumatoid; Biomarkers; Bone Remodeling; Cartilage, Articular; Female; Humans; Hyaluronic Acid; Immunoradiometric Assay; Keratan Sulfate; Knee Joint; Male; Middle Aged; Osteoarthritis; Osteocalcin; Rheumatic Diseases; Statistics, Nonparametric; Synovial Fluid

This study investigated the synovial fluid concentrations of glycosaminoglycan (GAG), keratan sulphate (KS) epitope 5D4 and chondroitin sulphate (CS) sulphation patterns in healthy volunteers and patients with osteoarthritis (OA) and rheumatoid arthritis (RA). Synovial fluids were collected from knee joints of healthy volunteers (n = 24), and patients with OA (n = 28) and RA (n = 29). Concentrations of GAG and the keratan sulphate epitope 5D4 were measured in 15 of the healthy volunteers, and all of the OA and RA synovial fluids. Total GAG was measured using a dye-binding method and 5D4 by an ELISA. The unsaturated CS disaccharides delta C4 and delta C6 were measured by capillary electrophoresis in all synovial fluids. The concentrations of GAG, 5D4 and delta C6 in the normal synovial fluid were higher but that of delta C4 lower than those of the disease groups. The delta C6:delta C4 ratios correlated with age (r = -0.437, P < 0.001) and the mean value was lower in females than males (2.92 compared with 5.22, P < 0.001). After allowing for age and sex, the delta C6:delta C4 ratio in the control group was significantly elevated (P < 0.001) compared to both OA and RA. The ratio was also related to proteoglycan markers (r = 0.383 for 5D4 and r = 0.357 for GAG). The finding that 5D4 and delta C6:delta C4 ratios are higher in synovial fluid from healthy volunteers compared to OA and RA suggests that they may be markers of the susceptibility of articular cartilage to early damage in arthritis.

Topics: Adult; Age Factors; Aged; Analysis of Variance; Arthritis, Rheumatoid; Biomarkers; Cartilage; Chondroitin; Disaccharides; Epitope Mapping; Female; Glycosaminoglycans; Humans; Keratan Sulfate; Linear Models; Male; Middle Aged; Osteoarthritis; Sex Factors; Sulfur; Synovial Fluid

Recent studies have implicated leukemia inhibitory factor (LIF) in human joint disease. LIF is produced by cultured synovial cells and articular chondrocytes, stimulates cartilage and bone resorption, and has been detected in inflammatory exudates from arthritic joints. The aim of this study was to evaluate the effect of intraarticular injections of human recombinant LIF in the goat. Endotoxin-free, sterile normal saline containing 1 micrograms recombinant human LIF (rhLIF) was injected into the right radiocarpal joints (RCJs) of eight angora goats. The left RCJs were injected with an equivalent volume of vehicle alone (n = 6) or vehicle containing 1 micrograms human albumin (n = 2). Goat joints were examined for clinical features of inflammation, and synovial fluid (SF) was aspirated on days 0, 2, and 6 postinjection. Leukocyte counts and concentrations of keratan sulfate, IL-1 beta, and TNF-alpha were determined in the SF. Proteoglycan synthesis was determined ex vivo in cartilage explants obtained on day 6 postinjection. A statistically significant increase in joint swelling and effusion volume was observed in LIF-injected joints but not in control joints. In the LIF-injected RCJs, the leukocyte count increased from 82 +/- 9 cells/microliters before injection to 10,300 +/- 3357 cells/microliters at day 2 postinjection (p < 0.005) and declined to 678 +/- 113 cells/microliters at day 6 postinjection. Polymorphonuclear leukocytes and monocyte/macrophages predominated in the infiltrate. No appreciable change in leukocyte counts was observed in control joints.(ABSTRACT TRUNCATED AT 250 WORDS)

Topics: Animals; Arthritis, Rheumatoid; Cartilage, Articular; Evaluation Studies as Topic; Extremities; Goats; Growth Inhibitors; Injections, Intra-Articular; Interleukin-6; Keratan Sulfate; Leukemia Inhibitory Factor; Leukocyte Count; Leukocytes; Lymphokines; Male; Proteoglycans; Recombinant Proteins; Synovial Fluid

To measure serum levels of matrix metalloproteinase-1 (MMP-1), matrix metalloproteinase-3 (MMP-3), and tissue inhibitor of MMP-1 (TIMP-1) in patients with rheumatoid arthritis (RA) and in age-matched control subjects, and to determine how these correlate with serum levels of antigenic keratan sulfate (KS) and other biochemical and clinical indicators of disease activity.. Immunoassays were used to measure levels of MMP-1, MMP-3, TIMP-1, and antigenic KS. Radiologic and functional joint scores were based upon Steinbrocker's criteria. Erythrocyte sedimentation rates (ESR) and levels of C-reactive proteins (CRP) were measured.. In RA patients, levels of MMP-3 and TIMP-1 were significantly increased, and strongly correlated with the ESR and CRP levels but not with radiologic or functional joint scores. Levels of antigenic KS were significantly lower in RA patients and correlated negatively with systemic parameters of inflammation and serum levels of TIMP-1.. The increase in serum levels of MMP-3 and TIMP-1 appears to reflect systemic inflammation in RA. The inverse correlation between serum levels of TIMP-1 and antigenic KS suggests that an upregulation of TIMP-1 synthesis might be responsible for the apparent suppression of cartilage aggrecan catabolism in patients with severe inflammatory changes.

Topics: Adult; Aged; Antigens; Arthritis, Rheumatoid; Arthrography; Biomarkers; Blood Sedimentation; C-Reactive Protein; Collagenases; Erythrocyte Count; Female; Glycoproteins; Humans; Inflammation; Keratan Sulfate; Kidney Function Tests; Liver Function Tests; Male; Matrix Metalloproteinase 1; Matrix Metalloproteinase 3; Matrix Metalloproteinase Inhibitors; Metalloendopeptidases; Middle Aged; Neoplasm Proteins; Protease Inhibitors; Tissue Inhibitor of Metalloproteinases

The metabolism of the cartilage proteoglycan aggrecan was studied in patients with osteoarthritis (OA, n = 83), rheumatoid arthritis (RA, n = 127), and in controls (n = 117) using monoclonal antibody-based radioimmunoassays for glycosaminoglycans in the serum and synovial fluid (SF) to detect epitope 846 on chondroitin sulfate (probably only on recently synthesized molecules) and a keratan sulfate (KS) epitope AN9PI, present on intact and degraded molecules. Epitope 846 levels were always elevated in SF over serum (mean 38-fold in OA and 8.6-fold in RA) being highest in OA patients with the longest disease duration and greatest loss of cartilage, and lowest in RA joints with high leucocyte counts. Serum levels were more often elevated in RA (56%) than in OA (19%) and probably reflect increased aggrecan synthesis in diseased joints. KS levels were higher in SF than in serum in 69% of patients (up to 2.3-fold); levels were inversely (OA) and directly (RA) related to SF leucocyte counts. Serum KS was reduced in both diseases and in RA was inversely related to both systemic and joint inflammation markers. SF 846 levels were inversely related to SF KS in both diseases. These epitopes may provide a measure of the balance between cartilage synthesis and degradation in these diseases.

Topics: Adult; Aged; Aged, 80 and over; Aggrecans; Arthritis, Rheumatoid; Cartilage; Chondroitin Sulfates; Epitopes; Extracellular Matrix Proteins; Female; Humans; Keratan Sulfate; Lectins, C-Type; Male; Middle Aged; Osteoarthritis; Proteoglycans; Synovial Fluid

To correlate the serum levels of keratan sulfate (KS) in patients with rheumatoid arthritis (RA) with different clinical and radiographic variables.. Serum KS levels were measured in 85 patients with RA and 41 age matched controls. Patients with RA were classified according to their disease activity, the presence of rheumatoid factor, the medication prescribed, and to the severity of the joint radiographic changes.. Patients with RA had significantly (p < 0.02) higher levels of serum KS compared to the healthy controls. More significantly, there was an inverse correlation of the serum KS levels with the disease activity (p = 0.02) and the severity of the radiographic changes (p = 0.012).. Serum KS levels appear to correlate with the severity of articular cartilage damage in RA.

Topics: Aged; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Rheumatoid; Female; Humans; Keratan Sulfate; Male; Methotrexate; Middle Aged; Prednisone; Radiography

The large aggregating proteoglycan from human cartilage, aggrecan, has recently been shown to possess an immunologically detectable domain with close homology to epidermal growth factor (EGF), that is variably expressed by alternative mRNA splicing. Using a competitive ELISA we detected this domain in sera from both patients with RA and normal controls. The EGF-like domain could only be detected after digestion of sera with chondroitinase ABC, which demonstrates its proteoglycan origin. The concentration of the aggrecan EGF-like domain was considerably elevated in sera from patients with RA as compared to the control sera (P < 0.001), and the concentration of the domain increased with age in both the patient (r = 0.58, P < 0.05) and the control (r = 0.66, P < 0.01) group. These results demonstrate that the EGF-like domain of aggrecan could be of value as a marker of RA.

Topics: Adult; Aged; Aggrecans; Arthritis, Rheumatoid; Biomarkers; Cartilage; Chondroitin Lyases; Chondroitin Sulfate Proteoglycans; Enzyme-Linked Immunosorbent Assay; Epidermal Growth Factor; Extracellular Matrix Proteins; Female; Humans; Keratan Sulfate; Lectins, C-Type; Male; Middle Aged; Proteoglycans

The present study was undertaken to identify the cartilage matrix molecules that are bound with intermolecular disulfide bonds to IgG and serum albumin molecules recovered from the articular cartilage of patients with rheumatoid arthritis (RA) or osteoarthritis (OA). The cartilage specimens were extracted sequentially with three changes of neutral buffer, three changes of 6 M guanidine hydrochloride and then partially degraded with bacterial collagenase. The extracted IgG and albumin, along with matrix molecules bound to these proteins, were isolated with affinity chromatography using antibodies to IgG or human serum albumin conjugated to agarose beads. The isolated materials were characterized with sodium dodecyl sulfate polyacrylamide gel electrophoresis and transfer blotting, using specific antibodies to IgG, albumin, and proteoglycans. In the isolated materials, heteropolymers with IgG or albumin were identified. These polymers contained keratan sulfate and less frequently chondroitin-4-sulfate and chondroitin-6-sulfate. These findings identified the keratan sulfate rich proteoglycans, prevalent at the surface of joint cartilage, as the most common cartilage matrix molecules that are covalently bound to IgG or to serum albumin by disulfide bonds in the articular cartilage of patients with RA or OA.

Topics: Arthritis, Rheumatoid; Cartilage, Articular; Chondroitin Sulfates; Humans; Immunoglobulin G; Keratan Sulfate; Osteoarthritis; Serum Albumin

Three fibrocartilages associated with the proximal interphalangeal joint are described--at the attachment of the central slip to bone, within the slip where it passes over the joint, and the volar plate. Material was obtained at surgery following trauma, Dupuytren's disease and rheumatoid arthritis. The fibrocartilages were structurally distinct and immunolabelled differently with monoclonal antibodies to extracellular matrix components. All fibrocartilages from normal and Dupuytren's fingers contained chondroitin and keratan sulphate. Type II collagen was present in all attachment zones, although there was little in rheumatoid fingers. It was also present in the dorsal hood of some normal fingers, but not in pathological specimens or the volar plate. The results show that the fibrocartilages are dynamic tissues whose composition varies according to function and use, and changes in disease.

Topics: Adult; Aged; Aged, 80 and over; Arthritis, Rheumatoid; Cartilage, Articular; Chondroitin Sulfates; Collagen; Dupuytren Contracture; Female; Finger Joint; Humans; Immunohistochemistry; Keratan Sulfate; Male; Middle Aged

To measure serum levels of tumor necrosis factor alpha (TNF alpha) and interleukin-6 (IL-6) in patients with rheumatoid arthritis (RA) and age-matched control subjects and to study how these correlate with serum levels of hyaluronan (HA) and antigenic keratan sulfate (KS) and other biochemical as well as clinical indicators of disease activity.. Immunoassays were used to measure levels of TNF alpha, IL-6, HA, and antigenic KS in the serum of 35 patients with RA and a group of age- and sex-matched control subjects. Clinical disease activity in the RA group was assessed using the Lansbury index. Drug intake was recorded and the erythrocyte sedimentation rate, and levels of fibrinogen, creatinine, bilirubin, alkaline phosphatase, lactate dehydrogenase, and aminotransferase were measured.. Serum levels of TNF alpha, IL-6, and HA were significantly higher in the RA population than in the control group. In patients with RA, serum levels of HA correlated positively with serum levels of TNF alpha and with clinical joint scores, but only weakly with other laboratory parameters of inflammation. Serum levels of antigenic KS correlated negatively with levels of circulating TNF alpha, but much more weakly with other clinical and biochemical parameters of disease activity.. These in vivo data support in vitro studies which have shown that TNF alpha is a potent stimulator of HA synthesis by synovial lining cells. The results strengthen the contention that serum HA may be a unique marker of synovial involvement and inflammation, rather than of only inflammation, in RA.

Topics: Adult; Aged; Aged, 80 and over; Antigens; Arthritis, Rheumatoid; Blood Sedimentation; C-Reactive Protein; Female; Fibrinogen; Humans; Hyaluronic Acid; Interleukin-6; Keratan Sulfate; Kidney; Liver; Lymphokines; Male; Middle Aged; Tumor Necrosis Factor-alpha

Serum concentrations of antigenic keratan sulphate determined by an enzyme linked immunosorbent assay (ELISA) with a monoclonal antibody were studied in patients with rheumatoid arthritis (RA), osteoarthritis, ankylosing spondylitis, other inflammatory diseases, and a large control group of women without arthritis. Mean keratan sulphate concentrations were low in 117 women with RA compared with 227 female control subjects matched for age drawn from a community survey. There were significant correlations between serum keratan sulphate concentrations in patients with RA and serum C reactive protein and the erythrocyte sedimentation rate. Serum keratan sulphate concentrations were also low in 29 men and women with ankylosing spondylitis and 29 patients with arthritis and high concentrations of C reactive protein. In 98 women undergoing an operation for benign breast disease there were decreases in serum keratan sulphate concentrations after the operation which correlated with doses in serum C reactive protein. No differences were found in keratan sulphate concentrations in 137 women with osteoarthritis compared with controls. Within the group with osteoarthritis there were no differences for the various joint groups and there was no obvious correlation with radiographic severity or progression. These findings suggest serum keratan sulphate is unlikely to be useful as a diagnostic marker in osteoarthritis or RA but indicate a role for inflammation in the regulation of cartilage loss.

Topics: Acute-Phase Reaction; Aged; Arthritis, Rheumatoid; Biomarkers; Blood Sedimentation; C-Reactive Protein; Cartilage, Articular; Enzyme-Linked Immunosorbent Assay; Female; Humans; Keratan Sulfate; Male; Middle Aged; Osteoarthritis; Rheumatic Diseases; Spondylitis, Ankylosing

The concentration of keratan sulfate (KS) epitope was measured in the serum of patients with osteoarthritis (OA) or rheumatoid arthritis (RA) by enzyme-linked immunosorbent assay and compared with that in the serum of patients with primary fibromyalgia syndrome (PFS) and of controls who had no joint disease. By Student's tau-test, the mean serum KS concentrations in OA and RA patients measured with monoclonal antibodies (MAb) 5-D-4 and 2-D-3 were significantly increased over those in the PFS and normal groups; similar findings were observed using a nonparametric test, except that levels in RA patients showed no difference from those in PFS patients and normal subjects. There was no significant correlation between joint scores or disease duration and KS levels in OA or RA patients. Gel filtration of sera revealed mainly large, polydisperse KS-bearing fragments which eluted in a broad profile. KS purified from sera by immunoaffinity chromatography consisted mainly of high-density proteoglycans. Electrophoresis of pooled high-density KS fractions in polyacrylamide-agarose gels followed by Western blotting with MAb 5-D-4 showed diffuse bands with relative mobilities corresponding to large proteoglycans. These findings are consistent with attachment of KS to protein core fragments of various sizes; KS in patient sera is comparable in size with that in normal sera. Elevations of serum KS levels occur in the presence of cartilage degradation, but do not quantitatively define the extent or duration of articular involvement.

Topics: Arthritis, Rheumatoid; Blotting, Western; Chromatography, Affinity; Chromatography, Gel; Enzyme-Linked Immunosorbent Assay; Fibromyalgia; Humans; Joints; Keratan Sulfate; Molecular Structure; Osteoarthritis

Serum levels of keratan sulphate (KS) were found to be significantly elevated in patients with destructive and predominantly seronegative rheumatoid arthritis (RA) compared with a control population. Levels in RA did not correlate with clinical or laboratory indices of joint activity or damage. Conversely levels were depressed in ankylosing spondylitis (AS) compared with controls.

Topics: Arthritis, Rheumatoid; Biomarkers; C-Reactive Protein; Humans; Keratan Sulfate; Reference Values; Spondylitis, Ankylosing

A new sandwich-ELISA for the determination of keratan sulphate peptides in biological fluids is described. The technique involves binding a commercially available monoclonal antibody against keratan sulphate to microtitre plates, adding the unknown keratan sulphate antigen then interacting the keratan sulphate-antibody complex with a biotin-monoclonal antibody conjugate which was also specific for keratan sulphate peptides. Alkaline phosphatase conjugated streptravidin was then added and the amount which bound to the biotin was determined by measuring the release of chromogen from an added chromogenic substrate. Using this assay, keratan sulphate peptides in biological fluids within the range 10-1000 ng/ml could be quantitated. This method was found to be more sensitive than presently used techniques. The intra- and inter-assay coefficients of variation were 11% and 13%, respectively.

Topics: Adult; Arthritis, Rheumatoid; Enzyme-Linked Immunosorbent Assay; Humans; Keratan Sulfate; Middle Aged; Osteoarthritis; Peptides; Proteoglycans; Synovial Fluid

Single analyses of peripheral blood of rheumatoid arthritis (RA) patients showed a significant reduction in the mean value for keratan sulfate (KS) compared with that in control subjects, but the mean value for orosomucoid (OM) was elevated compared with that in control subjects. Some RA patients displayed highly elevated levels of hyaluronic acid (HA), while others exhibited normal levels. There was a significant inverse correlation between OM and KS content in RA patients, as well as a direct correlation between HA and OM. In longitudinal studies of RA patients, parallel changes in OM and HA and inverse changes between KS and OM or HA were commonly observed. Clinical analyses revealed that there was an inverse correlation between KS and morning stiffness, and direct correlations between the number of tender joints and HA, and between HA or the erythrocyte sedimentation rate and the number of joints with effusions. The reason(s) for the inverse correlation between KS and OM as an index of systemic inflammation remains to be established. Circulating HA represents an index of joint inflammation, for which a marker has not been previously available.

Topics: Adult; Aged; Aging; Arthritis, Rheumatoid; Biomarkers; Blood Sedimentation; Cartilage; Female; Glycosaminoglycans; Humans; Hyaluronic Acid; Keratan Sulfate; Longitudinal Studies; Male; Middle Aged; Orosomucoid; Reference Values; Sex Characteristics

The effects of the chondroprotective agents (Arteparon, SP-54 and DH40J) on the release of proteoglycan degradation products (as keratan sulphate peptide fragments) from articular cartilage implanted into rat subcutaneous air pouches have been investigated by using an enzyme-linked immunosorbent-inhibition assay (ELISIA). The ELISIA technique was capable of quantitating the keratan sulphate peptides (KS peptides) in fluids within the range of 100-2,000 ng/ml by using the monoclonal antibody line 1/20/5-D-4 and human articular cartilage KS peptides as standard reagents. It was found that the levels of KS peptides present in the air-pouch fluid of rats treated with the chondroprotective drugs was significantly less than in fluid aspirated from the pouches of non-drug-treated control animals. On the basis of these findings we suggest that the assessment of KS peptide by ELISIAs may provide a useful means of monitoring proteoglycan breakdown products in biological fluids (e.g. synovial fluids or blood) and for evaluating the effects that antiarthritic drugs may have on this process.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Arthritis, Rheumatoid; Cartilage, Articular; Chondroitin Sulfate Proteoglycans; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Glycosaminoglycans; Keratan Sulfate; Lumican; Male; Osteoarthritis; Pentosan Sulfuric Polyester; Peptide Fragments; Rats; Rats, Inbred Strains

Immuno-electron microscopy has been utilised to examine the cartilage-pannus junction in seven patients with rheumatoid arthritis. Using a monoclonal antibody, keratan sulphate was localised to cells with the ultra-structural appearance of fibroblasts within a transitional fibroblastic zone in the pannus in two cases. This confirms previous light microscopy evidence that this area (which is clearly separate from articular cartilage) contains cells which produce keratan sulphate, and strengthens the hypothesis that these cells are derived from cartilage rather than from the adjacent synovial membrane.

Topics: Antibodies, Monoclonal; Arthritis, Rheumatoid; Cartilage; Cartilage, Articular; Humans; Immunologic Techniques; Keratan Sulfate; Microscopy, Electron

The intensity of safranin 'O' staining is directly proportional to the proteoglycan content in normal cartilage. Safranin 'O' has thus been used to demonstrate any changes that occur in articular disease. In this study, staining patterns obtained using monoclonal antibodies against the major components of cartilage proteoglycan chondroitin sulphate (anti CS) and keratan sulphate (anti KS), have been compared with those obtained with safranin 'O' staining, in both normal and arthritic tissues. In cartilage where safranin 'O' staining was not detectable, the monoclonal antibodies revealed the presence of both keratan and chondroitin sulphate. Thus, safranin 'O' is not a sensitive indicator of proteoglycan content in diseases where glycosaminoglaycan loss from cartilage has been severe.

Topics: Adult; Aged; Aged, 80 and over; Antibodies, Monoclonal; Arthritis; Arthritis, Rheumatoid; Cartilage, Articular; Chondroitin Sulfates; Extracellular Matrix; Humans; Immunohistochemistry; Keratan Sulfate; Middle Aged; Osteoarthritis; Phenazines; Proteoglycans

Antibodies were used in radioimmunoassays with gel chromatography to detect the hyaluronic acid-binding region, core protein, and keratan sulfate of human cartilage proteoglycan in the synovial fluids of patients with rheumatoid arthritis, juvenile rheumatoid arthritis, and osteoarthritis. All fluids contained proteoglycan that was mainly included on Sepharose CL-4B; this result indicates cleavage of proteoglycan (which is normally excluded). The hyaluronic acid-binding region was the smallest and most commonly detected fragment. It was relatively free of keratan sulfate and core protein, and it could sometimes bind to hyaluronic acid. Other larger fragments containing core protein and/or keratan sulfate were detected in every fluid.

Topics: Aggrecans; Arthritis; Arthritis, Juvenile; Arthritis, Rheumatoid; Carrier Proteins; Cartilage, Articular; Extracellular Matrix Proteins; Glycoproteins; Humans; Hyaluronan Receptors; Keratan Sulfate; Lectins, C-Type; Osteoarthritis; Proteoglycans; Radioimmunoassay; Synovial Fluid

In the cartilage-pannus junction of 14 patients with rheumatoid arthritis (RA) and seven patients with osteoarthritis (OA), monoclonal antibodies to keratan sulphate (KS) and chondroitin sulphate (CS) stained a transitional fibroblastic zone (TFZ) within the pannus in nine RA patients and one OA patient. In three patients this was clearly localised to the cytoplasm of cells in this zone, but in all remaining cases KS and CS could be demonstrated in the surrounding matrix. This area was distinguished from adjacent pannus which contained many blood vessels and cells positive for MHC Class II antigen. Specific markers for glycosaminoglycans have been employed to demonstrate that chondrocyte-derived cells and matrix contribute to the changes seen at the cartilage-pannus junction in RA-affected joints.

Topics: Adult; Aged; alpha 1-Antichymotrypsin; Antibodies, Monoclonal; Arthritis, Rheumatoid; Cartilage, Articular; Chondroitin Sulfates; Female; Histocompatibility Antigens Class II; Humans; Keratan Sulfate; Male; Middle Aged; Osteoarthritis