alpha-chymotrypsin and Arthritis--Rheumatoid

Topics: Amyloidosis; Animals; Arteriosclerosis; Arthritis, Rheumatoid; Blood Coagulation Disorders; Blood Coagulation Factors; Chymotrypsin; Chymotrypsinogen; Diabetes Mellitus; Emphysema; Fibrinolysis; Gastrointestinal Diseases; Humans; Kallikreins; Kidney Diseases; Muscular Dystrophies; Oligopeptides; Peptide Hydrolases; Substrate Specificity; Swine

Topics: Adrenal Cortex Hormones; alpha 1-Antitrypsin; alpha-Macroglobulins; Animals; Anti-Inflammatory Agents; Aprotinin; Arthritis, Experimental; Arthritis, Rheumatoid; Beta-Globulins; Carrageenan; Chymotrypsin; Dimethylnitrosamine; Humans; Inflammation; Kallikreins; Leukocytes; Liver; Lysosomes; Macrophages; Protease Inhibitors; Protein Binding; Proteins; Synovial Fluid

The aim of this study was to examine the inhibitory effect of high molecular weight hyaluronan (HA) on nuclear factor (NF)-kappaB activation by COOH-terminal heparin-binding fibronectin fragment (HBFN-f) in rheumatoid arthritis (RA) chondrocytes. When RA chondrocytes in monolayer or cartilage explants were cultured with HBFN-f, the fragment stimulated the phosphorylation and nuclear translocation of NF-kappaB, leading to nitric oxide (NO) production in association with inducible form of NO synthase (iNOS) up-regulation. Inhibition studies with NF-kappaB inhibitors indicated the requirement of NF-kappaB for HBFN-f-induced NO production. Pretreatment with 2700 kDa HA resulted in significant suppression of NF-kappaB activation by HBFN-f. HA also inhibited HBFN-f-stimulated NO production with down-regulation of iNOS. The present study clearly demonstrated that high molecular weight HA suppressed HBFN-f-activated NF-kappaB in RA chondrocytes. HA could down-regulate the catabolic action of fibronectin fragments like HBFN-f in RA joints as a potent NF-kappaB inhibitor.

Topics: Arthritis, Rheumatoid; Cartilage; Cartilage, Articular; Cell Nucleus; Chondrocytes; Chymotrypsin; Fibronectins; Gene Expression Regulation; Heparin; Humans; Hyaluronic Acid; Models, Biological; NF-kappa B; Nitric Oxide; Protein Binding; Protein Structure, Tertiary

Therapy with oral proteolytic enzymes (OET) with combination drug products containing papain, bromelain, trypsin, and chymotrypsin has been shown to be beneficial in clinical settings such as radiotherapy-induced fibrosis, bleomycin pneumotoxicity and immunosuppression in cancer, all of which are nowadays known to be accompanied by excessive transforming growth factor-beta (TGF-beta) production. It has been demonstrated that proteolytic enzymes reduce TGF-beta levels in serum by converting the protease inhibitor alpha2 macroglobulin (alpha2M) from the "slow" form into the "fast" form, whereby the "fast" form binds and inactivates TGF-beta irreversibly. In this study we have investigated the effect of OET on the concentration of TGF-beta1 in serum of patients with rheumatoid arthritis (RA) (n = 38), osteomyelofibrosis (OMF) (n = 7) and herpes zoster (HZ) (n = 7). Seventy-eight healthy volunteers served as controls. TGF-beta1 levels in serum were assessed by enzyme-linked immunosorbent assay (ELISA). We have demonstrated that in healthy volunteers and in patients there exists a correlation between active and latent TGF-beta1 in serum (r=0.8021; P<0.0001). Treatment with OET had no significant effect on TGF-beta1 concentration in healthy volunteers or patients with a normal level of TGF-beta1. In patients with elevated TGF-beta1 concentration (> 50 ng/ml serum), OET reduced TGF-beta1 in RA (P < 0.005), in OMF (P < 0.05) and in HZ (P < 0.05).. These results support the concept that OET is beneficial in diseases characterized in part by TGF-beta1 overproduction.

Topics: Administration, Oral; Adult; alpha-Macroglobulins; Arthritis, Rheumatoid; Bromelains; Chymotrypsin; Drug Combinations; Endopeptidases; Herpes Zoster; Humans; Papain; Primary Myelofibrosis; Rutin; Transforming Growth Factor beta; Transforming Growth Factor beta1; Trypsin

To characterize the effect of added fibronectin (Fn) on human rheumatoid synoviocytes (RAS).. Early passage cultured RAS were studied by fluorescent imaging microscopy with multiple parameter overlaying and confocal laser scanning microscopy.. Added Fn induced a localized decrease in matrix Fn at sites of cell overlap. Similar matrix depletion could be induced by collagen VI, cell binding (120 k) and heparin binding (60 k) fragments of Fn, but not by gelatin binding fragment (45 k).. The decrease in matrix Fn was associated with the induction of a transformation-like phenotype of overlapping cell growth. Both phenomena were inhibited by serine protease inhibitors.

Topics: Arthritis, Rheumatoid; Cell Division; Cells, Cultured; Chymotrypsin; Collagen; Culture Media; Extracellular Matrix; Fibroblasts; Fibronectins; Humans; Image Processing, Computer-Assisted; Microscopy, Confocal; Middle Aged; Osteoarthritis; Peptide Fragments; Serine Proteinase Inhibitors; Synovial Membrane; Time Factors; Trypsin Inhibitors

The link protein components of proteoglycan aggregates in adult human articular cartilage show heterogeneity due to proteolysis. Cleavages near the N-terminus of the intact link proteins, before residues 17, 19 and 24, generate three proteins of slightly diminished size (LP3). Cleavages within the N-terminal disulphide-bonded loop, before residues 66 and 73 of the intact link proteins, generate proteins that yield smaller degradation products upon reduction (LP fragments). In vitro, modified link protein components of a similar size to LP3 can be generated by a variety of proteinases, but of the physiologically relevant enzymes only stromelysin, cathepsin B and cathepsin G have the ability to yield modified link proteins with N-termini identical with those observed in situ. None of the proteolytic agents tested was able to produce LP fragments with N-termini identical with those observed in situ, and the majority of proteinases were not able to cleave within the disulphide-bonded loops. Cathepsin L and hydroxyl radicals can cleave within the N-terminal disulphide-bonded loop, and have the potential of initially opening the loop to allow further proteolytic processing by other agents to generate the native cleavage sites.

Topics: Adult; Amino Acid Sequence; Arthritis, Rheumatoid; Cartilage, Articular; Cathepsin B; Cathepsin G; Cathepsin L; Cathepsins; Chymotrypsin; Cysteine Endopeptidases; Endopeptidases; Extracellular Matrix Proteins; Humans; Infant, Newborn; Matrix Metalloproteinase 3; Metalloendopeptidases; Middle Aged; Molecular Sequence Data; Osteoarthritis; Peptide Fragments; Proteins; Proteoglycans; Serine Endopeptidases

15 different monoclonal antibodies (mcAbs) have been raised against the cleaved (inactive) form of the serpin alpha 1-antitrypsin (AT). In initial experiments these mcAbs were analysed for their ability to bind the native and the cleaved form of this inhibitor: eight of the 15 mcAbs appeared to react predominantly with cleaved AT. Additional experiments with mixtures of purified native AT, AT complexed to neutrophilic elastase and inactivated AT revealed that all mAbs that preferentially reacted with inactivated AT also bound to complexed AT. Using two of the mcAbs against inactivated AT a quantitative and sensitive sandwich-type radioimmunoassay was developed to determine levels of proteolytically inactivated AT in biological fluids. With this assay increased levels of inactivated AT were found in synovial fluid from patients with rheumatoid arthritis corresponding to about 2.4% (range 0.3-11%) of total AT. Approximately 10% of this inactivated AT appeared to consist of AT complexed to neutrophil elastase. The mcAbs described here further illustrate the structural resemblance between the complexed and cleaved forms of AT. In addition, these mcAbs appear to be useful tools for the study of AT in human disease.

Topics: alpha 1-Antitrypsin; Animals; Antibodies, Monoclonal; Arthritis, Rheumatoid; Chymotrypsin; Cross Reactions; Electrophoresis, Polyacrylamide Gel; Humans; Mice; Mice, Inbred BALB C; Neutrophils; Pancreatic Elastase; Precipitin Tests; Protein Denaturation; Rabbits; Radioimmunoassay; Synovial Fluid

The interaction between fibronectin and C1q was studied in the presence of normal human plasma and rheumatoid synovial fluid by solid phase binding assay. Fibronectin-C1q binding occurred in the presence of rheumatoid synovial fluid but not in the presence of normal plasma. Binding was strongest at 4 degrees C and in the presence of EDTA. Fibronectin-C1q binding could be induced in the presence of normal plasma by hypotonicity, augmentation of the concentration of solution-phase fibronectin or by the addition of heat-aggregated IgG. The C1q present in rheumatoid synovial fluid bound to both aminoterminal collagen-binding and carboxyterminal noncollagen binding fibronectin fragments although binding to the aminoterminal fragment was stronger. The interaction between fibronectin and C1q in rheumatoid synovial fluid may modulate immune-complex deposition and complement activation in the inflamed joint.

Topics: Arthritis, Rheumatoid; Chymotrypsin; Complement Activating Enzymes; Complement C1; Complement C1q; Edetic Acid; Electrophoresis, Polyacrylamide Gel; Fibronectins; Humans; Plasma; Synovial Fluid; Temperature

Tissue inhibitor of metalloproteinases (TIMP) from cultured bovine dental pulp inhibits human rheumatoid synovial matrix metalloproteinase 3 (MMP-3) with a stoichiometry of 1:1 on a molar basis. Among the serine proteinases examined, human neutrophil elastase, trypsin and alpha-chymotrypsin destroyed the inhibitory activity of TIMP against MMP-3 by degrading the inhibitor molecule into small fragments. In contrast, the inhibitory activity of TIMP was not significantly reduced by the actions of cathepsin G, pancreatic elastase and plasmin. These data indicate that neutrophils which infiltrate tissues in various inflammatory conditions may play an important role in regulating TIMP activity in vivo through the action of neutrophil elastase.

Topics: Animals; Arthritis, Rheumatoid; Cattle; Chymotrypsin; Enzyme Inhibitors; Humans; Metalloendopeptidases; Neutrophils; Pancreatic Elastase; Serine Endopeptidases; Synovial Membrane; Tissue Inhibitor of Metalloproteinases; Trypsin

Microheterogeneity of two acute-phase proteins: orosomucoid (alpha1-acid glycoprotein, AGP) and alpha 1-antichymotrypsin (ACHT) were studied in the sera of 48 patients with rheumatoid arthritis and 12 healthy individuals. Each rheumatoid patient was assigned to one of four activity grades. Cross affinoimmunoelectrophoresis (aff-EP) with free Concanavalin A (Con A) revealed three microheterogeneity variants of AGP and four microheterogeneity forms of ACHT. The relative amounts of AGP-variants and ACHT-variants observed in the healthy donors were similar to those observed in the patients with activity grade I, but they changed with the increase in the grade of rheumatoid activity. The differences were most significant for AGP. The comparison of serum C-reactive protein (CRP) levels with AGP-variant-O-non reactive with Con A showed significant correlation in respective rheumatoid activity grades.

Topics: Acute-Phase Proteins; alpha 1-Antichymotrypsin; Arthritis, Rheumatoid; Blood Proteins; Chromatography, Affinity; Chymotrypsin; Concanavalin A; Humans; Immunoelectrophoresis; Orosomucoid; Protease Inhibitors

Thyroxine-binding prealbumin (TBPA) concentrations were measured in 54 patients with rheumatoid arthritis (RA) and 50 control subjects. TBPA levels were significantly depressed in the RA patients, of whom 15 had values below the laboratory reference range. Although significant negative correlations were seen between TBPA and the erythrocyte sedimentation rate, C-reactive protein and alpha 1-antichymotrypsin measurements, TBPA levels showed little relationship to disease activity as assessed clinically. On the other hand, RA patients with reduced TBPA had an increased frequency of associated anthropometric and serum visceral protein abnormalities indicating nutritional impairment. TBPA is probably subjected to diverse stimuli in patients with RA and should not be considered to act as a 'pure' negative acute phase reactant.

Topics: Adult; Aged; alpha 1-Antichymotrypsin; Arthritis, Rheumatoid; Blood Sedimentation; C-Reactive Protein; Chymotrypsin; Female; Humans; Male; Middle Aged; Nutrition Disorders; Prealbumin; Thyroxine-Binding Proteins

Human alpha 1-antichymotrypsin, isolated at pH 8.0 from both normal and acute phase plasma, has been found to have two different amino terminal sequences despite the fact that inhibitory activities are unchanged. In normal plasma over 90% of the protein has an amino terminal sequence beginning with aspartic acid and less than 10% with arginine. However, in acute rheumatoid arthritis plasma 55% of the inhibitor begins with arginine and the remainder with aspartic acid. Sequence studies indicate that a fifteen amino acid peptide fragment has been cleaved to yield the arginine protein. Human alpha 1-proteinase inhibitor also shows this heterogeneity, but the ratios do not change between normal and acute phase plasma. It may well be that the missing peptide has some biological activity manifested only in the acute phase state.

Topics: Acute Disease; alpha 1-Antichymotrypsin; Amino Acid Sequence; Arthritis, Rheumatoid; Chymotrypsin; Humans

Paired plasma and synovial fluids from 17 patients with seropositive rheumatoid arthritis were examined for electrophoretic homogeneity/heterogeneity and enzymic inhibitory capacity of the protease inhibitors. The high degree of saturation (approximately 90%) of the polyvalent protease inhibitor alpha 2-macroglobulin in the rheumatoid synovial fluid contrasts sharply with the low saturation of alpha 1-anti-trypsin. The inhibitory reactivity of the non-complexed fraction of both of these dominating antiproteases was retained (approximately 85-90%). Thus, a selective inactivation of synovial alpha 1-antitrypsin could not be demonstrated. alpha 1-Anti-chymotrypsin revealed electrophoretic homogeneity in all synovial fluids. Electrophoretic heterogeneity of the plasmin inhibitor antiplasmin was detected in the majority of synovial fluids indicating plasmin activation. The existence of a protease-antiprotease imbalance in the rheumatoid joint was indicated by the high degree of saturation of alpha 2-macroglobulin and a cleavage of C3 in rheumatoid synovial fluids.

Topics: alpha 1-Antichymotrypsin; alpha 1-Antitrypsin; alpha-2-Antiplasmin; alpha-Macroglobulins; Arthritis, Rheumatoid; Chymotrypsin; Complement C3; Electrophoresis; Female; Humans; Male; Middle Aged; Protease Inhibitors; Synovial Fluid

The amino acid sequence of the 112 residues from the amino terminus of alpha 2-CB5 from chick skin collagen was determined by automated sequential degradation of intact alpha 2-CB5 and several chymotryptic and tryptic peptides. This segment of the peptide includes the site of the action of animal collagenases. As compared to the sequence around the alpha 1 cleavage site, the alpha 2 sequence is notable for the remarkable constancy of the residues to the amino side and the relative abundance of hydrophobic residues to the carboxyl side of the cleavage site, suggesting that these features are important in the recognition by the enzyme. The sequence of this region of the alpha 2 chain is consistent with the Gly-X-Y triplet structure and the preference of certain residues for either the X or Y position in distribution. However, three of the six residues of leucine were found in the Y position rather than the X position. Leucine residues were found only once in the Y position in the alpha 1 (I) chain. This preference does not appear to hold in the alpha 2 chain.

Topics: Amino Acid Sequence; Animals; Arthritis, Rheumatoid; Chickens; Chymotrypsin; Collagen; Humans; Microbial Collagenase; Peptide Fragments; Skin; Synovial Fluid; Trypsin

Serum alpha1-antichymotrypsin (alpha1-ACT) of the patient with rheumatoid arthritis was studied by means of single radial immunodiffusion method. There was a significant elevation of the alpha1-ACT concentration in the patients with rheumatoid arthritis, and positive relationships were observed between the concentrations of alpha1-ACT and of other glycoproteins such as alpha1-acid glycoprotein and alpha1-antitrypsin in individual patients, and between C-reactive protein (CRP) rates and the (CRP) rates and the alpha1-ACT concentrations in individual specimens. These facts suggest that alpha1-ACT belongs to a group of acute phase proteins like alpha1-antitrypsin or CRP. An inverse proportional correlation was revealed between alpha1-ACT and fibrinolytic activity. No influences were observed on the alpha1-ACT concentration, activity index or articular index by the oral administration of alpha-chymotrypsin in patients with rheumatoid arthritis.

Topics: Adult; alpha 1-Antitrypsin; Arthritis, Rheumatoid; C-Reactive Protein; Chymotrypsin; Female; Fibrinolysis; Glycoproteins; Humans; Male

Topics: alpha-Macroglobulins; Antithrombins; Arthritis, Rheumatoid; Chymotrypsin; Humans; Protease Inhibitors

The concentrations of five normally occurring protease inhibitors in serum and synovial fluid were compared in patients with rheumatoid arthritis, osteoarthrosis, and normal controls. The patients with rheumatoid arthritis showed a significant rise in alpha1-antitrypsin, alpha1-antichymotrypsin, and inter-alpha-trypsin inhibitor (in decreasing order) in serum as well as in synovial fluid. In synovial fluid the inhibitors were present in their native form and bound to hyaluronate. A large molecular protein with immunological specificity of alpha1-antitrypsin, presumably a complex of alpha1-antitrypsin and a protease, could be shown in synovial fluid of all patients with classical and probable rheumatoid arthritis and not in that of the other subjects studied.23Author

Topics: alpha 1-Antitrypsin; Arthritis, Rheumatoid; Chromatography, Gel; Chymotrypsin; Female; Humans; Hyaluronoglucosaminidase; Immunoelectrophoresis; Male; Molecular Weight; Osteoarthritis; Protease Inhibitors; Synovial Fluid; Trypsin Inhibitors

Topics: Arthritis, Rheumatoid; Cartilage; Chymotrypsin; Elastin; Electrophoresis, Polyacrylamide Gel; Granulocytes; Histones; Humans; Hyaluronoglucosaminidase; Hydrogen-Ion Concentration; Hydrolysis; Leukocytes; Lymphocytes; Peptide Hydrolases; Potassium Chloride; Proteoglycans; Synovial Fluid

Topics: Antithrombins; Arthritis, Rheumatoid; Chymotrypsin; Enzyme Inhibitors; Esterases; Humans; Plasma; Protease Inhibitors; Trypsin Inhibitors

Topics: Amino Acids; Amyloid; Arthritis, Rheumatoid; Carboxypeptidases; Chromatography, Gel; Chromatography, Ion Exchange; Chromatography, Paper; Chromatography, Thin Layer; Chymotrypsin; Cyanogen Bromide; Humans; Liver; Molecular Weight; Peptide Fragments; Spectrophotometry, Ultraviolet; Trypsin

Topics: Arthritis; Arthritis, Rheumatoid; Chymotrypsin; Cyclophosphamide; Femoral Artery; Heparin; Humans; Injections, Intra-Articular; Lupus Erythematosus, Systemic; Scleroderma, Systemic; Spondylitis, Ankylosing

Topics: Animals; Antibodies; Arthritis, Rheumatoid; Chymotrypsin; Haplorhini; Hemagglutination Inhibition Tests; Hemagglutination Tests; Humans; Immune Sera; Immunoglobulin G; Immunoglobulin M; Peptide Hydrolases; Trypsin; Ultracentrifugation

Topics: Arthritis, Rheumatoid; Blood; Chymotrypsin; Enzymes; Humans; In Vitro Techniques

Topics: Anti-Bacterial Agents; Arthritis; Arthritis, Rheumatoid; Cataract; Cataract Extraction; Chymotrypsin; Diabetes Mellitus; Drug Therapy; Endophthalmitis; Eye Diseases; Eye Foreign Bodies; Humans; Lens, Crystalline; Ophthalmia, Sympathetic; Panophthalmitis; Postoperative Complications; Steroids; Uveitis; Vitreous Body

Topics: Anti-Bacterial Agents; Arthritis; Arthritis, Infectious; Arthritis, Rheumatoid; Chymotrypsin; Humans; Trypsin

Topics: Arthritis; Arthritis, Rheumatoid; Chymotrypsin; Humans; Proteins