Serum concentration of 19 serum proteins in Crohn's disease and ulcerative colitis.
(33/100)
The serum concentration of 19 serum proteins was determined by electrophoresis in 42 patients with Crohn's disease and 36 patients with ulcerative colitis. The results were compared with 78 healthy persons as matched controls. Distinctive, but similar, changes were present in the two diseases. An increased serum concentration of orosomucoid, alpha(1)-antitrypsin, easily precipitable glycoprotein, alpha(1)-antichymotrypsin, haptoglobin, and haemopexin was present. The serum concentration was decreased for prealbumin, albumin, alpha(2)-HS glycoprotein, caeruloplasmin, alpha(2)-macro-globulin, and transferrin. No significant difference between the two diseases existed as far as the serum protein pattern was concerned. Certain proteins, ;the acute phase reactants' (orosomucoid, alpha(1)-antitrypsin, alpha(1)-antichymotrypsin, and haptoglobin) and the immunoglobulins were clinically useful, since their serum concentration reflected the grade of activity of the disease. A pronounced elevation of haptoglobin compared with that of the other ;acute phase reactants' was present in patients with Crohn's disease complicated by suppurative fistulas or abscesses. Patients with active Crohn's disease who responded favourably to medical treatment had significantly higher immunoglobulin levels than patients not responding. A similar observation, though not statistically significant, was made in patients with ulcerative colitis. (+info)
The separation of alpha-2 macroglobulin into five components with differing electrophoretic and enzyme-binding properties.
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The alpha-2 macroglobulins from human serum and plasma were isolated by Bio-Gel P-300 and A5m gel filtration. The material showed a single peak on sedimentation velocity ultracentrifugation, a mol wt of 650,000 by sedimentation equilibrium ultracentrifugation, and a major precipitin arc in the alpha-2 macroglobulin region by immunoelectrophoresis against whole human serum. Two bands were observed in the alpha-2 macroglobulin region when acrylamide gel electrophoresis was performed with a pH 8.9 running gel. When a pH 7.8 gel was used, five electrophoretic species were observed. In both cases, the preaddition of stoichiometric amounts of trypsin or chymotrypsin added to alpha-2 macroglobulin resulted in disappearance of slower bands leaving only one band on acrylamide gel electrophoresis patterns. Preparative acrylamide gel electrophoresis separated alpha-2 macroglobulin obtained from Bio-Gel into five closely-spaced species. Separation was sufficiently adequate to show that those species of alpha-2 macroglobulin which bound trypsin and chymotrypsin were represented by slower moving species and that the fastest moving material had lost virtually all of the ability to bind these enzymes. Preparative acrylamide gel electrophoresis of a mixture of alpha-2 macroglobulin-trypsin complex and alpha-2 macroglobulin revealed that the fast moving component was alpha-2 macroglobulin-trypsin complex and that the slower moving material was unbound alpha-2 macroglobulin. The naturally occurring amidase activity of the alpha-2 macroglobulin using benzoylarginine-p-nitroanilide (BAPNA) as substrate was investigated and unlike its trypsin-binding activity, amidase activity was found to be of the same specific activity in all electrophoretic fractions. Binding of trypsin and chymotrypsin to alpha-2 macroglobulin revealed that alpha-2 macroglobulin maximally bound 2 moles of trypsin and 1 mole of chymotrypsin. When the enzymes were added simultaneously there was competition. Chymotrypsin added to alpha-2 macroglobulin before the addition of trypsin prevented all trypsin binding even though only one site was filled with chymotrypsin. These results were explained by the acrylamide gels which showed that 1 mole of chymotrypsin was sufficient to convert all the alpha-2 macroglobulin to a species with the fastest mobility which no longer binds additional enzyme. (+info)
Radioimmunoassay of CSF for encephalitogenic basic protein: a diagnostic test for MS?
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Competitive inhibition of binding between radioiodine-labelled encephalitogenic basic protein from human myelin ((125)I-HEProt) and normal human alpha-2 macroglobulin and between (125)I-HEProt and rabbit antiHEProt serum was used to study concentrated cerebrospinal fluid (CSF) under "blind" control for cross-reactivity with HEProt. Samples of CSF from patients meeting the standard criteria for definite MS and possible MS, and from patients with optic neuritis and "other" diagnoses were studied. CSF from patients in all four groups was shown to have an inhibitor cross-reactive with HEProt when studied by the (125)I-HEProt/alpha-2 macroglobulin test, but the amount was significantly greater in the definite MS group than in the "other" group. Results of the two tests on CSF from MS patients correlated, suggesting that the tests were identifying the same inhibitor. It was concluded that CSF contains an inhibitor similar to HEProt and that the amount present in CSF could be a useful diagnostic marker of MS. (+info)
A Waldenstrom macroglobulin that is both a cold agglutinin and a cryoglobulin because it binds N-acetylneuraminosyl residues.
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A purified human monoclonal IgM(kappa) (cold agglutinin MKV) has been characterized as both a cold agglutinin and a cryoglobulin. Since its reactivity with human erythrocytes but not with dog erythrocytes is reduced by treatment of the cells with ficin and its reactivity with both is abolished by treatment of the cells with neuraminidase, it has by definition Pr(2) specificity [Roelcke, D. (1974) Clin. Immunol. Immunopath. 2, 266-280]. Presumably, the membrane receptors for cold agglutinin MKV are sialic acid-containing glycoproteins in human cells and sialic acid-containing glycolipids in dog cells. Agglutination of erythrocytes is specifically inhibited by II(3)-N-acetylneuraminosyllactosylceramide (GM(3)) and N-acetylneuraminosylparagloboside but not by their N-glycolylneuraminosyl forms or by lactosylceramide or paragloboside, and the reactivity of neuraminidase-treated cells can be restored by allowing them to absorb either GM(3) or N-acetylneuraminosylparagloboside. When large amounts of ganglioside are absorbed, the cells are agglutinated not only in the cold, but also at 37 degrees , showing that the density of receptor sites on the erythrocyte surface can influence the thermal amplitude of cold agglutinins. Liberation of sialic acid from cold agglutinin MKV by treatment with neuraminidase (acylneuraminosyl hydrolase; EC 3.2.1.18) does not affect its agglutinating properties but the asialoprotein is no longer a cryoglobulin. Apparently the physical basis for its precipitation in the cold is the intermolecular immune binding of N-acetylneuraminosyl residues. (+info)
Plasma inhibitors of the components of the fibrinolytic pathway in man.
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The effect of highly purified inhibitor of the first component of complement (CaINH), alpha2 macroglobulin (alpha2M), and alpha1 antitrypsin on the components of the fibrinolytic pathway in human plasma has been examined. CaINH was the only factor active upon the Hageman factor fragments functioning at the initial step of the fibrinolytic pathway, alpha2M was the only factor active against the plasminogen activator and the most active inhibitor of plasmin. The inhibition of plasmin by alpha2M appeared stoichiometric with one molecule of alpha2M inhibiting two molecules of plasmin. All three plasma inhibitors were active against plasmin. (+info)
Physicochemical and biological properties of human and canine plasmins.
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Three kinds of plasmin were found to be generated when plasminogen or [(125)I]plasminogen was incubated at 32 degrees C for longer than 20 min in urokinase and 50% glycerol. Each plasmin was then separated by G-200 or G-75 Sephadex filtration, and its physicochemical properties were determined. The molecular weights of the three plasmins as determined by G-200 Sephadex filtration were 125,000+/-5,000(SD), 63,000+/-2,000(SD), and 31,500+/-1,000(SD), and those by sodium dodecyl sulfate(SDS)-polyacrylamide electrophoresis were 130,000+/-5,000(SD), 64,000+/-3,000(SD), and 32,000+/-1,500(SD). It was also found that during the incubation of the smallest plasmin in SDS and beta-mercaptoethanol it was further split into two smaller pieces of about 16,000 mol wt and that polymer proteins of 95,000+/-2,000(SD) and 48,000+/-1,500(SD) mol wt were formed. Despite these differences in the molecular size of the three plasmins, the specific activity of each plasmin was closely similar and in case of human plasmins averaged 29+/-0.9(SD) CTA units/mg plasmin and in case of canine plasmins 8.5+/-0.54(SD) CTA units/mg plasmin. Then, using human plasmin of the smallest size (mol wt 31,500), the total plasma antiplasmin capacity was determined in 20 normal human plasma, which averaged 7.8+/-2(SD) CTA units of plasmin per milliliter plasma. Studies were next made of the affinity of human [(125)I]-plasmin of the smallest size with albumin, gamma globulin, alpha(2)-macroglobulin, alpha(1)-antitrypsin, fibrinogen, and fibrin. The results were 0%, 0%, 14.6+/-0.5(SD)%, 17.6+/-0.6(SD)%, 21+/-0.5(SD)%, and 20.5+/-0.6(SD)%, respectively, of [(125)I]plasmin available and were unaltered when the amount of [(125)I]plasmin was increased to twice and four times the original amount. Finally, the plasma disappearance half-life of canine [(125)I]plasmin of the smallest size was studied in five healthy dogs, which averaged 14.2+/-0.63(SD) h. These results support the concept that the combination between plasmin and plasmin inhibitors is reversible and indicate that fibrinogen and fibrin have greater affinities than alpha(2)-macroglobulin or alpha(1)-antitrypsin. (+info)
An inherited X-linked serum system in man; the Xm system.
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The detection of an inherited X-linked serum system in man, disclosed by a heteroantisenim made specific by absorption, is described. These studies suggest that the antigen, demonstrated by the specific antiserum, resides in the alpha(2)-macroglobulin fraction of serum. The system has been named the Xm system, where X refers to the localization of the gene on the X chromosome, and the m indicates that the antigen is part of the alpha(2)-macroglobulin fraction. The distribution of phenotypes in unrelated individuals from 4 populations, as well as studies performed in families are consistent with an X-linked dominant mode of inheritance. One Xm(a-) daughter was found in a family where the father was Xm(a+). This finding is discussed with particular reference to the possible influence of sex and age on the development of the phenotype. This common X-linked marker is likely to prove useful in mapping the human X chromosome. (+info)
Nonabsorbable rabbit anti-Salmonella typhimurium antibody as detected by the complement-mediated bactericidal reaction.
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Herzberg, Mendel (University of Florida, Gainesville), Kathryn V. Kenny, and John B. Robbins. Nonabsorbable rabbit anti-Salmonella typhimurium antibody as detected by the complement-mediated bactericidal reaction. J. Bacteriol. 91:1548-1555. 1966.-A portion of antibody active in the complement-mediated bactericidal reaction against Salmonella typhimurium from hyperimmune rabbit serum has been shown to be nonabsorbable by repeated serial absorptions with whole heat-killed or living bacteria. The first two absorptions remove 90 to 95% of the activity, but 1 to 5% cannot be removed by subsequent absorptions. The nonabsorbable antibody appears to be a macroglobulin by density-gradient centrifugation and by comparison of activity and absorbability of purified gamma-M and gamma-G immune globulins. Alternative hypotheses involving low avidity antibody or antibody to minor cell antigenic components are offered in explanation of the phenomenon. (+info)