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1.
Microheterogeneity forms of alpha 1-acid glycoprotein as indicators of rheumatoid arthritis activity 总被引:6,自引:0,他引:6
A Mackiewicz T Paw?owski A Mackiewicz-Paw?owska K Wiktorowicz S Mackiewicz 《Clinica chimica acta; international journal of clinical chemistry》1987,163(2):185-190
The microheterogeneity of alpha 1-acid glycoprotein (AGP) has been studied in the sera of 48 patients with rheumatoid arthritis and of 12 healthy individuals. For each rheumatoid patient the disease activity has been assessed and each patient has been assigned to one of four activity grades: I, inactive; II, mildly active; III, moderately active; and IV, severe. Concanavalin A-affinity electrophoresis revealed three microheterogeneity variants of AGP: non-reactive with Con A, weakly reactive with Con A and strongly reactive with Con A. The relative amounts of AGP-variants observed in the healthy donors were similar to those observed in the patients with activity grade I, but differed significantly from patients with grades II, III and IV. The differences between the AGP-patterns of patients with activity grades II, III and IV were also statistically significant. 相似文献
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M Jezequel N S Seta M M Corbic J M Feger G M Durand 《Clinica chimica acta; international journal of clinical chemistry》1988,176(1):49-57
In order to test whether abnormalities in hepatocytes affect the glycoprotein carbohydrate moiety, crossed immunoaffinoelectrophoresis (CIAE) with Concanavalin A (Con A) was used to study serum alpha 1-acid glycoprotein (alpha 1-AGP) and alpha 2-HS glycoprotein (alpha 2-HS) obtained from alcoholic patients with biopsy-proven liver disease. Cirrhotic patients, placed in groups C1, C2 or C3, according to Pugh's classification, were compared to healthy donors (N) and to steatosic non-cirrhotic patients (S). Con A CIAE patterns revealed in group N three subpopulations for alpha 2-HS and four for alpha 1-AGP. Two main results emerged from this study: (1) in the alcoholic groups, the proportions of Con A-unreactive subpopulations of both glycoproteins increased. Moreover, group N could be separated from group S and group S from all the cirrhotic groups. (2) There was a good correlation between the relative amounts in Con A-unreactive subpopulations of alpha 1-AGP and alpha 2-HS. The increases observed in Con A-unreactive subpopulations are probably a general phenomenon related to alterations in glycosylation processing during liver cell damage. 相似文献
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D Biou P Chanton D Konan N Seta H N'Guyen J Feger G Durand 《Clinica chimica acta; international journal of clinical chemistry》1989,186(1):59-66
To determine whether alterations of the carbohydrate moiety of human alpha 1-acid glycoprotein constitute a marker of hepatic damage we studied purified alpha 1-acid glycoprotein from healthy individuals and two groups of patients with benign liver diseases: alcoholic cirrhosis and acute viral hepatitis. The results indicate: (1) increased concanavalin A-non reactive forms in cirrhosis and hepatitis, (2) a markedly increased proportion of fucosyl residues in all cirrhotic and some hepatitis patients. Although hyperfucosylation is generally considered to be a tumor marker, the observation here in the two benign liver diseases indicates that an increased fucosyl content should be considered as a more general expression of pathological glycoconjugate metabolism. 相似文献
6.
Protection by alpha 1-acid glycoprotein against tumor necrosis factor- induced lethality 总被引:3,自引:0,他引:3
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《The Journal of experimental medicine》1994,180(4):1571-1575
We here report that alpha 1-acid glycoprotein, a typical acute phase protein, protects mice from lethal shock induced by tumor necrosis factor (TNF) or endotoxin. The protection is observed both in normal and in galactosamine-sensitized mice. Optimal desensitization requires at least 3 mg alpha 1-acid glycoprotein administered 2 h before the lethal challenge. Under these conditions, complete inhibition of all TNF-induced metabolic changes was observed: fall in body temperature, release of liver transaminases, enhanced clotting time, and mortality. The known platelet aggregation-inhibitory activity of alpha 1-acid glycoprotein provides a possible explanation for this protective capacity. 相似文献
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The molecular properties of alpha 1-acid glycoprotein are briefly discussed. This molecule has been shown in in vitro experiments to have both a stabilizing effect on vascular permeability and antiinflammatory properties. We were able to demonstrate these two effects in vivo in guinea pigs (skin, Evan's Blue extravasation) and in rats (paw, carrageenan induced inflammation). Further experiments were performed in rats relating to possible therapeutic indications for alpha 1-acid glycoprotein: (1) inhibitory effect on brain edema formation after experimental stroke, (2) therapeutic effect in the puromycin aminonucleoside-induced minimal change nephrosis, (3) improvement of vital parameters in hemorrhagic-hypovolemic shock, (4) increase in survival rate in septic peritonitis, and (5) promising effects in burn-induced remote lung injury. The high content of sialic acid and the high negative charge of alpha 1-acid glycoprotein are believed to be major contributors to its stabilizing effect on vascular permeability. The protein is bound to the glycocalyx of the endothelial cells (and presumably to structures of the glomerular basement membrane), thereby hindering the passage of other polyanionic molecules through the vascular wall. The antiinflammatory/immunomodulatory effect of alpha 1-acid glycoprotein appears mainly due to suppression of polymorphonuclear neutrophils. This action is dependent on the glycan part of the molecule, which is highly variable (microheterogeneity). It is obvious that there are differences between the different glycan forms as far as the antiinflammatory property of the protein is concerned. Together with data in the literature, the results presented here suggest a variety of potential indications for therapeutic use of alpha 1-acid glycoprotein in humans. 相似文献
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This is a noncompetitive enzyme-linked immunosorbent assay for measuring low concentrations (2 to 100 micrograms/L) of human alpha 1-acid glycoprotein (AGP; orosomucoid). The method is based on a simple "sandwich" technique involving polyclonal rabbit antisera against AGP. Mean within-run and total (between-run) CVs were 6.2% and 9.7%, respectively. Analytical recovery, tested in various biological fluids, averaged 101%. The technique has been successfully applied to diluted biological fluids such as bronchoalveolar lavage, cerebrospinal and amniotic fluids, and hepatocyte-culture supernates. Because of its analytical validity and the commercial availability of the reagents, this assay is suitable for large-scale determinations of AGP concentrations in those biological fluids in which its concentration is relatively low. 相似文献
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There is an increasing demand for quantification of serum alpha 1-acid glycoprotein (AAG, orosomucoid) in studies evaluating the protein binding of highly bound basic drugs. This paper describes an adaptation of an automated immunoturbidimetric assay for this protein to the Cobas Bio centrifugal analyzer. Replicate analyses of aliquots from six different solutions were used in determining precision. We also analyzed 367 patients' serum samples, in duplicate, to determine the distribution of AAG in hospitalized patients. The intra- and inter-run CVs ranged from 1.3% to 4.4% and from 0.6% to 6.6%, respectively. AAG concentrations in patients' samples ranged from 0.38 to 3.16 g/L. Results by this method correlate well with those by radial immunodiffusion, with no significant amount of bias between the two methods. This immunoturbidimetric procedure is faster and less expensive than currently used radial immunodiffusion techniques, and precision is acceptable. 相似文献
10.
A "sandwich"-type enzyme-linked immunosorbent assay for determining concentrations of human alpha 1-acid glycoprotein (AGP) is described. Microtiter plates coated with a polyclonal rabbit antibody to human AGP were subsequently incubated with the antigen, with a specific murine monoclonal antibody, and with goat anti-mouse immunoglobulins conjugated to alkaline phosphatase. To evaluate the method for assay of AGP in human sera, we compared it with single radial immunodiffusion and "rocket" electroimmunoassay. The respective correlations were r = 0.988 (n = 45) and r = 0.973 (n = 47). Repeated assays of a human serum sample with an average AGP concentration of 859 mg/L yielded within-day and between-day CVs of 1.4% (n = 5) and 6.3% (n = 10), respectively. Because of its low detection limit (4.4 micrograms/L), this assay is also suitable for determination of AGP concentrations in other biological fluids, such as dialysates of patients being treated by continuous ambulatory peritoneal dialysis. 相似文献
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D Biou C Bauvy H N'Guyen P Codogno G Durand M Aubery 《Clinica chimica acta; international journal of clinical chemistry》1991,204(1-3):1-12
The carbohydrate moiety of purified alpha 1-acid glycoprotein (AGP) from healthy male adults (AGPn) and late-term pregnant women (AGPp) was analysed. Polyacrylamide gel electrophoresis with sodium dodecyl sulfate before and after N-glycanase treatment showed that AGPp had a slightly higher molecular mass due to an enriched carbohydrate moiety. BIO-GEL P-4 and Concanavalin A (Con A)-Sepharose chromatography of the oligosaccharides released by hydrazinolysis and fractionated by high-voltage electrophoresis indicated a progression towards Con A-unbound oligosaccharides and towards larger glycans in pregnancy. Carbohydrate analysis of purified AGPp and AGPn and of the most increased oligosaccharide fraction (F4A) evidenced a decrease in the fucosyl molar ratio and a slight increase in the galactosyl, N-acetyl-glucosaminyl and N-acetyl neuraminyl ratios. These results suggest that AGP contains more highly branched oligosaccharides and/or additional N-acetyllactosamine-type oligosaccharides in pregnancy. 相似文献
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Sensitive chemiluminescence immunoassay for the determination of rat serum alpha1-acid glycoprotein.
We present the establishment of a sensitive immunoassay for the determination of alpha1-acid glycoprotein (orosomucoid, AGP) in rat serum. The assay is based upon antigen capture by surface-immobilized specific polyclonal rabbit anti-AGP antibodies with biotinylated rat AGP (rAGP) as the tracer, and formatted as competitive enzyme immunoassay. Signaling is performed by streptavidin-conjugated horseradish peroxidase. Enzyme activity is quantified by an enhanced chemiluminometric method, allowing the sensitive detection of rAGP serum levels in small sample volumes. 相似文献
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Y Ochi Y Fujiyama S Hosoda T Miyazaki M Yoshimura T Hachiya Y Kajita 《Clinica chimica acta; international journal of clinical chemistry》1982,122(2):145-160
We have demonstrated that 125 I-CEA (carcinoembryonic antigen) from two commercial sources (Roche and CIS RIA kit) can be precipitated by antibody to alpha 1-acid glycoprotein (AG) dose-dependently. The binding of 125 I-CEA to anti-AG can be displaced by unlabelled AG, though, there was no cross-reaction between AG and anti-CEA. The data strongly suggest that CEA has an immunological similarity to AG. The perchloric acid extract from cancerous tissue was fractionated on a Sephadex G-200 column, and the first eluted fraction (containing large amounts of CEA) was subjected to affinity chromatography using anti-AG bound to Sepharose. The bound fraction was eluted, labelled with 125 I, and then applied to a Sephadex G-200 column with carrier protein. The radioactivity was found mainly in the large molecular size eluted fraction (Fr 1). Almost all radioactivity of Fr 1 was precipitated by anti-AG and anti-CEA. This experiment also demonstrated that a large molecular weight component (Mr: 180 000) obtained from tumor was immunologically related to both CEA and AG. Based on differences in binding affinities between CEA with anti-CEA and with anti-AG we postulate that CEA may be a large molecular precursor of AG (big form of AG). 相似文献
15.
《The Journal of experimental medicine》1978,148(2):507-521
Specific antibodies against human alpha1-acid glycoprotein reacted with human lymphocytes, granulocytes, and monocytes. The antigen on the leukocytes is an externally located integral membrane glycoprotein which is made by the cells and has an apparent mol wt of 52,000. It is released from cells in vitro to the culture medium. The mol wt of the soluble fragment is 41,000, which corresponds to that of alpha1-acid glycoprotein in serum and urine. Peptide mapping confirmed that the main part of the cellular membrane antigen consists of alpha1-acid glycoprotein with an additional, probably hydrophobic fragment. This finding may partially explain the increase in the serum levels of alpha1-acid glycoprotein observed in many disorders involving leukocyte proliferation. In addition, the known sequence homology of alpha1-acid glycoprotein with immunoglobulins can now be more easily understood by their origin in similar cell types. 相似文献
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N Seta B Tissot F Forestier J Feger F Daffos G Durand 《Clinica chimica acta; international journal of clinical chemistry》1991,203(2-3):167-175
alpha 1-Acid glycoprotein concentrations and reactivity to concanavalin A were measured in maternal and fetal serum and amniotic fluid obtained from 24 women undergoing diagnostic cordocentesis at 20 to 33 wk gestation and in 30 additional fetal sera (19 to 34 weeks gestation). Maternal alpha 1-acid glycoprotein serum levels were five to ten times higher than fetal and amniotic levels. Fetal alpha 1-acid glycoprotein levels were found to increase with advancing gestational age. Using crossed immunoaffino electrophoresis with concanavalin A, alpha 1-acid glycoprotein patterns were identical in maternal serum and amniotic fluid but totally different in fetal serum. The fetal concanavalin A pattern changed progressively during fetal life towards that of the newborn. These data confirm earlier assumptions of fetal synthesis of alpha 1-acid glycoprotein and provide normal reference values for alpha 1-acid glycoprotein in fetal serum. In addition, the specific fetal concanavalin A pattern indicates that the alpha 1-acid glycoprotein glycosylation process during fetal life differs from that in post-natal life. 相似文献
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Electroimmunodiffusion methods of Laurell and radial immunodiffusion method of Mancini are compared for the qualitative and quantitative analysis of native and desialylated alpha 1-acid glycoprotein. Samples are incubated under different conditions at decreasing pH (3.5 to 0.5 pH units), with increasing ionic strength and with neuraminidase during different time intervals. Results show a pronounced decrease in electrophoretic mobility of alpha 1-acid glycoprotein treated either with acidic reagents or with neuraminidase (ionic strength has no effect). Such a procedure might involve chemical or enzymatic hydrolysis by which sialyl residues are removed. This hydrolysis implicates lower results in the estimation of the desialylated glycoprotein by electroimmunodiffusion. On the other hand, the amounts of alpha 1-acid glycoprotein evaluated by radial immunodiffusion are not modified after incubation. This is expected since diffusion and antigenic properties are not related to the sialic acid content. The data suggest that radial immunodiffusion, less accurate and sensitive than electroimmunodiffusion, is nevertheless more adequate for estimating native and desialylated alpha 1-acid glycoprotein. 相似文献
19.
Higai K Azuma Y Aoki Y Matsumoto K 《Clinica chimica acta; international journal of clinical chemistry》2003,329(1-2):117-125
BACKGROUND: In certain pathophysiological conditions, such as inflammation rheumatoid arthritis and diabetes mellitus (DM), alterations in asparagine-linked glycan (N-glycan) patterns of the acute-phase protein, alpha(1)-acid glycoprotein (AGP), have been reported. In this study, we investigated N-glycan structures of AGP purified from the sera of patients with acute inflammation (n=5), type 2 diabetes mellitus (n=5), and healthy individuals (n=5). METHODS: N-Glycans were released with peptide N-glycosidase F (PNGase F) from denatured AGP and purified with cellulose cartridge. N-glycans were analyzed by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOFMS) in combination with exoglycosidase digestion. RESULTS: We revealed increases in bi-antennary complex glycans and in alpha1-3 fucosylated bi-, tri-, and tetra-antennary glycans and a decrease in tri-antennary glycans in inflammation patients. These results support increases in bindings to concanavalin A (ConA) and Aleuria aurantia lectins (AALs). In diabetic patients, the pathogenesis-specific change in N-glycan patterns of AGP was not significant. CONCLUSIONS: The MALDI-TOFMS method is sensitive and suitable for profiling analysis of N-glycans in clinical samples. 相似文献
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We describe a solid-phase enzyme-linked immunosorbent assay for alpha 1-acid glycoprotein in human plasma. Plasma samples are incubated with alkaline phosphatase-linked, purified alpha 1-acid glycoprotein in alpha 1-acid glycoprotein-specific antibody-coated polystyrene tubes. The alkaline phosphatase that becomes attached to the tube via an immunological reaction between the alpha 1-acid glycoprotein and the specific antibody is measured spectrophotometrically. This assay is accurate reproducible, simple, and economical. As little as 4 microgram of alpha 1-acid glycoprotein per liter can be detected. The normal range for alpha 1-acid glycoprotein in the plasma of healthy adults, as measured by this method, is 0.48-1.27 g/L; the range is significantly different, 0.29-0.73 g/L, for women who are taking oral contraceptive pills. 相似文献