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  • 1
    ISSN: 1434-3932
    Keywords: Schlüsselwörter Biokompatibilität ; Antigenität ; Polyester ; Kollagen ; Key words Biocompatibility ; Antigenicity ; Polyester ; Collagen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Abstract At present there is neither an completely inert biomaterial available, nor does a universal test exist which objectively evaluates biocompatibility. One reason is the individuality of the host, especially with regard to the inflammatory response. Inflammation was found to induce biodegradation by hydrolysis or auto-oxidation of vascular prosthetic matrix after implantation. The present study was performed to investigate the specific humoral immune response after implantation of segments of a collagen-impregnated polyester prosthesis (Dacron) in Balb/c mice on experimental days 1, 18, 38, and 322. Serum antibody detection was performed by modified enzyme immunoassay using the prosthesis as a target. Specific polymer antibodies, enhanced by repeated implantations, could be detected in all mice which received grafts up to experimental day 322. These antibodies were not directed against the collagen coating. The antibody formation could be further enhanced by a combined administration of complete Freund's adjuvant together with the first implantation at experimental day 1. Results suggest that specific polymer antibodies, reflecting an inflammatory response, might be an additional parameter for biocompatibility testing of vascular prostheses.
    Notes: Zusammenfassung Bisher stehen keine komplett inerten Biomaterialien zur Verfügung und es existiert kein universeller Test zur Objektivierung der Biokompatibilität. Dies resultiert aus der individuellen Variabilität des Empfängerorganismus, insbesondere hinsichtlich der entzündlichen Reaktionsbereitschaft. Auch nach Implantation von Gefäßprothesen aus polymeren Biomaterialien kommt es zu einem chronischen Entzündungsprozeß. Dieser führt ursächlich durch Hydrolyse oder Autoxidation zur Biodegradation des Implantats. Mit unseren Untersuchungen galt es, eine möglicherweise bestehende, spezifische humorale Immunantwort nach Implantation von Segmenten einer kollagenimprägnierten Polyesterprothese (Dacron) in einem Tiermodell darzulegen. Balb/c-Mäusen wurde am 1., 18., 38. und 290. Versuchstag ein Prothesensegment intraperitoneal implantiert. Die Bestimmung der Serumantikörper erfolgte mit einem modifizierten Enzymimmunoassay unter Verwendung der Prothese als Target. Spezifische Antikörper gegen Polymere wurden nach wiederholter Implantation bei allen Tieren bis zum 322. Versuchstag nachgewiesen. Dabei konnte eine Antikörperbildung gegen die Kollagenimprägnierung ausgeschlossen werden. Die Antikörperbildung wurde durch den Zusatz von komplettem Freund-Adjuvans in Verbindung mit der ersten Implantation verstärkt. Der Nachweis von spezifischen Antikörpern gegen Polymere könnte zukünftig ein Parameter zur Testung der Biokompatibilität darstellen.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-5233
    Keywords: Key words Murine monoclonal glutamate decarboxylase antibodies ; Autoantibodies ; Type 1 diabetes mellitus ; Stiff-man syndrome
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To study the immune response to glutamic acid decarboxylase (GAD) in insulin-dependent diabetes mellitus, monoclonal GAD antibodies after fusion of splenocytes from a nondiabetes-susceptible BALB/c mouse immunized with human recombinant GAD65 were generated. Of the 44 monoclonals, 35 are specific for the GAD65 isoform, whereas 9 also react with GAD67. Some 37 monoclonals, including all GAD65/67 reactive antibodies, react with GAD by Western blot analysis. The remaining 7 GAD65 monoclonals bind GAD only in an immunoprecipitation assay, which implies that they target epitopes dependent on the conformation of the GAD molecule. The 125I-GAD binding of the GAD65 monoclonals reactive on Western blotting was significantly diminished by all 3 sera from Stiff-man syndrome patients but only by 3/30 (10%) sera from type 1 diabetic patients. In contrast, the 7 monoclonal antibodies reactive with a conformation-dependent GAD epitope were competitive with 83% of GAD-autoantibody-positive sera from these diabetic patients. Using chimeric GAD65/67 proteins, the epitope region targeted by these monoclonals was mapped to the middle of GAD65 (amino acids 221–442). This central conformation-dependent GAD region was also targeted by sera from patients with type 1 diabetes. In conclusion, our data show that even after common immunization of a nondiabetes-susceptible mouse strain, monoclonals were obtained which preferentially react with the GAD65 linear amino-terminus (amino acids 4–17) and a conformation-dependent region located in the middle of GAD targeted by autoantibodies, indicating that this GAD region is not restricted to the autoimmune response associated with the Stiff-man syndrome and the beta-cell destruction in type 1 diabetes mellitus.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1432-5233
    Keywords: Murine monoclonal glutamate decarboxylase antibodies ; Autoantibodies ; Type 1 diabetes mellitus ; Stiff-man syndrome
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract To study the immune response to glutamic acid decarboxylase (GAD) in insulin-dependent diabetes mellitus, monoclonal GAD antibodies after fusion of splenocytes from a nondiabetes-susceptible BALB/c mouse immunized with human recombinant GAD65 were generated. Of the 44 monoclonals, 35 are specific for the GAD65 isoform, whereas 9 also react with GAD67. Some 37 monoclonals, including all GAD65/67 reactive antibodies, react with GAD by Western blot analysis. The remaining 7 GAD65 monoclonals bind GAD only in an immunoprecipitation assay, which implies that they target epitopes dependent on the conformation of the GAD molecule. The125I-GAD binding of the GAD65 monoclonals reactive on Western blotting was significantly diminished by all 3 sera from Stiff-man syndrome patients but only by 3/30 (10%) sera from type 1 diabetic patients. In contrast, the 7 monoclonal antibodies reactive with a conformation-dependent GAD epitope were competitive with 83% of GAD-autoantibody-positive sera from these diabetic patients. Using chimeric GAD65/67 proteins, the epitope region targeted by these monoclonals was mapped to the middle of GAD65 (amino acids 221–442). This central conformation-dependent GAD region was also targeted by sera from patients with type 1 diabetes. In conclusion, our data show that evne after common immunization of a nondiabetes-susceptible mouse strain, monoclonals were obtained which preferentially react with the GAD65 linear amino-terminus (amino acids 4–17) and a conformation-dependent region located in the middle of GAD targeted by autoantibodies, indicating that this GAD region is not restricted to the autoimmune response associated with the Stiff-man syndrome and the bete-cell destruction in type 1 diabetes mellitus.
    Type of Medium: Electronic Resource
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