Strong crossreaction of human anti-aprotinin antibodies from heart transplant patient with [Arg15]aprotinin.
We detected anti-aprotinin antibodies by an enzyme immunoassay in serum of a 33 year old man who showed anaphylactic reactions during heart transplantation under aprotinin reexposition. The antibodies were isolated by affinity chromatography by aprotinin immobilized on CNBr activated Sepharose. The crossreactivity was tested by a competitive enzyme immunoassay (50% inhibition) against different aprotinin homologues and two human Kunitz-type protease inhibitors, bikunin and TFPI. In comparison with native aprotinin (immunoreactivity = 100%) the crossreaction of the homologue [Arg15]aprotinin was 76%, of [Val15]aprotinin 15% and of isoaprotinin 1, [Ala14,38]aprotinin and [seco15/16]aprotinin less than 10%. An immunoreactivity with bikunin and TFPI was not detected. Similar results were obtained with polyclonal anti-aprotinin antibodies from rabbit. Our results show that human anti-aprotinin antibodies are mainly directed against the reactive site of aprotinin. From this we conclude that the reactive site exposes the major epitope resulting in a major target site for antibodies in a species independent way, and therefore, it is obvious that the recombinant aprotinin homologue [Arg15]aprotinin, which is scheduled for therapy in open-heart surgery, will have similar immunogenic effects as native aprotinin.
Brinkmann
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Institut für Laboratoriums und Transfustonsmedizin, Her,- und Diabeteszentrum Nordrhein-Westfalen, Ruhr-Universität Bochum, Bad Oeynhausen, Germany. hdz.iltm@post.uni-bielefeld.de
Körfer
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Wenzel
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Tschesche
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Kleesiek
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D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D007611:Aprotinin / Q000276:immunology*; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D006023:Glycoproteins / Q000276:immunology / Q000494:pharmacology; D016027:Heart Transplantation / Q000276:immunology*; D006801:Humans; D007124:Immunoenzyme Techniques; D007074:Immunoglobulin G / Q000097:blood* / Q000302:isolation & purification; D008074:Lipoproteins / Q000276:immunology / Q000494:pharmacology; D008297:Male; D008562:Membrane Glycoproteins; D008969:Molecular Sequence Data; D011480:Protease Inhibitors / Q000276:immunology / Q000494:pharmacology; D011817:Rabbits; D017386:Sequence Homology, Amino Acid; D015842:Serine Proteinase Inhibitors / Q000276:immunology*; D014360:Trypsin Inhibitor, Kunitz Soybean
D006023:Glycoproteins; D007074:Immunoglobulin G; D008074:Lipoproteins; D008562:Membrane Glycoproteins; D011480:Protease Inhibitors; C109148:SPINT2 protein, human; D015842:Serine Proteinase Inhibitors; C051928:lipoprotein-associated coagulation inhibitor; C043017:trasylol, Arg(15)-; D007611:Aprotinin; D014360:Trypsin Inhibitor, Kunitz Soybean