- Email: [email protected]
Reverse fibrin slide technique: A method for localization of plasminogen activator inhibitor activity in tissue sections
108
THROMBOSIS RESEARCH
Suppl. VI,
1986
212
PATTERNOF FIBRINOLYTICPARAMETERSIN PATIENTSWITH GASTROINTESTINAL CARCINOMAS J.C.Kirchheimer,K.Huber, 0. Wagner, B.R. Binder. Lab. Clin.-Exp.Physiol.,Universityof Vienna, A-1090 Vienna,Austria Changes in the plasma levels of componentsof the fibrinolyticsystem have been investigated in80 patients suffering from gastrointestinalcarcinomas. Urokinese antigen (RIA),tissue-typeplasminogenactivator antigen (ELISA)and plasminogen activator inhibitor (functional assay) were determined. In case of pancreatic and colorectalcarcinoma patients with metastases as well as those without metastases revealed significantly increasedplasma urokinaselevels.Patientssufferingfrom gall bladder or gastric carcinoma did not show significantly elevatedurokinaseantigen levels both as compared to agematched controls.Determinationof tissuetype plasminogen activator antigen in all four carcinoma groups did not reveal significant differences when compared to an age-matched healthy control group. The concentrationsof plasminogenactivator inhibitorwere significantlyincreasedin all carcinomagroups, exhibitingno differences between the patient groups with or without metastases. No correlations between the differentparameters of the fibrinolyticsystem could be obtained.
213
REVERSE FIBRIN SLIDE TECHNIQUE: A METHOD FOR LOCALIZATIONOF PLASMINCGEN ACTIVATOR INHIBITOR ACTIVITY IN TISSUE SECTIONS J. C. Kirchheimer, 0. Wagner, G. Christ, C.Korninger, B.R.Binder Lab. Clin.-Exptl.Physiol.,Univ. Vienna, A-1090 Vienna, Austria A reverse fibrin slide technique was developed to detect plasminogen activator inhibitorin tissue sections.Thereby a plasminogen-richfibrin film containingexogenouslyadded urokinase is placed on top of a tissue section, preincubated with antibodies against tissue-type plasminogen activatorfor 30 minutes.The incorporatedurokinasecauses a diffuse lysis within 60 minutes.The parts of the tissue section containingplasminogen activatorinhibitorcauses opaque lysis resistantzones in the fibrin film which disappearonly after prolongedincubation.When tissue sectionsfrom rat aorta and rat vena cava were studied, lysis-resistant zones were found in both cases predominantlyabove the endothelialcells and smooth muscle cells. Thus, this technique offers a new approach to localize the plasminogen activator inhibitor in tissue sections. Furthermore, using specific antibodies against the fast acting endothelieltype plasminogen activator inhibitor, a good correlation was found between activity localizationby the reversefibrin slide techniqueand antigen localization using immunofluorescence staining.
THROMBOSIS RESEARCH
Suppl. VI,
1986
212
PATTERNOF FIBRINOLYTICPARAMETERSIN PATIENTSWITH GASTROINTESTINAL CARCINOMAS J.C.Kirchheimer,K.Huber, 0. Wagner, B.R. Binder. Lab. Clin.-Exp.Physiol.,Universityof Vienna, A-1090 Vienna,Austria Changes in the plasma levels of componentsof the fibrinolyticsystem have been investigated in80 patients suffering from gastrointestinalcarcinomas. Urokinese antigen (RIA),tissue-typeplasminogenactivator antigen (ELISA)and plasminogen activator inhibitor (functional assay) were determined. In case of pancreatic and colorectalcarcinoma patients with metastases as well as those without metastases revealed significantly increasedplasma urokinaselevels.Patientssufferingfrom gall bladder or gastric carcinoma did not show significantly elevatedurokinaseantigen levels both as compared to agematched controls.Determinationof tissuetype plasminogen activator antigen in all four carcinoma groups did not reveal significant differences when compared to an age-matched healthy control group. The concentrationsof plasminogenactivator inhibitorwere significantlyincreasedin all carcinomagroups, exhibitingno differences between the patient groups with or without metastases. No correlations between the differentparameters of the fibrinolyticsystem could be obtained.
213
REVERSE FIBRIN SLIDE TECHNIQUE: A METHOD FOR LOCALIZATIONOF PLASMINCGEN ACTIVATOR INHIBITOR ACTIVITY IN TISSUE SECTIONS J. C. Kirchheimer, 0. Wagner, G. Christ, C.Korninger, B.R.Binder Lab. Clin.-Exptl.Physiol.,Univ. Vienna, A-1090 Vienna, Austria A reverse fibrin slide technique was developed to detect plasminogen activator inhibitorin tissue sections.Thereby a plasminogen-richfibrin film containingexogenouslyadded urokinase is placed on top of a tissue section, preincubated with antibodies against tissue-type plasminogen activatorfor 30 minutes.The incorporatedurokinasecauses a diffuse lysis within 60 minutes.The parts of the tissue section containingplasminogen activatorinhibitorcauses opaque lysis resistantzones in the fibrin film which disappearonly after prolongedincubation.When tissue sectionsfrom rat aorta and rat vena cava were studied, lysis-resistant zones were found in both cases predominantlyabove the endothelialcells and smooth muscle cells. Thus, this technique offers a new approach to localize the plasminogen activator inhibitor in tissue sections. Furthermore, using specific antibodies against the fast acting endothelieltype plasminogen activator inhibitor, a good correlation was found between activity localizationby the reversefibrin slide techniqueand antigen localization using immunofluorescence staining.