SULM – Schweizerische Union für Labormedizin | Union Suisse de Médecine de Laboratoire | Swiss Union of Laboratory Medicine

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K Miszti-Blasius 1 , J Kappelmayer 1 , L Xia 2 , RP McEver 2

1Department of Clinical Biochemistry and Molecular Pathology, University Medical School, Debrecen, Hungary, 2Oklahoma Medical Research Foundation, Oklahoma City, USA

Objective: During the association of leukocytes with platelets and endothelial cells, P-selectin glycoprotein ligand-1 (PSGL-1) binds to P-selectin expressed on activated platelets and endothelial cells. Our aim was to evaluate the response to various thrombotic stimuli in wild type and PSGL-1 knockout mice.
Methods: Mice were challenged by tail vein injection of (i) 15 ug collagen plus 2 ug epinephrine (coll/epi) (ii) thromboplastin (5x diluted Innovin) or (iii) saline. Retro-orbital samples were drawn to ACD anticoagulant and platelet and leukocyte counts were measured and plasmas were frozen for thrombin-antithrombin III (TAT) complex. In addition, kidneys, liver, spleen and lungs were investigated for fibrin deposition by immunohistochemistry.
Results: After coll/epi and thromboplastin challenge, mice of both genotypes decreased significantly platelet and leukocyte number. The degree of thrombocytopenia was milder after thromboplastin challenge, while the mean TAT complex values were 10-fold elevated in this group compared to saline injected animals (244 mg/L versus 23 mg/L). TAT values after thrombolpastin challenge were also significantly elevated compared the coll/epi treated amimals, but the two genotypes showed no significant difference. The major difference could be observed by investigating the lung and liver sections for fibrin deposition by immunohistochemistry. After coll/epi challenge the lungs of the PSGL-1 knockout animals contained less thrombi and in the liver no central vein involvement could be observed.
Summary and conclusions: Thromboplastin challenge produces more thrombin generation while coll/epi challenge results in a more pronounced thrombocytopenia. For the applied doses, the deficiency of PSGL-1 defended for neither effects, however the difference in the fine structure of the formed fibrin refers to the importance of this molecule in multicellular interaction during thrombus formation.


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