Summary
A commercially available mouse monoclonal antibody to human platelet glycoprotein IIIa was used to demonstrate sequestration of platelets in hepatic biopsies obtained from baboons following intravenous infusion of echistatin, a novel fibrinogen receptor antagonist derived from the venom of the snake Echis carinatus. Biopsies of liver and spleen were taken prior to administration of echistatin. The hepatic biopsies were either snap-frozen in Freon-22/liquid nitrogen or fixed in 10% neutral buffered formalin. Biopsies of spleen were snapfrozen. During infusion of echistatin (2.3 µg/kg/min), circulating platelet counts decreased from 331000/mm3 to 167000/mm3. Selective sequestration within the liver was confirmed using whole body gamma camera imaging to demonstrate 111Indium-oxine labeled platelet accumulation within the liver during the thrombocytopenic episode. Hepatic biopsies were again taken and either snap-frozen in Freon-22/liquid nitrogen or fixed in 10% neutral buffered formalin. Biopsies of spleen and inguinal lymph node were also snap-frozen. Platelet rich plasma smears, included as positive controls, dewaxed paraffin sections, and cryosections of liver, spleen, and lymph node were stained with monoclonal mouse anti-human platelet glycoprotein IIIa using an avidin biotinylated peroxidase complex (ABC) technique. Prior to infusion of echistatin, platelet staining within the liver was minimal. After echistatin infusion, hepatic cryosections showed prominent platelet staining within hepatic sinusoids. No localization was shown in lymph node, however, the spleen showed prominent platelet staining both before and after echistatin infusion. Platelet rich plasma smears were intensely positive. No prominent platelet staining was observed in formalin-fixed, paraffin-embedded material. Thus, this immunocytochemical technique may help localize platelets in cryosections of tissues from baboons and other primate species.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Boerboom LE, Olinger GN, Tie-Zhu L, Rodriguez ER, Ferrans VJ, Kissebah AH (1990) Histologic, morphometric, and biochemical evolution of vein bypass grafts in a nonhuman primate model. I. Sequential changes within the first three months. J Thorac Cardiovasc Surg 99:97–106
Cipolle RJ, Rodvold KA, Seifert R, Clarens R, Ramirez-Lassepas M (1983) Heparin-associated thrombocytopenia: a prospective evaluation of 211 patients. Ther Drug Monit 5:205–211
Coller BS, Folts JD, Scudder LE, Smith SR (1986) Antithrombotic effect of a monoclonal antibody to the platelet glycoprotein IIb/IIIa receptor in an experimental animal model. Blood 68:783–786
Dewanjee MK, Rao SA, Didisheim P (1981) Indium-111 tropolone, a new high affinity platelet label: preparation and evaluation of labeling parameters. J Nucl Med 22:981–987
Drouin J, Izaguirre CA, Patenaude P (1988) Quantitation of cell membrane glycoproteins in pathological conditions using a lectin-bound enzyme-linked immunosorbent assay (ELISA). J Immunol Methods 110:217–223
Eldrup-Jorgensen J, Connolly RJ, Mackey WC, Ramberg K, O'Donnell TF, Valeri CR, Callow AD (1986) Antiplatelet therapy and vascular grafts. Studies in a baboon ex vivo shunt. Arch Surg 121:778–781
Francis RB (1989) Clinical disorders of fibrinolysis: a critical review. Blut 59:1–14
Gaetano G de, Garattini S (eds) (1978) Platelets: a multidisciplinary approach. Raven Press, New York, pp 65, 293, 303
Gan ZR, Gould RJ, Jacobs JW, Friedman PA, Polokoff MA (1988) Echistatin. A potent platelet aggregation inhibitor from the venom of the viper, Echis carinatus. J Biol Chem 263:19827–19832
Garsky VM, Lumma PK, Freidinger RM, Pitzenberger SM, Randall WC, Veber DF, Gould RJ, Friedman PA (1989) Chemical synthesis of echistatin, a potent inhibitor of platelet aggregation from Echis carinatus: synthesis and biological activity of selected analogs. Proc Natl Acad Sci USA 86:4022–4026
Hampton JW, Matthews C (1966) Similarities between baboon and human blood clotting. J Appl Physiol 21:1713–1716
Hanson SR, Harker LA (1988) Interruption of acute platelet-dependent thrombosis by the synthetic antithrombin d-phenylalanyl-l-prolyl-l-arginyl chloromethyl ketone. Proc Natl Acad Sci USA 85:3184–3188
Hanson SR, Harker LA, Bjornsson TD (1985a) Effects of plateletmodifiying drugs on arterial thromboembolism in baboons. Aspirin potentiates the antithrombotic actions of dipyridamole and sulfinpyrazone by mechanism(s) independent of platelet cyclooxygenase inhibition. J Clin Invest 75:1591–1599
Hanson SR, Kotze HF, Savage B, Harker LA (1985b) Platelet interactions with Dacron vascular grafts. A model of acute thrombosis in baboons. Arteriosclerosis 5:595–603
Hanson SR, Pareti FI, Ruggeri ZM, Marzec UM, Kunicki TJ, Montgomery RR, Zimmerman TS, Harker LA (1988) Effects of monoclonal antibodies against the platelet glycoprotein IIb/IIIa complex on thrombosis and hemostasis in the baboon. J Clin Invest 81:149–158
Heyns AduP, Lotter MG, Badenhorst PN, Kotze H, Killian FC, Herbst C, Reenen OR van, Minnaar PC (1980) Kinetics and in vivo redistribution of 111Indium-labelled human platelets after intravenous protamine sulphate. Thromb-Haemost 44:65–68
Kapsch DN, Adelstein EH, Rhodes GR, Silver D (1979) Heparin-induced thrombocytopenia, thrombosis, and hemorrhage. Surgery 86:148–155
Kapsch DN, Silver D (1981) Heparin-induced thrombocytopenia with thrombosis and hemorrhage. Arch Surgery 116:1423–1427
King DJ, Kelton JG (1984) Heparin-associated thrombocytopenia. Ann Intern Med 100:535–540
Koerker DJ, Harker LA, Goodner CJ (1975) Effects of somatostatin on hemostasis in baboons. N Engl J Med 293:476–479
Mackey WC, Connolly RJ, Callow AD, Keough EM, Ramberg-Laskaris K, McCullough JL, O'Donnell TF, Melaragno A, Valeri CR, Weiblen B (1984) Aspirin decreases platelet uptake on Dacron vascular grafts in baboons. Ann Surg 200:93–99
Marshall LR, Herrmann RP (1989) Australian snake venoms and their in vitro effect on human platelets. Thrombos Res 54:269–275
McManus LM, Pinckard RN, Fitzpatrick FA, O'Rourke RA, Crawford MH, Hanahan DJ (1981) Acetyl glyceryl ether phosphorylcholine. Intravascular alterations following intravenous infusion into the baboon. Lab Invest 45:303–307
Murphy WG, Kelton JG (1988) Idiosyncratic drug-induced thrombocytopenia. Curr Stud Hematol Blood Transf 54:71–88
Rehman A, Hayee A, Nagi AH (1984) Effects of prolonged poisoning by Russell's viper venom on blood coagulation, platelets, and fibrinolysis. Jpn J Med Sci Biol 37:1–7
Robertson DN, Page CP (1987) Effect of platelet agonists on airway reactivity and intrathoracic platelet accumulation. Br J Pharmacol 92:105–111
Todd ME, McDevitt E, Goldsmith EI (1972) Blood-clotting mechanisms of nonhuman primates. Choice of the baboon model to simulate man. J Med Prim 1:132–141
Vargaftig BB, Lefort J, Prancan AV, Chignard M, Benveniste J (1979) Platelet-lung in vivo interactions: an artifact of a multipurpose model? Haemostasis 8:171–182
Warkentin TE, Kelton JG (1989) Heparin-induced thrombocytopenia. Ann Rev Med 40:31–44
Wood GS, Warnke R (1981) Suppression of endogenous avidinbinding activity in tissues and its relevance to biotin-avidin detection systems. J Histochem Cytochem 29:1196–1204
Wordinger RJ, Miller GW, Nicodemus DS (1983) Manual of immunoperoxidase techniques. American Society of Clinical Pathologists Press, Chicago, p 35
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Frank, J.D., Gould, R.J., Schaffer, L.W. et al. Immunocytochemical localization of platelets in baboon hepatic sinusoids using monoclonal mouse anti-human platelet glycoprotein IIIa following induction of thrombocytopenia. Histochemistry 97, 355–360 (1992). https://doi.org/10.1007/BF00270038
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00270038