Frank J D, Gould R J, Schaffer L W, Davidson J T, Gibson R E, Patrick D H, Vonderfecht S L, Cartwright M E
Department of Safety Assessment, Merck Sharp and Dohme Research Laboratories, West Point, PA 19486.
Histochemistry. 1992 May;97(4):355-60. doi: 10.1007/BF00270038.
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 snap-frozen. During infusion of echistatin (2.3 micrograms/kg/min), circulating platelet counts decreased from 331,000/mm3 to 167,000/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.
一种市售的抗人血小板糖蛋白IIIa小鼠单克隆抗体，被用于证明在静脉注射echistatin（一种源自锯鳞蝰蛇毒液的新型纤维蛋白原受体拮抗剂）后，从狒狒获取的肝活检组织中血小板的隔离情况。在给予echistatin之前，采集肝脏和脾脏的活检组织。肝活检组织要么在氟利昂 - 22/液氮中速冻，要么固定在10%中性缓冲福尔马林中。脾脏活检组织则速冻。在输注echistatin（2.3微克/千克/分钟）期间，循环血小板计数从331,000/mm³降至167,000/mm³。使用全身γ相机成像确认了肝脏内的选择性隔离，以证明在血小板减少发作期间，¹¹¹铟 - 奥克辛标记的血小板在肝脏内积聚。再次采集肝活检组织，要么在氟利昂 - 22/液氮中速冻，要么固定在10%中性缓冲福尔马林中。脾脏和腹股沟淋巴结的活检组织也速冻。作为阳性对照的富含血小板血浆涂片、脱蜡石蜡切片以及肝脏、脾脏和淋巴结的冰冻切片，采用抗生物素蛋白 - 生物素化过氧化物酶复合物（ABC）技术，用小鼠抗人血小板糖蛋白IIIa单克隆抗体进行染色。在输注echistatin之前，肝脏内的血小板染色极少。输注echistatin后，肝脏冰冻切片显示肝血窦内有明显血小板染色。淋巴结未显示定位，但脾脏在输注echistatin前后均显示明显血小板染色。富含血小板血浆涂片呈强阳性。在福尔马林固定、石蜡包埋的材料中未观察到明显血小板染色。因此，这种免疫细胞化学技术可能有助于在狒狒和其他灵长类动物的组织冰冻切片中定位血小板。
