Abdi K, Kobzik L, Li X, Mentzer S J
Department of Surgery, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Lab Invest. 1995 Apr;72(4):445-52.
Carbohydrates play an important role in both the regulation and expression of endothelial cell surface molecules.
To investigate the molecular phenotype of endothelial cells in the sheep lung, we used a panel of 39 lectins and carefully titrated lectin histochemistry to identify small vessel glycoconjugates. Expression of glycoconjugates on isolated cells was studied by flow cytometry and lectin Western blotting.
Eight lectins stained small vessel endothelium with minimal background staining. To confirm cell surface binding, endothelial cells from the peripheral lung were isolated and tested for lectin recognition by flow cytometry. Three lectins (Datura stramonium, Griffonia simplicifolia-1, Lycopersicon esculentum) also stained isolated lung cells by flow cytometry. A lectin Western transfer technique demonstrated common binding to a high molecular weight (100 to 130 kD) band.
The use of lectins as probes of cell surface carbohydrate expression supports the possibility of selective glycoconjugate expression on sheep endothelium.
碳水化合物在调节和表达内皮细胞表面分子方面发挥着重要作用。
为了研究绵羊肺中内皮细胞的分子表型,我们使用了一组39种凝集素,并仔细滴定凝集素组织化学以鉴定小血管糖缀合物。通过流式细胞术和凝集素蛋白质印迹法研究了分离细胞上糖缀合物的表达。
八种凝集素对小血管内皮进行染色,背景染色极少。为了确认细胞表面结合,分离外周肺的内皮细胞,并通过流式细胞术测试凝集素识别情况。三种凝集素(曼陀罗、非洲豆蔻凝集素-1、番茄)也通过流式细胞术对分离的肺细胞进行染色。凝集素蛋白质印迹转移技术证明与高分子量(100至130 kD)条带存在共同结合。
使用凝集素作为细胞表面碳水化合物表达的探针支持绵羊内皮细胞上选择性糖缀合物表达的可能性。