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Developmental research on the application of a microbial lectin CLA for the analysis of human-specific erythrocyte surface antigen.

微生物凝集素 CLA 在人类特异性红细胞表面抗原分析中的应用进展研究。

基本信息

批准号：
60860010
负责人：
OISHI Kunio
金额：
$ 7.04万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Developmental Scientific Research
财政年份：
1985
资助国家：
日本
起止时间：
1985 至 1986
项目状态：
已结题

项目摘要

Possibility of practical use of CLA, a chitooligosaccharide-binding lectin from a fungus Conidiobolus lamprauges, was investigated. CLA binded to human erythrocytes highly specifically. It agglutinated human erythrocytes nonspecifically for blood groups at a concentration of 25 <micro> g protein/ml, agglutinated the erythrocytes of chimpanzee, gibbons, and tufted capuchin at an order of 1,000 <micro> g/ml, but agglutinated none of the erythrocytes of 47 species of other mammals, birds, reptiles, amphibia, and fishes. Animal species-specificity of CLA was too strict to apply CLA for the comparative studies on animals in taxonomy, evolutionology, and mass genetics, but suitable for human-animal blood diagnosis in police science. CLA detected human blood stains as old as 10 years and as little as 2.5 <micro> l. Differing from known anti-blood group antibodies, CLA was not inhibited by saliva and semen. This property gave an great advantage to CLA when it was used as the reagent for the mass diagnosis of occult blood in the early checkup of cancer on digestive organs. Generally CLA did not bind to human hemopoietic cells. Only exception was HEL, human erythroleukemia, cells. Tetradecanoyl phorbor acetate treatment of HEL cells resulted in the loss of CLA-bindability. CLA will be applicable for the clinical diagnosis of inherent and acquired blood diseases and the studies on the action of differentiation-inhibitory and stimulatory substances. CLA receptor on human erythrocytes was not glycolipid but glycoprotein, and in glycoprotein, not glycophorin and spectrin but Band 3 glycoprotein. Chemical and enzymological studies showed that the CLA required-for binding to the receptor the presence of L-fucosyl residue and a special structure in more than 5 sugar residues on nonreducing terminal of lactosaminoglycan chain of Band 3.

本文研究了从一种真菌耳霉（Conidiobolus lamprauges）中提取的一种结合壳寡糖的凝集素--共轭亚油酸（CLA）的实际应用的可能性。CLA与人红细胞高度特异性结合。在25 g蛋白/ml的浓度下，它非特异性地凝集人类红细胞<micro>的血型，凝集黑猩猩、长臂猿和簇绒卷尾猴的红细胞的数量级为1，000 <micro>g/ml，但不凝集其他47种哺乳动物、鸟类、爬行动物、两栖动物和鱼类的红细胞。CLA的动物种属特异性太强，不能用于动物的分类学、进化学和群体遗传学的比较研究，但适用于公安科学中的人畜血液诊断。CLA检测到的人类血迹年龄最大为10岁，最小为2.5<micro>升。与已知的抗血型抗体不同，CLA不受唾液和精液的抑制。这一性质使CLA在消化道肿瘤早期检查中作为隐血的集体诊断试剂具有很大的优势。CLA一般不与人造血细胞结合。唯一的例外是HEL，人红白血病细胞。十四酰佛波酯处理HEL细胞导致CLA结合性丧失。共轭亚油酸可用于先天性和后天性血液病的临床诊断以及分化抑制物质和分化刺激物质作用的研究。人红细胞上CLA受体不是糖脂而是糖蛋白，糖蛋白中不是血型糖蛋白和血影蛋白而是带3糖蛋白。化学和酶学研究表明，CLA与受体结合需要L-岩藻糖残基的存在和带3乳糖胺聚糖链非还原末端5个以上糖残基的特殊结构。