Rimoldi M T, Olabuenaga S E, de Bracco M M
J Protozool. 1981 Aug;28(3):351-4. doi: 10.1111/j.1550-7408.1981.tb02864.x.
Phagocytosis of culture forms of Trypanosoma cruzi was assayed by a radioisotopic method. Purified polymorphonuclear leukocytes (PMN) were mixed with 3H-uridine-labeled T. cruzi epimastigotes in the presence or absence of anti-T. cruzi antibodies. The reaction was stopped by adding N-ethyl-maleimide, and noningested parasites were lysed by complement. The percentage of radioactivity incorporated into the PMN pellet was recorded. The phagocytosis reaction was rapid, yielding maximum incorporation at 30 min at which point the radioactivity associated with the PMN cells decreased through release of the isotope to the supernatant. The degree of incorporation of radio-labeled parasites was a function of the effector/target cell ratio and the antibody concentration. The method is suitable for the quantitative determination of phagocytosis of T. cruzi by normal PMN.
通过放射性同位素方法测定了克氏锥虫培养形式的吞噬作用。在存在或不存在抗克氏锥虫抗体的情况下,将纯化的多形核白细胞(PMN)与3H-尿苷标记的克氏锥虫前鞭毛体混合。通过加入N-乙基马来酰亚胺终止反应,未摄取的寄生虫通过补体裂解。记录掺入PMN沉淀中的放射性百分比。吞噬反应迅速,在30分钟时达到最大掺入量,此时与PMN细胞相关的放射性通过同位素释放到上清液中而降低。放射性标记寄生虫的掺入程度是效应细胞/靶细胞比率和抗体浓度的函数。该方法适用于定量测定正常PMN对克氏锥虫的吞噬作用。