Halbreich A, Roger J, Pons J N, Geldwerth D, Da Silva M F, Roudier M, Bacri J C
Laboratoire des Milieux Désordonnés et Hétérogènes, Université Pierre-et-Marie-Curie, Paris, France.
Biochimie. 1998 May-Jun;80(5-6):379-90. doi: 10.1016/s0300-9084(00)80006-1.
The use of cell-targeted ferrofluid in the characterization of modifications of cell membranes is reviewed. Maghemite ferrofluid was synthesized by the Massart method, complexed with dimercaptosuccinic acid (FF). Cell targeting by FF was developed by coupling FF to various biological effectors such as antibodies, lectins, etc, which enabled magnetic cell sorting. Modifications in erythrocyte membranes were studied using FF bound to recombinant human annexin V (AnxFF) which is very sensitive, compared to other Anx-based reagents, in the early detection of phosphatidylserine (PS) exposition on the outer leaflet of the plasma membrane. Thus PS exposition on mouse RBC was detected already after a 24-h storage at 4 degrees C and, transiently, 24 h after their infection by Plasmodium parasites, at which time the parasites are still confined to the liver, thus leading to the recruitment of young RBC and the accumulation of a species, intermediate between reticulocytes and erythrocytes, and the actual RBC target of plasmodial invasion. AnxFF revealed PS exposition on RBC from sickle cell anemia patients, following various inflammations and already after 20 days of human blood storage under blood bank conditions. Such a sensitive detection should be similar to that of macrophages which recognize exposed PS on cells and bring about the latter's elimination from the circulation. AnxFF binding determination was combined with that of cell electrophoretic mobility, glycerol resistance and filterability to characterize RBC membrane modifications in Alzheimer's disease patients which suggested a continuous damage and regeneration in RBC of these patients. A logistic analysis suggested that several three-parameter combinations could permit diagnosis of Alzheimer's disease with up to 95% accuracy. THP1 cells and macrophages, derived themselves by incubation with retinoic acid, were bound to FF and placed in a radio frequency alternating magnetic field. Magnetocytolysis was associated with FF attachment to the cells without damage to non-bound cells and without heating of the surrounding solution.
本文综述了细胞靶向铁磁流体在细胞膜修饰表征中的应用。采用马萨特法合成了磁赤铁矿铁磁流体，并与二巯基琥珀酸（FF）络合。通过将FF与各种生物效应物（如抗体、凝集素等）偶联，开发了FF的细胞靶向技术，实现了磁性细胞分选。利用与重组人膜联蛋白V结合的FF（AnxFF）研究红细胞膜的修饰，与其他基于膜联蛋白的试剂相比，AnxFF在早期检测质膜外小叶上磷脂酰丝氨酸（PS）暴露方面非常敏感。因此，在4℃储存24小时后以及感染疟原虫24小时后（此时寄生虫仍局限于肝脏），小鼠红细胞上的PS暴露已被检测到，这导致了年轻红细胞的募集以及一种介于网织红细胞和红细胞之间的细胞的积累，而这种细胞正是疟原虫入侵的实际红细胞靶点。AnxFF揭示了镰状细胞贫血患者红细胞在各种炎症后以及在血库条件下储存20天后PS的暴露情况。这种灵敏的检测应该类似于巨噬细胞对细胞上暴露的PS的识别，并导致后者从循环中清除。将AnxFF结合测定与细胞电泳迁移率、甘油抗性和过滤性测定相结合，以表征阿尔茨海默病患者红细胞膜的修饰，这表明这些患者的红细胞存在持续的损伤和再生。逻辑分析表明，几种三参数组合可实现高达95%准确率的阿尔茨海默病诊断。THP1细胞和通过与视黄酸孵育获得的巨噬细胞与FF结合，并置于射频交变磁场中。磁细胞溶解与FF附着于细胞相关，对未结合的细胞无损伤，且周围溶液不发热。
