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荧光偏振研究、脂质分析以及包裹在囊泡中的羧基荧光素摄取实验所显示的,不同组成的脂质体与兔胸腺细胞相互作用时兔胸腺细胞的特殊行为。

Peculiar behaviour of rabbit thymocytes in interaction with liposomes of different compositions shown by fluorescence polarization studies, lipid analysis, and uptake of vesicle-entrapped carboxyfluorescein.

作者信息

Roozemond R C, Urli D C

出版信息

Biochim Biophys Acta. 1982 Aug 12;689(3):499-512. doi: 10.1016/0005-2736(82)90308-x.

Abstract

In order to obtain more information on membrane phenomena occurring at the cell surface of rabbit thymocytes we have performed experiments aimed at altering the lipid composition of the plasma membrane. Thymocytes were incubated at 37 degree C with phospholipid vesicles of different compositions. Vesicle-cell interaction was followed by measuring the degree of fluorescence polarization and the uptake of vesicle-entrapped carboxyfluorescein. Neutral and negatively charged liposomes prepared from egg phosphatidylcholine are currently used in investigations of vesicle-cell interaction. In this report we show that these liposomes do not interact with rabbit thymocytes as is evident from unaltered lipid fluidity measured in whole cells and in isolated plasma membranes. This was confirmed by experiments with vesicle-entrapped carboxyfluorescein showing hardly any uptake of the fluorophor from neutral and negatively charged egg phosphatidylcholine liposomes. Using both techniques substantial interaction was found with positively charged egg phosphatidylcholine liposomes and with liposomes prepared from soybean lecithin which is composed of a variety of phospholipids. The results of these experiments were supported by lipid analysis of cells treated with soybean lecithin liposomes. Increase in phosphatidylcholine contents of mixed phospholipid vesicles was further shown to result in decreased vesicle-cell interaction. From measurements of the quantity of carboxyfluorescein inside cells and the total amount of cell-associated carboxyfluorescein it is concluded that adsorption plays a prominent role in interaction between liposomes and rabbit lymphocytes. The grade of maturation of lymphocytes was also found to affect vesicle-cell interaction. The more mature thymocytes took up more vesicle-entrapped carboxyfluorescein from soybean liposomes than immature thymocytes. Mesenteric lymph node cells exhibited a still stronger interaction. The role of vesicle and cell surface charge and membrane fluidity of both vesicles and cells in interaction between liposomes and rabbit thymocytes is discussed.

摘要

为了获取更多关于兔胸腺细胞表面发生的膜现象的信息,我们进行了旨在改变质膜脂质组成的实验。胸腺细胞在37℃下与不同组成的磷脂囊泡一起孵育。通过测量荧光偏振程度和囊泡包裹的羧基荧光素的摄取来跟踪囊泡与细胞的相互作用。由鸡蛋磷脂酰胆碱制备的中性和带负电荷的脂质体目前用于囊泡与细胞相互作用的研究。在本报告中,我们表明这些脂质体不与兔胸腺细胞相互作用,这从全细胞和分离的质膜中测量的未改变的脂质流动性中可以明显看出。用囊泡包裹的羧基荧光素进行的实验证实了这一点,该实验表明几乎没有从中性和带负电荷的鸡蛋磷脂酰胆碱脂质体中摄取荧光团。使用这两种技术,发现与带正电荷的鸡蛋磷脂酰胆碱脂质体以及由多种磷脂组成的大豆卵磷脂制备的脂质体有大量相互作用。用大豆卵磷脂脂质体处理的细胞的脂质分析支持了这些实验的结果。进一步表明,混合磷脂囊泡中磷脂酰胆碱含量的增加会导致囊泡与细胞的相互作用减少。从细胞内羧基荧光素的量和细胞相关羧基荧光素的总量的测量结果可以得出结论,吸附在脂质体与兔淋巴细胞之间的相互作用中起主要作用。还发现淋巴细胞的成熟程度会影响囊泡与细胞的相互作用。与未成熟的胸腺细胞相比,成熟度更高的胸腺细胞从大豆脂质体中摄取的囊泡包裹的羧基荧光素更多。肠系膜淋巴结细胞表现出更强的相互作用。讨论了囊泡和细胞表面电荷以及囊泡和细胞的膜流动性在脂质体与兔胸腺细胞相互作用中的作用。

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