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.
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.
D000327:Adsorption; D000818:Animals; D002462:Cell Membrane / Q000187:drug effects; D002470:Cell Survival; D004563:Electrochemistry; D005452:Fluoresceins / Q000378:metabolism; D005454:Fluorescence Polarization; D008055:Lipids / Q000494:pharmacology; D008081:Liposomes / Q000378:metabolism / Q000494:pharmacology*; D008560:Membrane Fluidity / Q000187:drug effects; D010713:Phosphatidylcholines / Q000494:pharmacology; D011817:Rabbits; D013025:Glycine max; D013601:T-Lymphocytes / Q000187:drug effects / Q000502:physiology*
D005452:Fluoresceins; D008055:Lipids; D008081:Liposomes; D010713:Phosphatidylcholines; C024098:6-carboxyfluorescein