The Interaction of Cationic Liposomes Containing Entrapped Horseradish Peroxidase With Cells in Culture

Magee, Wayne E.; Goff, C. W.; Schoknecht, Jean D.; Smith, Mary D.; Cherian, Kay

doi:10.1083/jcb.63.2.492

Cited by 121 publications

(28 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…While this paper was in preparation, a study on the interaction of cationic liposomes containing horseradish peroxidase with cells in culture was reported by Magee et al (6). They reported that "liposomes containing horseradish peroxidase were adsorbed by HeLa cells at least 300 times as effectively as was free horseradish peroxidase".…”

Section: I17mentioning

confidence: 99%

Liposomes containing chelating agents. Cellular penetration and a possible mechanism of metal removal.

Rahman¹,

Wright²

1975

The Journal of Cell Biology

105

View full text Add to dashboard Cite

Electron microscope studies were done on mouse liver, from 5 min to 8 wk after an intravenous injection of liposomes containing ethylenediaminetetraacetic acid (EDTA). Livers of mice receiving an injection of liposomes containing KCl instead of EDTA or an injection of a solution of EDTA were also examined. Liposomes were shown to be phagocytized by hepatocytes as well as by Kupffer cells within minutes after the injection. Initially, there was a close contact between the liposomai membrane and the cellular membrane, followed by an invagination of the latter and the formation of a distinct vesicle surrounding a single liposome or a cluster of several liposomes. No fusion between the liposomal membrane and the cell membrane was observed. Between 15 min and 6 h after liposome injection, the Kupffer cells were found to have an increased number of lysosomes and autophagic vacuoles. Within the latter, morphologically intact liposomes or remnants of liposomes could be seen. At 12 h after injection, a striking increase in macrophages was observed in the liver sinusoids of EDTA-liposome-injected mice, but not in those of KCl-liposome-injected mice. Within the macrophages, remnants of liposomes occasionally could be observed. However, the origin and the physiological role of these cells are unknown. In the hepatocytes, morphological changes were first observed 24 h after injection; there were large numbers of autophagic vacuoles, and some cells showed extensive areas of focal cytoplasmic degeneration. The morphology of the liver cells returned to normal about 7 days after injection. No morphological changes were observed in livers of mice receiving EDTA solution without liposomes, A possible mechanism by which the liposome-encapsulated chelating agents can successfully remove intracellular toxic metals is discussed. The use of liposomes as carriers seems to be a useful tool for intracellular delivery of chelating agents or drugs in general.For the last 20 yr, the polyaminopolycarboxylic acid-chelating agents, ethylenediaminetetraacetic acid (EDTA) and diethylenetriaminepentaacetic acid (DTPA), have been the drugs of choice for therapy of poisoning by various metals. However, the inability of these chelating agents to cross cellular membranes (1, 3) seriously limits their efficacy in removing the intracellular fraction of 112

show abstract

Section: I17mentioning

confidence: 99%

Liposomes containing chelating agents. Cellular penetration and a possible mechanism of metal removal.

Rahman¹,

Wright²

1975

The Journal of Cell Biology

105

View full text Add to dashboard Cite

show abstract

“…Recent work in this (1)(2)(3) and other laboratories (4)(5)(6)(7) has shown that mammalian cells cultured in vitro can incorporate large numbers of lipid vesicles, so-called liposomes, without significant cytotoxicity. Since a variety of water-soluble materials can be trapped inside vesicles (3)(4)(5)(6)(7)(8) and various glycolipids and glycoproteins inserted into the vesicle wall (3,8), uptake of vesicles of defined composition by cells offers a potential method for modifying cellular composition and for introducing nonpermeable, biologically active molecules into cells.…”

mentioning

confidence: 99%

“…Since a variety of water-soluble materials can be trapped inside vesicles (3)(4)(5)(6)(7)(8) and various glycolipids and glycoproteins inserted into the vesicle wall (3,8), uptake of vesicles of defined composition by cells offers a potential method for modifying cellular composition and for introducing nonpermeable, biologically active molecules into cells. The use of vesicles as "carriers" to enhance incorporation of cyclic nucleotides (1,2), enzymes (3,6), sugars (3,4), and anti-tumor drugs (3,4) into cultured cells has already been demonstrated.…”

mentioning

confidence: 99%

Lipid vesicles as carriers for introducing materials into cultured cells: influence of vesicle lipid composition on mechanism(s) of vesicle incorporation into cells.

Poste¹,

Papahadjopoulos²

1976

Proc. Natl. Acad. Sci. U.S.A.

178

View full text Add to dashboard Cite

The mechanisms involved in the uptakt-of uni-and multi-lamellar lipid vesicles by BALB Recent work in this (1-3) and other laboratories (4-7) has shown that mammalian cells cultured in vitro can incorporate large numbers of lipid vesicles, so-called liposomes, without significant cytotoxicity. Since a variety of water-soluble materials can be trapped inside vesicles (3-8) and various glycolipids and glycoproteins inserted into the vesicle wall (3, 8), uptake of vesicles of defined composition by cells offers a potential method for modifying cellular composition and for introducing nonpermeable, biologically active molecules into cells. The use of vesicles as "carriers" to enhance incorporation of cyclic nucleotides (1, 2), enzymes (3, 6), sugars (3, 4), and anti-tumor drugs (3, 4) into cultured cells has already been demonstrated. In this communication we present data that suggest that vesicles enter cells both by fusion with the plasma membrane and by endocytosis. Our results indicate that the physical state of lipids in the vesicle is of major importance in determining which of these two uptake pathways predominates. Lipids and Preparation of Vesicles. Cholesterol, palmitic, and stearic acids were purchased from Fluka A. G. (Buchs, Switzerland). Phosphatidylcholine (PC) and phosphatidylserine (PS) were purified from egg yolk and bovine brain, respectively, as described previously (9). Distearylphosphatidylcholine (DSPC) and DPPC were synthesized as reported elsewhere (9). All lipids were finally purified on silicic acid columns, shown to be pure by thin layer chromatography, and stored (approximately 10 jimol/ml) in sealed ampoules under nitrogen at -50' until use. MATERIALS AND METHODS[3H]DPPC (specific activity 4 Ci/mmol) was prepared from dipalmitoyleylphosphatidylcholine by catalytic hydrogenation (10) and was mixed with the lipids in chloroform (1, 2). Multilamellar vesicles (MLV) were prepared by the method of Bangham et al. (11), and small unilamellar vesicles (SUV) by sonication of MLV preparations (12). MLV and SUV were dispersed in phosphate-buffered saline without Ca2+ and Mg2 , pH 7.4, at a concentration of 2 ,gmol/ml. Lipid concentrations were determined by phosphate analysis (10). Fluid vesicles with a net negative surface charge (hereafter called fluid charged vesicles) were prepared from a mixture of PS and PC (1:9 mol). Neutral fluid vesicles were prepared from PC alone. Lipids for both classes of vesicles were dispersed in phosphate-buffered saline and sonicated (SUV only) at 250. Differential scanning calorimetry (10) revealed that the vesicles had a gel-to-liquid crystalline transition temperature (TJ) (mid-point) at -10°. Negatively charged solid vesicles ("solid charged") were prepared from PS, DPPC, and DSPC (1:4.5:4.5 mol) and suspended and sonicated (SUV only) at 50°. Calorimetry revealed the T, (midpoint) to be 430 for SUV and 470 for MLV (Fig. 1). These vesicles are therefore "fluid" (above TJ) during preparation at 500. During incubation with cells at 370 the majority of lip...

show abstract

“…Liposomes are also under intensive investigation as vehicles for introducing membrane-impermeant substances (e.g., drugs, enzymes, chelating agents) into cells (2)(3)(4). Less attention has been devoted to the kinetics of vesicle-cell interaction, although a number of attempts have been made to define the mechanisms involved (5)(6)(7)(8)(9)(10).…”

mentioning

confidence: 99%

Liposome--lymphocyte interaction: saturable sites for transfer and intracellular release of liposome contents.

Blumenthal¹,

Weinstein²,

Sharrow³

et al. 1977

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

The water-soluble dye 6-carboxyfluorescein was trapped in the internal aqueous compartments of small sonicated dioleoyl lecithin vesicles and used to assess the kinetics of transfer of vesicle contents to human lymphocytes. By using flow microfluorometry, the initial rate of dye transfer to the cells was measured as a function of the concentration of vesicles in the external medium. plex. Liposome-cell interaction is currently being studied in a number of laboratories as a model for the fusion of biological membranes occurring in such processes as phagocytosis, secretion, cell division, and heterokaryon formation (1). The use of liposomest makes it possible to substitute a synthetic membrane of known and controllable composition for one of the natural ones (2, 3). Liposomes are also under intensive investigation as vehicles for introducing membrane-impermeant substances (e.g., drugs, enzymes, chelating agents) into cells (2-4). Less attention has been devoted to the kinetics of vesicle-cell interaction, although a number of attempts have been made to define the mechanisms involved (5-10).Liposomes can become associated with cells by stable adsorption to the cell surface, by endocytosis, or by fusion with the cell membrane (5, 6). Studies in our laboratory (11) and that of Huang et al. (12) indicate that neither stable adsorption nor metabolically active endocytosis is significant in the interaction of lymphocytes with vesicles whose lipids are in the fluid phase. Fusion and, perhaps, a form of endocytosis insensitive to glycolytic and respiratory inhibitors are therefore left as the major possible mechanisms for transfer of the contents of such vesicles to lymphocytes. The combination of lymphocytes and fluid phase vesicles thus provides a relatively simple system for a detailed study of the kinetics of the transfer process.In collaboration with Hagins and Yoshikami we recently developed a sensitive technique for monitoring the transfer of a trapped marker from liposomes into cells (11) using the highly water-soluble fluorescent dye 6-carboxyfluorescein (6-CF). In those studies, incubation of human peripheral blood lymphocytes with small unilamellar lipid vesicles containing a high concentration of 6-CF resulted in a widespread distribution of fluorescence throughout the cytoplasm of each cell. "Selfquenching" largely prevented the dye from fluorescing as long as it remained trapped at high concentration in vesicles; relief of self-quenching as the dye was released into the much larger volume of a cell enabled us to monitor transfer of 6-CF from vesicle to lymphocyte. To avoid the mechanistic implications of terms such as "fusion," we use the operational term "transfer" to indicate passage of 6-CF, with accompanying dilution, into the cell interior.In the present study, we measured the initial rate of 6-CF transfer from fluid phase unilamellar lipid vesicles to lymphocytes as a function of liposome concentration under various conditions. Kinetic analysis revealed both a nonsaturable and a saturable compo...

show abstract

The Interaction of Cationic Liposomes Containing Entrapped Horseradish Peroxidase With Cells in Culture

Cited by 121 publications

References 24 publications

Liposomes containing chelating agents. Cellular penetration and a possible mechanism of metal removal.

Liposomes containing chelating agents. Cellular penetration and a possible mechanism of metal removal.

Lipid vesicles as carriers for introducing materials into cultured cells: influence of vesicle lipid composition on mechanism(s) of vesicle incorporation into cells.

Liposome--lymphocyte interaction: saturable sites for transfer and intracellular release of liposome contents.

Contact Info

Product

Resources

About