Cytochemical study of liposome and lipid vesicle phagocytosis

doi:10.1016/0005-2736(83)90262-6

Biochimica et Biophysica Acta (BBA) - Biomembranes

Volume 735, Issue 1, 26 October 1983, Pages 77-85

This paper is dedicated to Norman Davidson on the occasion of his 65th birthday.

https://doi.org/10.1016/0005-2736(83)90262-6 Get rights and content

Abstract

Electron microscopy cytochemistry has been used to study the cytoplasmic location of liposomes and lipid vesicles following specific antibody-dependent phagocytosis. The vesicle compositions were 94–99 mol% ‘fluid’ lipid (egg phosphatidylcholine or dimyristoylphosphatidylcholine at 37°C or ‘solid’ lipid (dipalmitoylphosphatidylcholine at 37°C). In some cases, 4 mol% phosphatidylserine was included in the vesicle membrane so as to vary the surface charge density. These vesicles undergo specific antibody-dependent phagocytosis by RAW264 macrophages when the lipid membranes contain 1–2 mol% dinitrophenyl lipid hapten in the presence of rabbit anti-dinitrophenyl IgG antibody. Internalized lipid vesicles can be visualized with the electron microscope when ferritin is trapped in the internal aqueous compartments prior to internalization. The lipid vesicles were demonstrated to be internal to the macrophage plasma membranes by selectively staining the plasma membranes with Ruthenium red. The cytoplasmic location of vesicles and liposomes was studied by electron microscopic staining for activities of the following enzymes: (1) acid phosphatase; (2) inorganic trimetaphosphatase; (3) adenosine triphosphatase; and (4) glucose-6-phosphatase. The first two enzymatic activities were found in association with ferritin-containing vesicles after antibody-dependent phagocytosis, showing the formation of vesicle-containing phagolysosomes. Adenosine triphosphatase and glucose-6-phosphatase were primary not associated with the vesicles, suggesting a minimal association of vesicles with plasma membrane, Golgi, endoplasmic reticulum and perinuclear cisternae. Phagosome-lysosome fusion did not appear to depend on the type of target lipid vesicle or liposome, on the ‘fluidity’ of the target membrane, or the presence of phosphatidylserine in the target membrane.

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Cytochemical study of liposome and lipid vesicle phagocytosis

Abstract

Biochim. Biophys. Acta

Biochim. Biophys. Acta

J. Ultrastruct. Res.

J. Ultrastruct. Res.

Exp. Cell Res.

Annu. Rev. Biochem.

J. Exp. Med.

Cancer Res.

Infect. Immun.

J. Immunol.

Biochemistry

Biochemistry

J. Cell Biol.

J. Cell Biol.