Abstract
We have employed the method of Burwen and Satir (J. Cell Biol., 1977, 74:690) to measure the disappearance of surface folds from resident guinea pig peritoneal macrophages after antibody-dependent phagocytosis. Unilamellar phospholipid vesicles containing dimyristoylphosphatidylcholine and 1 mol % dinitrophenyl-epsilon- aminocaproyl-phosphatidylethanolamine, a lipid that possesses a hapten headgroup, were prepared by an ether injection technique. These vesicles were taken up by macrophages in a time- and temperature- dependent fashion. Vesicles that contained ferritin trapped in the internal aqueous volume were identified within macrophages by transmission electron microscopy. Scanning electron microscopy has shown that macrophage surface folds decrease dramatically after phagocytosis. The surface fold length (micrometer) per unit smooth sphere surface area (micrometer2) decreases from 1.3 +/- 0.3 micrometer- 1 to 0.53 +/- 0.25 micrometer-1 when cells are incubated in the presence of specific anti-DNP antibody and vesicles at 37 degrees C. No significant effect was observed in the presence of antibody only or vesicles only. Our studies shown that phagocytosis is associated with a loss of cell surface folds and a loss of cell surface area, which is consonant with current views of the endocytic process. On the basis of our uptake data, we estimate that approximately 400 micrometer2 of vesicle surface membrane is internalized. The guinea pig macrophage plasma membrane has a total area of approximately 400 micrometer2 in control studies, whereas the cells have roughly 300 micrometer2 after phagocytosis. These estimates of surface areas include membrane ruffles and changes directly related to changes in cell volume. We suggest that during antibody-dependent phagocytosis a membrane reservoir is made available to the cell surface.
Full Text
The Full Text of this article is available as a PDF (854.9 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Burwen S. J., Satir B. H. Plasma membrane folds on the mast cell surface and their relationship to secretory activity. J Cell Biol. 1977 Sep;74(3):690–697. doi: 10.1083/jcb.74.3.690. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bøyum A. Isolation of lymphocytes, granulocytes and macrophages. Scand J Immunol. 1976 Jun;Suppl 5:9–15. [PubMed] [Google Scholar]
- Daems W. T., Brederoo P. Electron microscopical studies on the structure, phagocytic properties, and peroxidatic activity of resident and exudate peritoneal macrophages in the guinea pig. Z Zellforsch Mikrosk Anat. 1973 Nov 5;144(2):247–297. doi: 10.1007/BF00307305. [DOI] [PubMed] [Google Scholar]
- Deamer D., Bangham A. D. Large volume liposomes by an ether vaporization method. Biochim Biophys Acta. 1976 Sep 7;443(3):629–634. doi: 10.1016/0005-2736(76)90483-1. [DOI] [PubMed] [Google Scholar]
- Edelson P. J., Zwiebel R., Cohn Z. A. The pinocytic rate of activated macrophages. J Exp Med. 1975 Nov 1;142(5):1150–1164. doi: 10.1084/jem.142.5.1150. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Esser A. F., Bartholomew R. M., Parce J. W., McConnell H. M. The physical state of membrane lipids modulates the activation of the first component of complement. J Biol Chem. 1979 Mar 25;254(6):1768–1770. [PubMed] [Google Scholar]
- Fedorko M. E., Cross N. L., Hirsch J. G. Appearance and distribution of ferritin in mouse peritoneal macrophages in vitro after uptake of heterologous erythrocytes. J Cell Biol. 1973 May;57(2):289–305. doi: 10.1083/jcb.57.2.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kaplan J. Evidence for reutilization of surface receptors for alpha-macroglobulin.protease complexes in rabbit alveolar macrophages. Cell. 1980 Jan;19(1):197–205. doi: 10.1016/0092-8674(80)90401-8. [DOI] [PubMed] [Google Scholar]
- Lewis J. T., Hafeman D. G., McConnell H. M. Kinetics of antibody-dependent binding of haptenated phospholipid vesicles to a macrophage-related cell line. Biochemistry. 1980 Nov 11;19(23):5376–5386. doi: 10.1021/bi00564a036. [DOI] [PubMed] [Google Scholar]
- Muller W. A., Steinman R. M., Cohn Z. A. The membrane proteins of the vacuolar system. II. Bidirectional flow between secondary lysosomes and plasma membrane. J Cell Biol. 1980 Jul;86(1):304–314. doi: 10.1083/jcb.86.1.304. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Petty H. R., Hafeman D. G., McConnell H. M. Specific antibody-dependent phagocytosis of lipid vesicles by RAW264 macrophages results in the loss of cell surface Fc but not C3b receptor activity. J Immunol. 1980 Dec;125(6):2391–2396. [PubMed] [Google Scholar]
- Petty H. R. Response of the resident macrophage to concanavalin A. Alterations of surface morphology and anionic site distribution. Exp Cell Res. 1980 Aug;128(2):439–454. doi: 10.1016/0014-4827(80)90079-8. [DOI] [PubMed] [Google Scholar]
- Schwartz M. A., McConnell H. M. Surface areas of lipid membranes. Biochemistry. 1978 Mar 7;17(5):837–840. doi: 10.1021/bi00598a014. [DOI] [PubMed] [Google Scholar]
- Stahl P., Schlesinger P. H., Sigardson E., Rodman J. S., Lee Y. C. Receptor-mediated pinocytosis of mannose glycoconjugates by macrophages: characterization and evidence for receptor recycling. Cell. 1980 Jan;19(1):207–215. doi: 10.1016/0092-8674(80)90402-x. [DOI] [PubMed] [Google Scholar]
- Steinman R. M., Brodie S. E., Cohn Z. A. Membrane flow during pinocytosis. A stereologic analysis. J Cell Biol. 1976 Mar;68(3):665–687. doi: 10.1083/jcb.68.3.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Walters M. N., Papadimitriou J. M. Phagocytosis: a review. CRC Crit Rev Toxicol. 1978 Sep;5(4):377–421. doi: 10.3109/10408447809081012. [DOI] [PubMed] [Google Scholar]