Abstract
The influence of several metabolic inhibitors and pharmacologic agents on macrophage deformation (induced by fluid shear stress) was examined in relationship to changes in ATP content and phagocytosis of latex beads. Two relatively specific inhibitors of glycolysis (iodoacetate [IA], and sodium fluoride [NaF]) and a sulfhydryl-binding agent (N-ethylmaleimide [NEM] markedly inhibited phagocytosis and reduced cell deformability. A microtubule-disrupting agent (vinblastine) and a highly specific inhibitor of glycolysis (2-deoxyglucose) markedly inhibited phagocytosis without influencing cell deformability. An organomercurial sulfhydryl binding agent p-chloromercuribenzene (PCMBS) and a microfilament-disrupting agent (cytochalasin B) inhibited phagocytosis and increased cell deformability. The effects of these agents on phagocytosis and cell deformability bore no consistent relationship to alterations in cellular content of ATP. The observation that 2-deoxyglucose, the most specific inhibitor of glycolysis examined, reduced ATP content to levels far lower (15 percent of control values) than those achieved by any other agent examined and inhibited phagocytosis without altering cell deformability, suggests that alterations in cell deformability induced by NaF, IA, NEM, PCMBS, and cytochalasin B are not due to inhibition of glycolysis per se, but instead result from direct or indirect effects of these agents on cell constituents, possibly contractile proteins, which are determinants of cell deformability. The finding that cytochalasin B, NEM, PCMBS, and IA interfere with phagocytosis and alter cell deformability, together with evidence that these agents interact with isolated actin and myosin, suggests that contractile proteins are important both in phagocytosis and as determinants of cell deformability. The observation that vinblastine, colchicines, and heavy water (D(2)O) did not alter cell deformability, even though vinblastine caused formation of intracellular crystals of microtubular protein, indicates that microtubules are not major determinants of cell deformability. The observations that beads adhered normally to surfaces of cytochalasin B- and of PCMBS-treated cells and that shear-stress induced deformation was increased whereas phagocytosis was markedly inhibited, suggest that deformation of cells around beads associated with ingestion depends on some form of cellular (contractile?) activity, whereas deformation of cells by fluid shear stress is a passive phenomenon.
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- BARANY M., BARANY K. Studies on "active centers" of L-myosin. Biochim Biophys Acta. 1959 Oct;35:293–309. doi: 10.1016/0006-3002(59)90378-6. [DOI] [PubMed] [Google Scholar]
- Bensch K. G., Malawista S. E. Microtubular crystals in mammalian cells. J Cell Biol. 1969 Jan;40(1):95–107. doi: 10.1083/jcb.40.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Böyum A. Isolation of leucocytes from human blood. A two-phase system for removal of red cells with methylcellulose as erythrocyte-aggregating agent. Scand J Clin Lab Invest Suppl. 1968;97:9–29. [PubMed] [Google Scholar]
- Carraway K. L., Shin B. C. Specific modification, isolation, and partial characterization of an erythrocyte membrane protein. J Biol Chem. 1972 Apr 10;247(7):2102–2108. [PubMed] [Google Scholar]
- Cline M. J., Lehrer R. I. Phagocytosis by human monocytes. Blood. 1968 Sep;32(3):423–435. [PubMed] [Google Scholar]
- Fine R. E., Bray D. Actin in growing nerve cells. Nat New Biol. 1971 Nov 24;234(47):115–118. doi: 10.1038/newbio234115a0. [DOI] [PubMed] [Google Scholar]
- KARNOVSKY M. L. Metabolic basis of phagocytic activity. Physiol Rev. 1962 Jan;42:143–168. doi: 10.1152/physrev.1962.42.1.143. [DOI] [PubMed] [Google Scholar]
- KIRSCHNER L. B. FLUORIDE INHIBITION OF SODIUM EXTRUSION FROM SWINE ERYTHROCYTES AND ITS METABOLIS CORRELATES. Arch Biochem Biophys. 1964 Jul 20;106:57–64. doi: 10.1016/0003-9861(64)90156-0. [DOI] [PubMed] [Google Scholar]
- Kane R. E. Preparation and purification of polymerized actin from sea urchin egg extracts. J Cell Biol. 1975 Aug;66(2):305–315. doi: 10.1083/jcb.66.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Knauf P. A., Rothstein A. Chemical modification of membranes. II. Permeation paths for sulfhydryl agents. J Gen Physiol. 1971 Aug;58(2):211–223. doi: 10.1085/jgp.58.2.211. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kvarstein B. The effect of temperature, mabolic inhibitors, andEDTA on pgocytosis of polystyrene latex particles by human leucocytes. Scand J Clin Lab Invest. 1969 Oct;24(3):271–277. doi: 10.3109/00365516909080162. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lichtman M. A. Cellular deformability during maturation of the myeloblast. Possible role in marrow egress. N Engl J Med. 1970 Oct 29;283(18):943–948. doi: 10.1056/NEJM197010292831801. [DOI] [PubMed] [Google Scholar]
- Lichtman M. A., Weed R. I. Alteration of the cell periphery during granulocyte maturation: relationship to cell function. Blood. 1972 Mar;39(3):301–316. [PubMed] [Google Scholar]
- Michell R. H., Pancake S. J., Noseworthy J., Karnovsky M. L. Measurement of rates of phagocytosis: the use of cellular monolayers. J Cell Biol. 1969 Jan;40(1):216–224. doi: 10.1083/jcb.40.1.216. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miller M. E., Myers K. Cellular deformability of the human peripheral blood polymorphonuclear leukocyte: method of study, normal variation and effects of physical and chemical alterations. J Reticuloendothel Soc. 1975 Dec;18(6):337–345. [PubMed] [Google Scholar]
- Perry S. V., Cotterill J. The action of thiol inhibitors on the interaction of F-actin and heavy meromyosin. Biochem J. 1964 Sep;92(3):603–608. doi: 10.1042/bj0920603. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pollard T. D., Ito S. Cytoplasmic filaments of Amoeba proteus. I. The role of filaments in consistency changes and movement. J Cell Biol. 1970 Aug;46(2):267–289. doi: 10.1083/jcb.46.2.267. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spudich J. A., Lin S. Cytochalasin B, its interaction with actin and actomyosin from muscle (cell movement-microfilaments-rabbit striated muscle). Proc Natl Acad Sci U S A. 1972 Feb;69(2):442–446. doi: 10.1073/pnas.69.2.442. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Stossel T. P., Mason R. J., Hartwig J., Vaughan M. Quantitative studies of phagocytosis by polymorphonuclear leukocytes: use of emulsions to measure the initial rate of phagocytosis. J Clin Invest. 1972 Mar;51(3):615–624. doi: 10.1172/JCI106851. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taylor D. L., Condeelis J. S., Moore P. L., Allen R. D. The contractile basis of amoeboid movement. I. The chemical control of motility in isolated cytoplasm. J Cell Biol. 1973 Nov;59(2 Pt 1):378–394. doi: 10.1083/jcb.59.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wilson L., Bryan J., Ruby A., Mazia D. Precipitation of proteins by vinblastine and calcium ions. Proc Natl Acad Sci U S A. 1970 Jul;66(3):807–814. doi: 10.1073/pnas.66.3.807. [DOI] [PMC free article] [PubMed] [Google Scholar]
