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. 1973 Aug 1;58(2):346–356. doi: 10.1083/jcb.58.2.346

QUANTITATIVE STUDIES OF PHAGOCYTOSIS

Kinetic Effects of Cations and Heat-Labile Opsonin

Thomas P Stossel 1
PMCID: PMC2109045  PMID: 4738105

Abstract

Kinetic analysis of the initial ingestion rate of albumin-coated paraffin oil particles by human granulocytes and rabbit alveolar macrophages was undertaken to study the mechanism of action of cations and of heat-labile opsonin on engulfment. The rate of uptake of the particles was stimulated by Ca++, Mg++, Mn++, or Co++. At high concentrations (> 20 mM) Ca++ and Mg++ inhibited the rate of ingestion. Treatment of the particles with fresh serum (heat-labile opsonin) also stimulated the rate of ingestion. 125I-labeled C3 was bound to the particles during opsonization. C3-deficient human serum lacked opsonic activity, which was restored by addition of purified C3. Normal, C2-deficient, and hereditary angioneurotic edema sera had equivalent opsonic activity. The serum opsonic activity thus involved C3 fixation to the particles by means of the properdin system. Although Mg++ and heat-labile opsonin both accelerated the maximal rates of ingestion of the particles, neither altered the particle concentrations associated with one-half maximal ingestion rates. Opsonization of the particles markedly diminished the concentrations of divalent cations causing both stimulatory and inhibitory effects on ingestion rates and altered the shapes of the cation activation curves. 45Ca was not bound to the particles during opsonization. The results are consistent with a mechanism whereby divalent cations and heat-labile opsonin activate ingestion by stimulating the work of engulfment rather than by merely enhancing cell-particle affinity, and whereby heat-labile opsonin acts by potentiating the effects of divalent cations.

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Selected References

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  1. Abramson N., Gelfand E. W., Jandl J. H., Rosen F. S. The interaction between human monocytes and red cells. Specificity for IgG subclasses and IgG fragments. J Exp Med. 1970 Dec 1;132(6):1207–1215. doi: 10.1084/jem.132.6.1207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Alper C. A., Colten H. R., Rosen F. S., Rabson A. R., Macnab G. M., Gear J. S. Homozygous deficiency of C3 in a patient with repeated infections. Lancet. 1972 Dec 2;2(7788):1179–1181. doi: 10.1016/s0140-6736(72)92598-6. [DOI] [PubMed] [Google Scholar]
  3. DONALDSON V. H., ROSEN F. S. ACTION OF COMPLEMENT IN HEREDITARY ANGIONEUROTIC EDEMA: THE ROLE OF C'1-ESTERASE. J Clin Invest. 1964 Nov;43:2204–2213. doi: 10.1172/JCI105094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Gigli I., Nelson R. A., Jr Complement dependent immune phagocytosis. I. Requirements for C'1, C'4, C'2, C'3. Exp Cell Res. 1968 Jul;51(1):45–67. doi: 10.1016/0014-4827(68)90158-4. [DOI] [PubMed] [Google Scholar]
  5. Huber H., Polley M. J., Linscott W. D., Fudenberg H. H., Müller-Eberhard H. J. Human monocytes: distinct receptor sites for the third component of complement and for immunoglobulin G. Science. 1968 Dec 13;162(3859):1281–1283. doi: 10.1126/science.162.3859.1281. [DOI] [PubMed] [Google Scholar]
  6. Johnston R. B., Jr, Klemperer M. R., Alper C. A., Rosen F. S. The enhancement of bacterial phagocytosis by serum. The role of complement components and two cofactors. J Exp Med. 1969 Jun 1;129(6):1275–1290. doi: 10.1084/jem.129.6.1275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Klemperer M. R., Austen K. F., Rosen F. S. Hereditary deficiency of the second component of complement (C'2) in man: further observations on a second kindred. J Immunol. 1967 Jan;98(1):72–78. [PubMed] [Google Scholar]
  8. Korn E. D., Weisman R. A. Phagocytosis of latex beads by Acanthamoeba. II. Electron microscopic study of the initial events. J Cell Biol. 1967 Jul;34(1):219–227. doi: 10.1083/jcb.34.1.219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. MYRVIK Q., LEAKE E. S., FARISS B. Studies on pulmonary alveolar macrophages from the normal rabbit: a technique to procure them in a high state of purity. J Immunol. 1961 Feb;86:128–132. [PubMed] [Google Scholar]
  11. Mason R. J., Stossel T. P., Vaughan M. Quantitative studies of phagocytosis by alveolar macrophages. Biochim Biophys Acta. 1973 May 28;304(3):864–870. doi: 10.1016/0304-4165(73)90233-x. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. NILSSON U. R., MUELLER-EBERHARD H. J. ISOLATION OF BETA IF-GLOBULIN FROM HUMAN SERUM AND ITS CHARACTERIZATION AS THE FIFTH COMPONENT OF COMPLEMENT. J Exp Med. 1965 Aug 1;122:277–298. doi: 10.1084/jem.122.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Nathan D. G., Baehner R. L., Weaver D. K. Failure of nitro blue tetrazolium reduction in the phagocytic vacuoles of leukocytes in chronic granulomatous disease. J Clin Invest. 1969 Oct;48(10):1895–1904. doi: 10.1172/JCI106156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. SBARRA A. J., KARNOVSKY M. L. The biochemical basis of phagocytosis. I. Metabolic changes during the ingestion of particles by polymorphonuclear leukocytes. J Biol Chem. 1959 Jun;234(6):1355–1362. [PubMed] [Google Scholar]
  16. Shibata N., Tatsumi N., Tanaka K., Okamura Y., Senda N. A contractile protein possessing Ca 2+ sensitivity (natural actomyosin) from leucocytes. Its extraction and some of its properties. Biochim Biophys Acta. 1972 Feb 28;256(2):565–576. doi: 10.1016/0005-2728(72)90084-9. [DOI] [PubMed] [Google Scholar]
  17. Shin H. S., Smith M. R., Wood W. B., Jr Heat labile opsonins to pneumococcus. II. Involvement of C3 and C5. J Exp Med. 1969 Dec 1;130(6):1229–1241. doi: 10.1084/jem.130.6.1229. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Smith M. R., Wood W. B., Jr Heat labile opsonins to pneumococcus. I. Participation of complement. J Exp Med. 1969 Dec 1;130(6):1209–1227. doi: 10.1084/jem.130.6.1209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Stossel T. P., Alper C. A., Rosen F. S. Serum-dependent phagocytosis of paraffin oil emulsified with bacterial lipopolysaccharide. J Exp Med. 1973 Mar 1;137(3):690–705. doi: 10.1084/jem.137.3.690. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Stossel T. P., Mason R. J., Pollard T. D., Vaughan M. Isolation and properties of phagocytic vesicles. II. Alveolar macrophages. J Clin Invest. 1972 Mar;51(3):604–614. doi: 10.1172/JCI106850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Weisman R. A., Korn E. D. Phagocytosis of latex beads by Acanthamoeba. I. Biochemical properties. Biochemistry. 1967 Feb;6(2):485–497. doi: 10.1021/bi00854a017. [DOI] [PubMed] [Google Scholar]

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