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. 1975 Dec 1;142(6):1520–1533. doi: 10.1084/jem.142.6.1520

Binding of monomeric immunoglobulins to Fc receptors of mouse macrophages

PMCID: PMC2190081  PMID: 1194857

Abstract

The binding properties of surface receptors of immunoglobulins on mouse macrophages were studied with mouse myeloma proteins and normal peritoneal macrophages, thioglycollate-stimulated macrophages, and a macrophage cell line, P388D1. Primary cultures of mouse embryo fibroblasts served as controls. IgG2a proteins were bound strongly;IgG2b was bound weakly (one-twentieth as well as IgG2a);IgM, IgA, and IgG1 were not bound significantly. The number of binding sites per cell for IgG2a was 4 X 10(5) for thioglycollate-stimulated cells and 1 X 10(5) for normal and P388D1 cells. Binding was exothermal: with decreasing temperature the equilibrium (association) constants increased and dissociation rate constants decreased (at 37degreesC the respective values were 2 X 10(7) M-1 and 0.26 min-1, the latter value corresponds to a half time for dissociation of 2.6 min). From the rapidity of association and dissociation, it appears that the surface of the macrophage is in a dynamic equilibrium with IgG2a molecules in the cell's immediate microenvironment. The receptors for IgG2a are clearly specific for determinants in the immunoglobulin constant domain: two IgG2a proteins with greatly different isoelectric points (determined by isoelectric focusing) were bound with the same affinity to the same receptors; moreover, the Fc fragment was bound and Fab fragments were not. The Fc receptors for IgG2a proteins were readily eliminated by exposing macrophages briefly to trypsin. The receptors were regenerated during subsequent cultivation in serum-free medium; regeneration was inhibited totally by cycloheximide and partially by actinomycin D.

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Selected References

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  1. Askenase P. W., Hayden B. J. Cytophilic antibodies in mice contact-sensitized with oxazolone. Immunochemical characterization and preferential binding to a trypsin-sensitive macrophage receptor. Immunology. 1974 Oct;27(4):563–576. [PMC free article] [PubMed] [Google Scholar]
  2. Berken A., Benacerraf B. Properties of antibodies cytophilic for macrophages. J Exp Med. 1966 Jan 1;123(1):119–144. doi: 10.1084/jem.123.1.119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Dandliker W. B., Levison S. A. Investigation of antigen-antibody kinetics by fluorescence polarization. Immunochemistry. 1968 Mar;5(2):171–183. doi: 10.1016/0019-2791(68)90101-8. [DOI] [PubMed] [Google Scholar]
  4. Davey M. J., Asherson G. L. Cytophilic antibody. I. Nature of the macrophage receptor. Immunology. 1967 Jan;12(1):13–20. [PMC free article] [PubMed] [Google Scholar]
  5. Evans R., Alexander P. Role of macrophages in tumour immunity. I. Co-operation between macrophages and lymphoid cells in syngeneic tumour immunity. Immunology. 1972 Oct;23(4):615–626. [PMC free article] [PubMed] [Google Scholar]
  6. Evans R., Alexander P. Role of macrophages in tumour immunity. II. Involvement of a macrophage cytophilic factor during syngeneic tumour growth inhibition. Immunology. 1972 Oct;23(4):627–636. [PMC free article] [PubMed] [Google Scholar]
  7. Hiramoto R., Ghanta V. K., McGhee J. R., Schrohenloher R., Hamlin N. M. Use of dextran conjugated columns for the isolation of large quantities of MOPC 104E IgM. Immunochemistry. 1972 Dec;9(12):1251–1253. doi: 10.1016/0019-2791(72)90300-x. [DOI] [PubMed] [Google Scholar]
  8. Inchley C., Grey H. M., Uhr J. W. The cytophilic activity of human immunoglobulins. J Immunol. 1970 Aug;105(2):362–369. [PubMed] [Google Scholar]
  9. Koren H. S., Handwerger B. S., Wunderlich J. R. Identification of macrophage-like characteristics in a cultured murine tumor line. J Immunol. 1975 Feb;114(2 Pt 2):894–897. [PubMed] [Google Scholar]
  10. Kossard S., Nelson D. S. Studies on cytophilic antibodies. IV. The effects of proteolytic enzymes (trypsin and papain) on the attachment to macrophages of cytophilic antibodies. Aust J Exp Biol Med Sci. 1968 Feb;46(1):63–71. [PubMed] [Google Scholar]
  11. LAKI K. The polymerization of proteins; the action of thrombin on fibrinogen. Arch Biochem Biophys. 1951 Jul;32(2):317–324. doi: 10.1016/0003-9861(51)90277-9. [DOI] [PubMed] [Google Scholar]
  12. Lay W. H., Nussenzweig V. Ca++-dependent binding of antigen-19 S antibody complexes to macrophages. J Immunol. 1969 May;102(5):1172–1178. [PubMed] [Google Scholar]
  13. Leslie R. G., Cohen S. Cytophilic activity of IgG2 from sera of guinea-pigs immunized with bovine gamma-globulin. Immunology. 1974 Oct;27(4):589–599. [PMC free article] [PubMed] [Google Scholar]
  14. Leslie R. G., Cohen S. Cytophilic activity of IgG2 from sera of unimmunized guinea-pigs. Immunology. 1974 Oct;27(4):577–587. [PMC free article] [PubMed] [Google Scholar]
  15. Linden C. D., Wright K. L., McConnell H. M., Fox C. F. Lateral phase separations in membrane lipids and the mechanism of sugar transport in Escherichia coli. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2271–2275. doi: 10.1073/pnas.70.8.2271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. LoBuglio A. F., Cotran R. S., Jandl J. H. Red cells coated with immunoglobulin G: binding and sphering by mononuclear cells in man. Science. 1967 Dec 22;158(3808):1582–1585. doi: 10.1126/science.158.3808.1582. [DOI] [PubMed] [Google Scholar]
  17. PORTER R. R. The hydrolysis of rabbit y-globulin and antibodies with crystalline papain. Biochem J. 1959 Sep;73:119–126. doi: 10.1042/bj0730119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Schlessinger J., Steinberg I. Z., Givol D., Hochman J., Pecht I. Antigen-induced conformational changes in antibodies and their Fab fragments studied by circular polarization of fluorescence. Proc Natl Acad Sci U S A. 1975 Jul;72(7):2775–2779. doi: 10.1073/pnas.72.7.2775. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Sonoda S., Schlamowitz M. Studies of 125I trace labeling of immunoglobulin G by chloramine-T. Immunochemistry. 1970 Nov;7(11):885–898. doi: 10.1016/0019-2791(70)90051-0. [DOI] [PubMed] [Google Scholar]
  20. Stanners C. P., Eliceiri G. L., Green H. Two types of ribosome in mouse-hamster hybrid cells. Nat New Biol. 1971 Mar 10;230(10):52–54. doi: 10.1038/newbio230052a0. [DOI] [PubMed] [Google Scholar]
  21. Steinman R. M., Cohn Z. A. The interaction of particulate horseradish peroxidase (HRP)-anti HRP immune complexes with mouse peritoneal macrophages in vitro. J Cell Biol. 1972 Dec;55(3):616–634. doi: 10.1083/jcb.55.3.616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Unkeless J. C., Gordon S., Reich E. Secretion of plasminogen activator by stimulated macrophages. J Exp Med. 1974 Apr 1;139(4):834–850. doi: 10.1084/jem.139.4.834. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Walker W. S., Demus A. Antibody-dependent cytolysis of chicken erythrocytes by an in vitro-established line of mouse peritoneal macrophages. J Immunol. 1975 Feb;114(2 Pt 2):765–769. [PubMed] [Google Scholar]

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