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. 1977 Apr 1;145(4):931–945. doi: 10.1084/jem.145.4.931

The presence of two Fc receptors on mouse macrophages: evidence from a variant cell line and differential trypsin sensitivity

PMCID: PMC2180633  PMID: 323399

Abstract

A stable variant of a clone of the P388D1 macrophage line was isolated using four cycles of treatment with mouse IgG2a-rabbit anti-kappa complexes and rabbit complement. The variant had the same Ka and about the same number of sites per cell for IgG2a as the parent line. However, the variant had 10% as many binding sites for rabbit IgG in soluble antigen-antibody complexes, and the affinity of binding was threefold higher. This change in binding of complexes to cells of a cloned line without alternation of IgG2a binding provides evidence for the presence of two distinct Fc receptors. The two receptors could also be distiguished on the P388D1 line and on thioglycollate-induced mouse peritoneal macrophages by differential sensitivity to trypsinization. The receptors that bind monomeric IgG2a, sheep erythrocytes (SRBC) covalently bound with IgG2a or rabbit IgG using glutaraldehyde, and Sephadex beads coupled with IgG2a or rabbit IgG using cyanogen bromide activitation, is sensitive to trypsinization. The receptor that binds soluble rabbit antibody-antigen complexes, trinitrophenyl-SRBC and dinitrophenyl(DNP)-bovine serum albumin Sephadex beads coated with rabbit anti-DNP IgG is trypsin resitant, the observation that uncomplexed rabbit IgG oes not bind to the trypsin-resistant receptor, whereas the same IgG bound to its antigen does, suggests that conformational changes induced by the binding of ligand may be of consequence in macrophage function.

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Selected References

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  1. Arend W. P., Mannik M. In vitro adherence of soluble immune complexes to macrophages. J Exp Med. 1972 Sep 1;136(3):514–531. doi: 10.1084/jem.136.3.514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Avrameas S., Taudou B., Chuilon S. Glutaraldehyde, cyanuric chloride and tetrazotized O-dianisidine as coupling reagents in the passive hemagglutination test. Immunochemistry. 1969 Jan;6(1):67–76. doi: 10.1016/0019-2791(69)90179-7. [DOI] [PubMed] [Google Scholar]
  4. Basten A., Miller J. F., Sprent J., Pye J. A receptor for antibody on B lymphocytes. I. Method of detection and functional significance. J Exp Med. 1972 Mar 1;135(3):610–626. doi: 10.1084/jem.135.3.610. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cohen A. M., Burdick J. F., Ketcham A. S. Cell-mediated cytotoxicity: an assay using 125 I-iododeoxyuridine-labeled target cells. J Immunol. 1971 Sep;107(3):895–898. [PubMed] [Google Scholar]
  6. Cuatrecasas P. Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J Biol Chem. 1970 Jun;245(12):3059–3065. [PubMed] [Google Scholar]
  7. Dickler H. B., Kunkel H. G. Interaction of aggregated -globulin with B lymphocytes. J Exp Med. 1972 Jul 1;136(1):191–196. doi: 10.1084/jem.136.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gordon J., Murgita R. A. Suppression and augmentation of the primary in vitro immune response by different classes of antibodies. Cell Immunol. 1975 Feb;15(2):392–402. doi: 10.1016/0008-8749(75)90017-9. [DOI] [PubMed] [Google Scholar]
  9. Griffin F. M., Jr, Bianco C., Silverstein S. C. Characterization of the macrophage receptro for complement and demonstration of its functional independence from the receptor for the Fc portion of immunoglobulin G. J Exp Med. 1975 Jun 1;141(6):1269–1277. doi: 10.1084/jem.141.6.1269. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Griffin F. M., Jr, Griffin J. A., Leider J. E., Silverstein S. C. Studies on the mechanism of phagocytosis. I. Requirements for circumferential attachment of particle-bound ligands to specific receptors on the macrophage plasma membrane. J Exp Med. 1975 Nov 1;142(5):1263–1282. doi: 10.1084/jem.142.5.1263. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Henson P. M. The adherence of leucocytes and platelets induced by fixed IgG antibody or complement. Immunology. 1969 Jan;16(1):107–121. [PMC free article] [PubMed] [Google Scholar]
  12. Howard J. G., Benacerraf B. Properties of macrophage receptors for cytophilic antibodies. Br J Exp Pathol. 1966 Apr;47(2):193–200. [PMC free article] [PubMed] [Google Scholar]
  13. Huber R., Deisenhofer J., Colman P. M., Matsushima M., Palm W. Crystallographic structure studies of an IgG molecule and an Fc fragment. Nature. 1976 Dec 2;264(5585):415–420. doi: 10.1038/264415a0. [DOI] [PubMed] [Google Scholar]
  14. KLEIN G., PERLMANN P. IN VITRO CYTOTOXIC EFFECT OF ISOANTIBODY MEASURED AS ISOTOPE RELEASE FROM LABELLED TARGET CELL DNA. Nature. 1963 Aug 3;199:451–453. doi: 10.1038/199451a0. [DOI] [PubMed] [Google Scholar]
  15. 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]
  16. 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]
  17. 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]
  18. Linscott W. D. Effect of cell surface antigen density on immunological enhancement. Nature. 1970 Nov 28;228(5274):824–827. doi: 10.1038/228824a0. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. NISONOFF A. ENZYMATIC DIGESTION OF RABBIT GAMMA GLOBULIN AND ANTIBODY AND CHROMATOGRAPHY OF DIGESTION PRODUCTS. Methods Med Res. 1964;10:134–141. [PubMed] [Google Scholar]
  21. Rabinovitch M., DeStefano M. J. Cell shape changes induced by cationic anesthetics. J Exp Med. 1976 Feb 1;143(2):290–304. doi: 10.1084/jem.143.2.290. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rabinovitch M. The dissociation of the attachment and ingestion phases of phagocytosis by macrophages. Exp Cell Res. 1967 Apr;46(1):19–28. doi: 10.1016/0014-4827(67)90405-3. [DOI] [PubMed] [Google Scholar]
  23. Rhodes J. Macrophage heterogeneity in receptor activity: the activation of macrophage Fc receptor function in vivo and in vitro. J Immunol. 1975 Mar;114(3):976–981. [PubMed] [Google Scholar]
  24. Rittenberg M. B., Pratt K. L. Antitrinitrophenyl (TNP) plaque assay. Primary response of Balb/c mice to soluble and particulate immunogen. Proc Soc Exp Biol Med. 1969 Nov;132(2):575–581. doi: 10.3181/00379727-132-34264. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. 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]
  28. Tigelaar R. E., Vaz N. M., Ovary Z. Immunoglobulin receptors on mouse mast cells. J Immunol. 1971 Mar;106(3):661–672. [PubMed] [Google Scholar]
  29. Unkeless J. C., Eisen H. N. Binding of monomeric immunoglobulins to Fc receptors of mouse macrophages. J Exp Med. 1975 Dec 1;142(6):1520–1533. doi: 10.1084/jem.142.6.1520. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. 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]
  31. WATKINS J. F. ADSORPTION OF SENSITIZED SHEEP ERYTHROCYTES TO HELA CELLS INFECTED WITH HERPES SIMPLEX VIRUS. Nature. 1964 Jun 27;202:1364–1365. doi: 10.1038/2021364a0. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Walker W. S. Functional heterogeneity of macrophages: subclasses of peritoneal macrophages with different antigen-binding activities and immune complex receptors. Immunology. 1974 May;26(5):1025–1037. [PMC free article] [PubMed] [Google Scholar]
  34. Walker W. S. Separate Fc-receptors for immunoglogulins IgG2a and IgG2b on an established cell line of mouse macrophages. J Immunol. 1976 Apr;116(4):911–914. [PubMed] [Google Scholar]

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