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J Exp Med. 1989 August 1; 170(2): 511–526.
PMCID: PMC2189400

Antibody-dependent antitumor cytotoxicity by human monocytes cultured with recombinant macrophage colony-stimulating factor. Induction of efficient antibody-mediated antitumor cytotoxicity not detected by isotope release assays


Macrophage colony-stimulating factor (M-CSF) is known to stimulate proliferation of monocyte/macrophage progenitors and enhance in vitro antitumor cytotoxicity by murine macrophages. In this paper we have shown that recombinant human M-CSF causes human peripheral blood monocytes to differentiate in culture into metabolically active macrophage-like cells. These cells mediate very efficient antibody- dependent cellular cytotoxicity (ADCC) against human melanoma and neuroblastoma cell lines in the presence of two murine IgG3 mAbs (3F8 and R24). They also mediate antibody-independent cytotoxicity (or cytostasis) to a lesser extent. Human serum had an inconsistent effect on ADCC, but often induced similar high levels of ADCC. Cytotoxicity was measured using a novel ELISA to detect surviving tumor cells after ADCC. Two conventional isotope-release assays (51Cr and [3H]TdR) underestimated or entirely failed to detect ADCC by M-CSF-activated monocytes. Optimal activation occurred with 100-300 U/ml of M-CSF, and required 9-11 d for completion. Most of the M-CSF cultured monocytes expressed the low-affinity Fc receptor (CD16). ADCC by cells of the monocyte/macrophage lineage using murine IgG3 mAbs may have significance for the immunotherapy of human malignancies.

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Selected References

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  • Wong GG, Temple PA, Leary AC, Witek-Giannotti JS, Yang YC, Ciarletta AB, Chung M, Murtha P, Kriz R, Kaufman RJ, et al. Human CSF-1: molecular cloning and expression of 4-kb cDNA encoding the human urinary protein. Science. 1987 Mar 20;235(4795):1504–1508. [PubMed]
  • Tushinski RJ, Oliver IT, Guilbert LJ, Tynan PW, Warner JR, Stanley ER. Survival of mononuclear phagocytes depends on a lineage-specific growth factor that the differentiated cells selectively destroy. Cell. 1982 Jan;28(1):71–81. [PubMed]
  • Chen BD, Mueller M, Chou TH. Role of granulocyte/macrophage colony-stimulating factor in the regulation of murine alveolar macrophage proliferation and differentiation. J Immunol. 1988 Jul 1;141(1):139–144. [PubMed]
  • Ralph P, Warren MK, Nakoinz I, Lee MT, Brindley L, Sampson-Johannes A, Kawasaki ES, Ladner MB, Strickler JE, Boosman A, et al. Biological properties and molecular biology of the human macrophage growth factor, CSF-1. Immunobiology. 1986 Sep;172(3-5):194–204. [PubMed]
  • Ralph P, Nakoinz I. Stimulation of macrophage tumoricidal activity by the growth and differentiation factor CSF-1. Cell Immunol. 1987 Apr 1;105(2):270–279. [PubMed]
  • Nakoinz I, Ralph P. Stimulation of macrophage antibody-dependent killing of tumor targets by recombinant lymphokine factors and M-CSF. Cell Immunol. 1988 Oct 15;116(2):331–340. [PubMed]
  • Ampel NM, Wing EJ, Waheed A, Shadduck RK. Stimulatory effects of purified macrophage colony-stimulating factor on murine resident peritoneal macrophages. Cell Immunol. 1986 Feb;97(2):344–356. [PubMed]
  • Wing EJ, Ampel NM, Waheed A, Shadduck RK. Macrophage colony-stimulating factor (M-CSF) enhances the capacity of murine macrophages to secrete oxygen reduction products. J Immunol. 1985 Sep;135(3):2052–2056. [PubMed]
  • Warren MK, Ralph P. Macrophage growth factor CSF-1 stimulates human monocyte production of interferon, tumor necrosis factor, and colony stimulating activity. J Immunol. 1986 Oct 1;137(7):2281–2285. [PubMed]
  • Fidler IJ. Macrophages and metastasis--a biological approach to cancer therapy. Cancer Res. 1985 Oct;45(10):4714–4726. [PubMed]
  • Grabstein KH, Urdal DL, Tushinski RJ, Mochizuki DY, Price VL, Cantrell MA, Gillis S, Conlon PJ. Induction of macrophage tumoricidal activity by granulocyte-macrophage colony-stimulating factor. Science. 1986 Apr 25;232(4749):506–508. [PubMed]
  • Sampson-Johannes A, Carlino JA. Enhancement of human monocyte tumoricidal activity by recombinant M-CSF. J Immunol. 1988 Nov 15;141(10):3680–3686. [PubMed]
  • Johnson WD, Jr, Mei B, Cohn ZA. The separation, long-term cultivation, and maturation of the human monocyte. J Exp Med. 1977 Dec 1;146(6):1613–1626. [PMC free article] [PubMed]
  • Nakagawara A, Nathan CF, Cohn ZA. Hydrogen peroxide metabolism in human monocytes during differentiation in vitro. J Clin Invest. 1981 Nov;68(5):1243–1252. [PMC free article] [PubMed]
  • Musson RA. Human serum induces maturation of human monocytes in vitro. Changes in cytolytic activity, intracellular lysosomal enzymes, and nonspecific esterase activity. Am J Pathol. 1983 Jun;111(3):331–340. [PubMed]
  • Becker S, Warren MK, Haskill S. Colony-stimulating factor-induced monocyte survival and differentiation into macrophages in serum-free cultures. J Immunol. 1987 Dec 1;139(11):3703–3709. [PubMed]
  • Eliason JF. Granulocyte-macrophage colony formation in serum-free culture: effects of purified colony-stimulating factors and modulation by hydrocortisone. J Cell Physiol. 1986 Aug;128(2):231–238. [PubMed]
  • Adams DO, Hall T, Steplewski Z, Koprowski H. Tumors undergoing rejection induced by monoclonal antibodies of the IgG2a isotype contain increased numbers of macrophages activated for a distinctive form of antibody-dependent cytolysis. Proc Natl Acad Sci U S A. 1984 Jun;81(11):3506–3510. [PubMed]
  • Masui H, Moroyama T, Mendelsohn J. Mechanism of antitumor activity in mice for anti-epidermal growth factor receptor monoclonal antibodies with different isotypes. Cancer Res. 1986 Nov;46(11):5592–5598. [PubMed]
  • Steplewski Z, Lubeck MD, Koprowski H. Human macrophages armed with murine immunoglobulin G2a antibodies to tumors destroy human cancer cells. Science. 1983 Aug 26;221(4613):865–867. [PubMed]
  • Johnson WJ, Steplewski Z, Matthews TJ, Hamilton TA, Koprowski H, Adams DO. Cytolytic interactions between murine macrophages, tumor cells, and monoclonal antibodies: characterization of lytic conditions and requirements for effector activation. J Immunol. 1986 Jun 15;136(12):4704–4713. [PubMed]
  • Herlyn D, Herlyn M, Steplewski Z, Koprowski H. Monoclonal anti-human tumor antibodies of six isotypes in cytotoxic reactions with human and murine effector cells. Cell Immunol. 1985 Apr 15;92(1):105–114. [PubMed]
  • Thurin J, Thurin M, Kimoto Y, Herlyn M, Lubeck MD, Elder DE, Smereczynska M, Karlsson KA, Clark WM, Jr, Steplewski Z, et al. Monoclonal antibody-defined correlations in melanoma between levels of GD2 and GD3 antigens and antibody-mediated cytotoxicity. Cancer Res. 1987 Mar 1;47(5):1229–1233. [PubMed]
  • Gardner CR, Wasserman AJ, Laskin DL. Differential sensitivity of tumor targets to liver macrophage-mediated cytotoxicity. Cancer Res. 1987 Dec 15;47(24 Pt 1):6686–6691. [PubMed]
  • Calafat J, Janssen H, Hekman A. Mouse monoclonal antibodies direct phagocytosis of tumor cells by human monocytes. Leuk Res. 1986;10(11):1347–1351. [PubMed]
  • Cheung NK, Saarinen UM, Neely JE, Landmeier B, Donovan D, Coccia PF. Monoclonal antibodies to a glycolipid antigen on human neuroblastoma cells. Cancer Res. 1985 Jun;45(6):2642–2649. [PubMed]
  • Cheung NK, Walter EI, Smith-Mensah WH, Ratnoff WD, Tykocinski ML, Medof ME. Decay-accelerating factor protects human tumor cells from complement-mediated cytotoxicity in vitro. J Clin Invest. 1988 Apr;81(4):1122–1128. [PMC free article] [PubMed]
  • Gmelig-Meyling F, Waldmann TA. Separation of human blood monocytes and lymphocytes on a continuous Percoll gradient. J Immunol Methods. 1980;33(1):1–9. [PubMed]
  • Cheresh DA, Pierschbacher MD, Herzig MA, Mujoo K. Disialogangliosides GD2 and GD3 are involved in the attachment of human melanoma and neuroblastoma cells to extracellular matrix proteins. J Cell Biol. 1986 Mar;102(3):688–696. [PMC free article] [PubMed]
  • Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983 Dec 16;65(1-2):55–63. [PubMed]
  • Munn DH, Cheung NK. Interleukin-2 enhancement of monoclonal antibody-mediated cellular cytotoxicity against human melanoma. Cancer Res. 1987 Dec 15;47(24 Pt 1):6600–6605. [PubMed]
  • Kilbourn RG, Klostergaard J, Lopez-Berestein G. Activated macrophages secrete a soluble factor that inhibits mitochondrial respiration of tumor cells. J Immunol. 1984 Nov;133(5):2577–2581. [PubMed]
  • Johnson WJ, Bolognesi DP, Adams DO. Antibody-dependent cytolysis (ADCC) of tumor cells by activated murine macrophages is a two-step process: quantification of target binding and subsequent target lysis. Cell Immunol. 1984 Jan;83(1):170–180. [PubMed]
  • Klostergaard J. Role of tumor necrosis factor in monocyte/macrophage tumor cytotoxicity in vitro. Nat Immun Cell Growth Regul. 1987;6(4):161–166. [PubMed]
  • Fleit HB, Wright SD, Unkeless JC. Human neutrophil Fc gamma receptor distribution and structure. Proc Natl Acad Sci U S A. 1982 May;79(10):3275–3279. [PubMed]
  • Clarkson SB, Ory PA. CD16. Developmentally regulated IgG Fc receptors on cultured human monocytes. J Exp Med. 1988 Feb 1;167(2):408–420. [PMC free article] [PubMed]
  • Perussia B, Trinchieri G, Jackson A, Warner NL, Faust J, Rumpold H, Kraft D, Lanier LL. The Fc receptor for IgG on human natural killer cells: phenotypic, functional, and comparative studies with monoclonal antibodies. J Immunol. 1984 Jul;133(1):180–189. [PubMed]
  • Simmons D, Seed B. The Fc gamma receptor of natural killer cells is a phospholipid-linked membrane protein. Nature. 1988 Jun 9;333(6173):568–570. [PubMed]
  • Bartocci A, Mastrogiannis DS, Migliorati G, Stockert RJ, Wolkoff AW, Stanley ER. Macrophages specifically regulate the concentration of their own growth factor in the circulation. Proc Natl Acad Sci U S A. 1987 Sep;84(17):6179–6183. [PubMed]
  • Saarinen UM, Coccia PF, Gerson SL, Pelley R, Cheung NK. Eradication of neuroblastoma cells in vitro by monoclonal antibody and human complement: method for purging autologous bone marrow. Cancer Res. 1985 Nov;45(11 Pt 2):5969–5975. [PubMed]
  • Cheung NK, Lazarus H, Miraldi FD, Abramowsky CR, Kallick S, Saarinen UM, Spitzer T, Strandjord SE, Coccia PF, Berger NA. Ganglioside GD2 specific monoclonal antibody 3F8: a phase I study in patients with neuroblastoma and malignant melanoma. J Clin Oncol. 1987 Sep;5(9):1430–1440. [PubMed]
  • Gauci CL, Alexander P. The macrophage content of some human tumours. Cancer Lett. 1975 Sep;1(1):29–32. [PubMed]
  • McBride WH. Phenotype and functions of intratumoral macrophages. Biochim Biophys Acta. 1986 Aug 5;865(1):27–41. [PubMed]

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