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. 1994 May;39(3):179–184. doi: 10.1007/BF01533384

Enhanced killing capacity of human Kupffer cells after activation with human granulocyte/macrophage-colony-stimulating factor and interferon γ

B Schuurman 1, G Heuff 1, R H J Beelen 2, S Meyer 1,✉
PMCID: PMC11038622  PMID: 7923248

Abstract

In this study we investigated the effect of the cytokines human granulocyte/macrophage-colony-stimulating Factor (hGM-CSF) and interferon γ (IFNγ) on human Kupffer-cell-mediated cytotoxicity against the SW948 coloncarcinoma cell line. Kupffer cells were isolated from small liver wedge biopsies, taken from 14 patient who had had abdominal surgery for colon carcinoma or partial hepatectomy. The cells were incubated with hGM-CSF (100 ng/ml), or with IFNγ (100 U/ml) or with their combination and the perecentage cytotoxicity was determined using a recently described modified assay. Additional experiments were performed with tumour-necrosis-factor-α(TNFα)-sensitive U937 cells as target. The TNFα secretion of Kupffer cells was measured and we evaluated the effect of TNFα on colon tumour targets. We performed human-Kupffer cell-mediated cytotoxicity blocking experiments with anti-TNFα and used paraformaldehydefixed Kupffer cells to demonstrate lysis of TNFα-sensitive WEHI-164 cells and of SW948 cells. The overall cytotoxicity against SW948 caused by unactivated Kupffer cells (n=14), and by Kupffer cells activated with hGM-CSF (n=14), IFNγ (n=6) or their combination (n=6) was respectively: 19.5±2.6%, 25.3±2.9% 41±9.4% and 45.6±8% at E/T=1 and 28.2±2.9%, 35.6±3.2%, 55.6±9.7% and 62.8% at E/T=5. All differences were statistically significant (P<0.05). No growth-promoting activity by hGM-CSF on the SW948 tumour cells was observed. U937 cells were highly susceptible to Kupffer-cell-mediated cytotoxicity. The TNFα secretion by human Kupffer cells increased in parallel to their cytotoxicity after incubation with these cytokines. Soluble TNFα had only a slight anti-proliferative effect on SW948 cells, while specific anti-TNFα blocked Kupffer cell cytotoxicity by up to 80%. Finally, paraformaldehyde-fixed Kupffer cells were able to lyse WEHI-164 and SW948 cells. This indicates that expression of cell-associated TNFα is the main cytolytic mechanism of human-Kupffer-cell-mediated cytotoxicity. The implications for the use of hGM-CSF and IFNγ in vivo are discussed.

Key words: Kupffer cell, Human granulocyte/macrophage-colony-stimulating factor, Interferon γ, Cytotoxicity, SW948, Tumor necrosis factor α

References

  • 1.Adams DO, Nathan CF. Molecular mechanisms in tumor-cell killing by activated macrophages. Immunol Today. 1989;10:92. doi: 10.1016/0167-5699(83)90005-1. [DOI] [PubMed] [Google Scholar]
  • 2.Baldwin GC, Gasson JC, Kaufman SE, Quan SG, Williams RE, Avalos BR, Gazdar AF, Golde DW, DiPersio JF. Non-hematopoietic tumor cells express functional GM-CSF receptors. Blood. 1989;73:1033. [PubMed] [Google Scholar]
  • 3.Curley SA, Roh MS, Kleinerman E, Klostergaard J. Human recombinant macrophage colony stimulating factor activates murine Kupffer cells to a cytotoxic state. Lymphokine Res. 1990;9:355. [PubMed] [Google Scholar]
  • 4.Daemen T. Activation of Kuffer cell tumoricidal activity by immunomodulators encapsulated in liposomes. Res Immunol. 1992;143:211. doi: 10.1016/s0923-2494(92)80168-k. [DOI] [PubMed] [Google Scholar]
  • 5.Daemen T, Veninga A, Roerdink FH, Scherphof GL. In vitro activation of rat livermacrophages to tumoricidal state by free or liposome encapsulated muramyl di-peptide. Cancer Res. 1986;46:4330. [PubMed] [Google Scholar]
  • 6.Decker T, Lohmann-Matthes ML, Gifford GE. Cell associated tumor necrosis factor (TNF) as a killing mechanism of activated cytotoxic macrophages. J Immunol. 1987;138:957. [PubMed] [Google Scholar]
  • 7.Essner R, Rhoades K, McBride WH, Morton DL, Economou JS. Differential effect of granulocyte-macrophage colony stimulating factor and macrophage colony stimulating factor on tumor necrosis factor and interleukin-1 production in human monocytes. J Clin Lab Immunol. 1990;32:161. [PubMed] [Google Scholar]
  • 8.Grabstein KH, Urdal KH, Tushinsky RJ, Mochizuki DY, Price VL, Cantrell MA, Gillis S, Conlon PJ. Induction of macrophage tumoricidal activity by granulocyte-macrophage colony stimulating factor. Science. 1986;232:506. doi: 10.1126/science.3083507. [DOI] [PubMed] [Google Scholar]
  • 9.Hamilton JA. Review: colony stimulating factors, cytokines and monocyte-macrophages — some controversies. Immunol Today. 1993;14:18. doi: 10.1016/0167-5699(93)90319-G. [DOI] [PubMed] [Google Scholar]
  • 10.Heuff G, Steenbergen JJE, Van de Loosdrecht AA, Sirovich I, Dijkstra CD, Meyer S, Beelen RHJ. Isolation of cytotoxic Kupffer cells by a modified enzymatic assay: a methodological study. J Immunol Methods. 1993;159:115. doi: 10.1016/0022-1759(93)90148-z. [DOI] [PubMed] [Google Scholar]
  • 11.Heuff G, Oldenburg HSA, Boutkan H, Beelen RHJ, Van Rooijen N, Dijkstra CD, Meyer S. Enhanced tumour growth after selective elimination of Kupffer cells. Cancer Immunol Immunother. 1993;37:125. doi: 10.1007/BF01517045. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.Heuff G, van der Ende MB, Boutkan H, Prevoo W, Visser JJ, Fleuren GJ, Beelen RHJ, Meyer S, Dijkstra CD. Macrophage populations in different stages of induced metastases in rats: an immunohistochemical analysis. Scand J Immunol. 1993;38:10. doi: 10.1111/j.1365-3083.1993.tb01688.x. [DOI] [PubMed] [Google Scholar]
  • 13.Heuff G, Van de Loosdrecht AA, Betjes M, Beelen RHJ, Meyer S (1993) Isolation and purification of large quantities of fresh human Kupffer cells which are cytotoxic against coloncarcinoma (in press) [PubMed]
  • 14.Hill AKD, Redmond HP, Austin OM, Grace PA, Bouchier-Hayes D. Gramulocyte-macrophage colony stimulating factor inhibits tumour growth. Br J Surg. 1993;80:1543. doi: 10.1002/bjs.1800801216. [DOI] [PubMed] [Google Scholar]
  • 15.Inamura N, Sone S, Okubo A, Singh SM, Ogura T. Heterogeneity of human blood monocytes to granulocyte-macrophage colony stimulating factor. J Leukoc Biol. 1990;47:528. doi: 10.1002/jlb.47.6.528. [DOI] [PubMed] [Google Scholar]
  • 16.Kelly PMA, Bliss E, Morton JA, Burns J, McGee JO'D. Monoclonal antibody EBM/11: high cellular specificity for human macrophages. J Clin Pathol. 1988;41:510. doi: 10.1136/jcp.41.5.510. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Kleinerman ES, Knowles RD, Lachman LB, Gutterman JU. Effect of recombinant granulocyte-macrophage colony stimulating factor on human monocyte activity in vitro and following intravenous administration. Cancer Res. 1988;48:2604. [PubMed] [Google Scholar]
  • 18.Leibovitz A, Stinson JC, McCombs WB, McCoy CE, Mazur KC, Mabry ND. Classification of human colorectal adenocarcinoma cell lines. Cancer Res. 1976;36:4562. [PubMed] [Google Scholar]
  • 19.Lieschke GJ, Maher D, Cebon J, et al. Effects of bacterially synthesized recombinant human granulocyte-macrophage colony stimulating factor. Ann Int Med. 1989;110:357. doi: 10.7326/0003-4819-110-5-357. [DOI] [PubMed] [Google Scholar]
  • 20.Metcalf D, Begley CG, Williamson DJ, et al. Hemopoietic responses in mice injected with purified recombinant murine GMCSF. Exp Hematol. 1987;15:1. [PubMed] [Google Scholar]
  • 21.Niskanen E. Hematopoietic growth factors in clinical hematology. Ann Med. 1991;23:615. doi: 10.3109/07853899109148093. [DOI] [PubMed] [Google Scholar]
  • 22.Philips NC. Kupffer cells and liver metastasis. Cancer Metastasis Rev. 1989;8:231. doi: 10.1007/BF00047339. [DOI] [PubMed] [Google Scholar]
  • 23.Ragnhammar P, Masucci G, Frödin JE, Hjelm AL, Mellstedt H. Cytotoxic functions of blood mononuclear cells in patients with colorectal carcinoma treated with mAb 17-1A and granulocyte macrophage-colony-stimulating factor. Cancer Immunol Immunocher. 1992;35:158. doi: 10.1007/BF01756182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Roh MS, Wang L, Oyedeji C, Leroux ME, Curley SA, Pllock RE, Klostergaard J. Human Kupffer cells are cytotoxic against human colon adenocarcinoma. Surgery. 1990;108:400. doi: 10.1002/bjs.1800770937. [DOI] [PubMed] [Google Scholar]
  • 25.Sone S, Lopez-Berestein G, Fidler IJ. Kinetics and function of cytotoxic factors produced by human blood monocytes activated to the tumoricidal state. J Natl Cancer Inst. 1985;74:583. [PubMed] [Google Scholar]
  • 26.Suk K, Somers SD, Erickson KL. Regulation of murine macrophage function by IL-4: IL-4 and IFN-γ differentially regulate macrophage tumoricidal activation. Immunology. 1993;80:617. [PMC free article] [PubMed] [Google Scholar]
  • 27.Twentyman PR, Wright KA. Failure of GM-CSF to influence growth of small cell and non small cell lung cancer cell lines in vitro. Eur J Cancer. 1991;27:6. doi: 10.1016/0277-5379(91)90049-j. [DOI] [PubMed] [Google Scholar]
  • 28.Van de Loosdrecht AA, Nennie E, Ossenkoppcle GJ, Beelen RHJ, Langenhuijsen MMAC. Cell mediated cytotoxicity against U-937 cells by human monocytes and macrophages in a modified colorimetric MTT assay: a methodological study. J Immunol Methods. 1991;141:15. doi: 10.1016/0022-1759(91)90205-t. [DOI] [PubMed] [Google Scholar]
  • 29.Van de Loosdrecht AAL, Ossenkoppele GJ, Beelen RHJ, Broekhoven MG, Langenhuijsen MMAC. Role of interferon-τ and tumor necrosis factor-α in monocyte mediated cytostasis and cytotoxicity against a human histiocytic lymphoma cell line. Cancer Immunol Immunother. 1992;34:393. doi: 10.1007/BF01741750. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Van de Loosdrecht AA, Beelen RHJ, Ossenkoppele GJ, Broekhoven MG, Langenhuijsen MMAC. Cellular and cytokine dependent monocyte mediated leukemic cell death by interferon-γ and tumor necrosis factor-α. Exp Hematol. 1993;21:461. [PubMed] [Google Scholar]
  • 31.Van de Loosdrecht AAL, Ossenkoppele GJ, Beelen RHJ, Broekhoven MG, Langenhuijsen MMAC (1993) Cell cycle specific effects of tumor necrosis factor-α in monocyte mediated leukemic cell death and the role of B16β-integrins. Cancer Res (in press) [PubMed]
  • 32.Van de Winkel JGJ, Anderson CL. Biology of human immunoglobulin G Fe receptors. J Leukoc Biol. 1991;49:511. doi: 10.1002/jlb.49.5.511. [DOI] [PubMed] [Google Scholar]
  • 33.Wing JW, Magee DM, Whiteside TL, Kaplan SS, Shadduk RK. Recombinant human granulocyte-macrophage colony stimulating factor enhances monocyte cytotoxicity and secretion of tumor necrosis factor and interferon in cancer patients. Blood. 1989;73:643. [PubMed] [Google Scholar]
  • 34.Young DA, Lowe LD, Clark SC. Comparison of the effects of IL-3, granulocyte-macrophage colony stimulating factor, and macrophage colony stimulating factor in supporting monocyte differentiation in culture. J Immunol. 1990;145:607. [PubMed] [Google Scholar]

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