Cancer Immunology, Immunotherapy

, Volume 62, Issue 6, pp 1073–1082 | Cite as

Effects of interleukin-18 on natural killer cells: costimulation of activation through Fc receptors for immunoglobulin

  • Shivani Srivastava
  • David Pelloso
  • Hailin Feng
  • Larry Voiles
  • David Lewis
  • Zdenka Haskova
  • Margaret Whitacre
  • Stephen Trulli
  • Yi-Jiun Chen
  • John Toso
  • Zdenka L. Jonak
  • Hua-Chen Chang
  • Michael J. Robertson
Original Article


The antitumor activity of monoclonal antibodies is mediated by effector cells, such as natural killer (NK) cells, that express Fc receptors for immunoglobulin. Efficacy of monoclonal antibodies, including the CD20 antibody rituximab, could be improved by agents that augment the function of NK cells. Interleukin (IL)-18 is an immunostimulatory cytokine that has antitumor activity in preclinical models. The effects of IL-18 on NK cell function mediated through Fcγ receptors were examined. Human NK cells stimulated with immobilized IgG in vitro secreted IFN-γ as expected; such IFN-γ production was partially inhibited by blocking CD16 with monoclonal antibodies. IL-18 augmented IFN-γ production by NK cells stimulated with immobilized IgG or CD16 antibodies. NK cell IFN-γ production in response to immobilized IgG and/or IL-18 was inhibited by chemical inhibitors of Syk and several other kinases involved in CD16 signaling pathways. IL-18 augmented antibody-dependent cellular cytotoxicity (ADCC) of human NK cells against rituximab-coated Raji cells in vitro. IL-18 and rituximab acted synergistically to promote regression of human lymphoma xenografts in SCID mice. Inasmuch as IL-18 costimulates IFN-γ production and ADCC of NK cells activated through Fc receptors in vitro and augments antitumor activity of rituximab in vivo, it is an attractive cytokine to combine with monoclonal antibodies for treatment of human cancer.


Cancer immunotherapy Cytokines Monoclonal antibodies Lymphoma Rituximab 



This work was supported in part by National Institutes of Health Grant RO1 CA118118 (Michael J. Robertson), Walther Scholar Grant (Shivani Srivastava) from the Indiana University Simon Cancer Center (P30 CA82709) and American Cancer Society IRG (Hua-Chen Chang and Shivani Srivastava). The authors thank Menggang Yu, Ph.D. and Sandra Althouse for statistical support and Lisa Wood, R. N. and nurses in the Indiana CTSI Clinical Research Center for assistance in collection of patient samples.

Conflict of interest

Zdenka Haskova, Margaret Whitacre, Stephen Trulli, Yi-Jiun Chen, John Toso, Zdenka L. Jonak are employees and shareholders of GlaxoSmithKline. All other authors declare that they have no conflict of interest.


  1. 1.
    Caligiuri MA (2008) Human natural killer cells. Blood 112(3):461–469PubMedCrossRefGoogle Scholar
  2. 2.
    Robertson MJ, Ritz J (1990) Biology and clinical relevance of human natural killer cells. Blood 76:2421–2438PubMedGoogle Scholar
  3. 3.
    Leibson P (1997) Signal transduction during natural killer cell activation: inside the mind of a killer. Immunity 6:655–661PubMedCrossRefGoogle Scholar
  4. 4.
    Trotta R, Fettucciari K, Azzoni L, Abebe B, Puorro KA, Eisenlohr LC, Perussia B (2000) Differential role of p38 and c-Jun N-terminal kinase 1 mitogen-activated protein kinases in NK cell cytotoxicity. J Immunol 165:1782–1789PubMedGoogle Scholar
  5. 5.
    Roda JM, Parihar R, Magro C, Nuovo GJ, Tridandapani S, Carson WE (2006) Natural killer cells produce T cell-recruiting chemokines in response to antibody-coated target cells. Cancer Res 66(1):517–526PubMedCrossRefGoogle Scholar
  6. 6.
    Cartron G, Dacheux L, Salles G, Solal-Celigny P, Bardos P, Colombat P, Watier H (2002) Therapeutic activity of humanized anti-CD20 monoclonal antibody and polymorphism in IgG Fc receptor FcγRIIIa gene. Blood 99:754–758PubMedCrossRefGoogle Scholar
  7. 7.
    Musolino A, Naldi N, Bortesi B, Pezzuolo D, Capelletti M, Missale G, Laccabue D, Zerbini A, Camisa R, Bisagni G, Neri TM, Ardizzoni A (2008) Immunoglobulin G fragment c receptor polymorphisms and clinical efficacy of trastuzumab-based therapy in patients with HER-2/neu-positive metastatic breast cancer. J Clin Oncol 26(11):1789–1796PubMedCrossRefGoogle Scholar
  8. 8.
    Bibeau F, Lopez-Crapez E, Di Fiore F, Thezenas S, Ychou M, Blanchard F, Lamy A, Penault-Llorca F, Frebourg T, Michel P, Sabourin J-C, Boissiere-Michot F (2009) Impact of FcγRIIa-FcγRIIIa polymorphisms and KRAS mutations on the clinical outcome of patients with metastatic colorectal cancer treated with cetuximab plus irinotecan. J Clin Oncol 27(7):1122–1129PubMedCrossRefGoogle Scholar
  9. 9.
    Nakanishi K, Yoshimoto T, Tsutsui H, Okamura H (2001) Interleukin-18 regulates both Th1 and Th2 responses. Ann Rev Immunol 19:423–474CrossRefGoogle Scholar
  10. 10.
    Osaki T, Peron J-M, Cai Q, Okamura H, Robbins PD, Kurimoto M, Lotze MT, Tahara H (1998) IFN-γ-inducing factor/IL-18 administration mediates IFN-γ- and IL-12-independent antitumor effects. J Immunol 160:1742–1749PubMedGoogle Scholar
  11. 11.
    Hashimoto W, Osaki T, Okamura H, Robbins PD, Kurimoto M, Nagata S, Peron J-M, Lotze MT, Tahara H (1999) Differential antitumor effects of administration of recombinant IL-18 or recombinant IL-12 are mediated primarily by Fas–Fas ligand- and perforin-induced tumor apoptosis, respectively. J Immunol 163:583–589PubMedGoogle Scholar
  12. 12.
    Robertson MJ, Mier JW, Logan T, Atkins M, Koon H, Koch KM, Kathman S, Pandite LN, Oei C, Kirby LC, Jewell RC, Bell WN, Thurmond LM, Weisenbach J, Roberts S, Dar MM (2006) Clinical and biological effects of recombinant human interleukin-18 administered by intravenous infusion to patients with advanced cancer. Clin Cancer Res 12(14):4265–4273PubMedCrossRefGoogle Scholar
  13. 13.
    Robertson MJ, Kirkwood JM, Logan TF, Koch KM, Kathman S, Kirby LC, Bell WN, Thurmond LM, Weisenbach J, Dar MM (2008) A dose-escalation study of recombinant human interleukin-18 using two different schedules of administration in patients with cancer. Clin Cancer Res 14(11):3462–3469PubMedCrossRefGoogle Scholar
  14. 14.
    Robertson MJ, Abonour R, Hromas R, Nelson RP, Fineberg NS, Cornetta K (2005) Augmented high-dose regimen of cyclophosphamide, carmustine, and etoposide with autologous hematopoietic stem cell transplantation for relapsed and refractory aggressive non-Hodgkin’s lymphoma. Leuk Lymphoma 46:1477–1487PubMedCrossRefGoogle Scholar
  15. 15.
    Robertson MJ, Caligiuri MA, Manley TJ, Levine H, Ritz J (1990) Human natural killer cell adhesion molecules: differential expression after activation and participation in cytolysis. J Immunol 145:3194–3201PubMedGoogle Scholar
  16. 16.
    Chang H-C, Han L, Goswami R, Nguyen ET, Pelloso D, Robertson MJ, Kaplan MH (2009) Impaired development of human Th1 cells in patients with deficient expression of STAT4. Blood 113(23):5887–5890PubMedCrossRefGoogle Scholar
  17. 17.
    Parihar R, Dierksheide J, Hu Y, Carson WE (2002) IL-12 enhances the natural killer cell cytokine response to Ab-coated target cells. J Clin Invest 110(7):983–992PubMedGoogle Scholar
  18. 18.
    Roda JM, Parihar R, Lehman A, Mani A, Tridandapani S, Carson WE (2006) Interleukin-21 enhances NK cell activation in response to antibody-coated targets. J Immunol 177:120–129PubMedGoogle Scholar
  19. 19.
    Fehniger TA, Shah MH, Turner MJ, VanDeusen JB, Whitman SP, Cooper MA, Suzuki K, Wechser M, Goodsaid F, Caligiuri MA (1999) Differential cytokine and chemokine gene expression by human NK cells following activation with IL-18 or IL-15 in combination with IL-12: implications for the innate immune response. J Immunol 162:4511–4520PubMedGoogle Scholar
  20. 20.
    Robertson MJ, Chang H-C, Pelloso D, Kaplan MH (2005) Impaired interferon-γ production as a consequence of STAT4 deficiency after autologous hematopoietic stem cell transplantation for lymphoma. Blood 106:963–970PubMedCrossRefGoogle Scholar
  21. 21.
    Robertson MJ, Pelloso D, Abonour R, Hromas RA, Nelson RP Jr, Wood L, Cornetta K (2002) Interleukin-12 immunotherapy after autologous stem cell transplantation for hematologic malignancies. Clin Cancer Res 8:3383–3393PubMedGoogle Scholar
  22. 22.
    Trotta R, Dal Col J, Yu J, Ciarlariello D, Thomas B, Zhang X, Allard J, Wei M, Mao H, Byrd JC, Perrotti D, Caligiuri MA (2008) TGF-β utilizes SMAD3 to inhibit CD16-mediated IFN-γ production and antibody-dependent cellular cytotoxicity in human NK cells. J Immunol 181:3784–3792PubMedGoogle Scholar
  23. 23.
    Perussia B, Trinchieri G, Jackson A, Warner NL, Faust J, Rumpold H, Kraft D, Lanier LL (1984) The Fc receptor for IgG on human natural killer cells: phenotypic, functional, and comparative studies with monoclonal antibodies. J Immunol 133(1):180–189PubMedGoogle Scholar
  24. 24.
    Metes D, Ernst LK, Chambers WH, Sulica A, Herberman RB, Morel PA (1998) Expression of functional CD32 molecules on human NK cells is determined by an allelic polymorphism of the FcγRIIC gene. Blood 91(7):2369–2380PubMedGoogle Scholar
  25. 25.
    Metes D, Manciulea M, Petrusca D, Rabinowich H, Ernst LK, Popescu I, Calugaru A, Sulica A, Chambers WH, Herberman RB, Morel PA (1999) Ligand binding specificities and signal transduction pathways of Fcγ receptor IIc isoforms: the CD32 isoforms expressed by human NK cells. Eur J Immunol 29:2842–2852PubMedCrossRefGoogle Scholar
  26. 26.
    Cassatella MA, Anegon I, Cuturi MC, Griskey P, Trinchieri G, Perussia B (1989) Fcg R (CD16) interaction with ligand induces Ca2 + mobilization and phosphoinositide turnover in human natural killer cells: role of Ca2 + in Fcγ R (CD16)-induced transcription and expression of lymphokine genes. J Exp Med 169:549–567PubMedCrossRefGoogle Scholar
  27. 27.
    Mailliard RB, Alber SM, Shen H, Watkins SC, Kirkwood JM, Herberman RB, Kalinski P (2005) IL-18-induced CD83 + CCR7 + NK helper cells. J Exp Med 202(7):941–953PubMedCrossRefGoogle Scholar
  28. 28.
    Schroder K, Hertzog PJ, Ravasi T, Hume DA (2004) Interferon-γ: an overview of signals, mechanisms and functions. J Leukoc Biol 75:163–189PubMedCrossRefGoogle Scholar
  29. 29.
    Kondadasula SV, Roda JM, Parihar R, Yu J, Lehman A, Caligiuri MA, Tridandapani S, Burry RW, Carson WE (2008) Colocalization of the IL-12 receptor and FcγRIIIa to natural killer cell lipid rafts leads to activation of ERK and enhanced production of interferon-g. Blood 111(8):4173–4183PubMedCrossRefGoogle Scholar
  30. 30.
    Mavropoulos A, Sully G, Cope AP, Clark A (2004) Stabilization of IFN-γ mRNA by MAPK p38 in IL-12- and IL-18-stimulated human NK cells. Blood 105:282–288PubMedCrossRefGoogle Scholar
  31. 31.
    Nakahira M, Ahn H-J, Park W-R, Gao P, Tomura M, Park C-S, Hamaoka T, Ohta T, Kurimoto M, Fujiwara H (2002) Synergy of IL-12 and IL-18 for IFN-γ gene expression: IL-12-induced STAT4 contributes to IFN-γ promoter activation by up-regulating the binding activity of IL-18-induced activator protein 1. J Immunol 168:1146–1153PubMedGoogle Scholar
  32. 32.
    Zhang S, Kaplan MH (2000) The p38 mitogen-activated protein kinase is required for IL-12-induced IFN-γ expression. J Immunol 165:1375–1380Google Scholar
  33. 33.
    Robertson MJ (2002) Role of chemokines in the biology of natural killer cells. J Leukoc Biol 71:173–183PubMedGoogle Scholar
  34. 34.
    Overdijk MB, Verploegen S, Buijsse AO, Vink T, Leusen JHW, Bleeker WK, Parren PWHI (2012) Crosstalk between human IgG isotypes and murine effector cells. J Immunol 189:3430–3438PubMedCrossRefGoogle Scholar
  35. 35.
    Daniel D, Yang B, Lawrence DA, Totpal K, Balter I, Lee WP, Gogineni A, Cole MJ, Yee SF, Ross S, Ashkenazi A (2007) Cooperation of the proapoptotic receptor agonist rhApo2L/TRAIL with the CD20 antibody rituximab against non-Hodgkin lymphoma xenografts. Blood 110:4037–4046PubMedCrossRefGoogle Scholar
  36. 36.
    Hernandez-Ilizaliturri FJ, Reddy N, Holkova B, Ottman E, Czuczman MS (2005) Immunomodulatory drug CC-5013 or CC-4047 and rituximab enhance antitumor activity in a severe combined immunodeficient mouse lymphoma model. Clin Cancer Res 11(16):5984–5992PubMedCrossRefGoogle Scholar
  37. 37.
    Wigginton JM, Lee J-K, Wiltrout TA, Alvord WG, Hixon JA, Subleski J, Back TC, Wiltrout RH (2002) Syngergistic enhancement of ineffective endogenous antitumor immune response and induction of IFN-γ and Fas-ligand-dependent tumor eradication by combined administration of IL-18 and IL-2. J Immunol 169:4467–4474PubMedGoogle Scholar
  38. 38.
    Jonak ZL, Trulli S, Maier C, McCabe FL, Kirkpatrick R, Johanson K, Ho YS, Elefante L, Chen Y-J, Herzyk D, Lotze MT, Johnson RK (2002) High-dose recombinant interleukin-18 induces an effective Th1 immune response to murine MOPC-315 plasmacytoma. J Immunother 25(Suppl. 1):S20–S27PubMedCrossRefGoogle Scholar
  39. 39.
    Robertson MJ, Bauman J, Gardner O, Jonak Z, Struemper H, Germaschewski F, Koch KM, Murray S, Weisenbach J, Toso J (2011) A phase I trial evaluating the safety and biological activity of iboctadekin (rhIL-18) in combination with rituximab in patients with CD20 + B cell non-Hodgkin’s lymphoma. Blood 118:1579–1580 (Abstract 3697)CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Shivani Srivastava
    • 1
    • 2
  • David Pelloso
    • 1
    • 2
  • Hailin Feng
    • 1
  • Larry Voiles
    • 3
  • David Lewis
    • 3
  • Zdenka Haskova
    • 4
  • Margaret Whitacre
    • 4
  • Stephen Trulli
    • 4
  • Yi-Jiun Chen
    • 4
  • John Toso
    • 4
  • Zdenka L. Jonak
    • 4
  • Hua-Chen Chang
    • 3
  • Michael J. Robertson
    • 1
    • 2
  1. 1.Bone Marrow and Stem Cell Transplantation ProgramIndianapolisUSA
  2. 2.Lymphoma ProgramIndiana University Medical CenterIndianapolisUSA
  3. 3.Department of BiologyIndiana University-Purdue University School of ScienceIndianapolisUSA
  4. 4.GlaxoSmithKlineKing of PrussiaUSA

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