Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

Human fused NKG2D–IL-15 protein controls xenografted human gastric cancer through the recruitment and activation of NK cells

Cellular & Molecular Immunology volume 14, pages 293–307 (2017)Cite this article

Abstract

Interleukin (IL)-15 plays an important role in natural killer (NK) and CD8+ T-cell proliferation and function and is more effective than IL-2 for tumor immunotherapy. The trans-presentation of IL-15 by neighboring cells is more effective for NK cell activation than its soluble IL-15. In this study, the fusion protein dsNKG2D–IL-15, which consisted of two identical extracellular domains of human NKG2D coupled to human IL-15 via a linker, was engineered in Escherichia coli. DsNKG2D–IL-15 could efficiently bind to major histocompatibility complex class I chain-related protein A (MICA) of human tumor cells with the two NKG2D domains and trans-present IL-15 to NK or CD8+ T cells. We transplanted human gastric cancer (SGC-7901) cells into nude mice and mouse melanoma cells with ectopic expression of MICA (B16BL6–MICA) into C57BL/6 mice. Then, we studied the anti-tumor effects mediated by dsNKG2D–IL-15 in the two xenografted tumor models. Human dsNKG2D–IL-15 exhibited higher efficiency than IL-15 in suppressing gastric cancer growth. Exogenous human dsNKG2D–IL-15 was centrally distributed in the mouse tumor tissues based on in vivo live imaging. The frequencies of human CD56+ cells infiltrated into the tumor tissues following the injection of peripheral blood mononuclear cells into nude mice bearing human gastric cancer were significantly increased by human dsNKG2D–IL-15 treatment. Human dsNKG2D–IL-15 also delayed the growth of transplanted melanoma (B16BL6–MICA) by activating and recruiting mouse NK and CD8+ T cells. The anti-melanoma effect of human dsNKG2D–IL-15 in C57BL/6 mice was mostly decreased by the in vivo depletion of mouse NK cells. These data highlight the potential use of human dsNKG2D–IL-15 for tumor therapy.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 2 (continued)
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  1. Steel JC, Waldmann TA, Morris JC . Interleukin-15 biology and its therapeutic implications in cancer. Trends Pharmacol Sci 2012; 33: 35–41.

    Article  CAS  Google Scholar 

  2. Jakobisiak M, Golab J, Lasek W . Interleukin 15 as a promising candidate for tumor immunotherapy. Cytokine Growth Factor Rev 2011; 22: 99–108.

    Article  CAS  Google Scholar 

  3. Sanjabi S, Mosaheb MM, Flavell RA . Opposing effects of TGF-beta and IL-15 cytokines control the number of short-lived effector CD8+ T cells. Immunity 2009; 31: 131–144.

    Article  CAS  Google Scholar 

  4. Tao Q, Chen T, Tao L, Wang H, Pan Y, Xiong S et al. IL-15 improves the cytotoxicity of cytokine-induced killer cells against leukemia cells by upregulating CD3+CD56+ cells and downregulating regulatory T cells as well as IL-35. J Immunother 2013; 36: 462–467.

    Article  CAS  Google Scholar 

  5. Berger C, Berger M, Hackman RC, Gough M, Elliott C, Jensen MC et al. Safety and immunologic effects of IL-15 administration in nonhuman primates. Blood 2009; 114: 2417–2426.

    Article  CAS  Google Scholar 

  6. Imamura M, Shook D, Kamiya T, Shimasaki N, Chai SM, Coustan-Smith E et al. Autonomous growth and increased cytotoxicity of natural killer cells expressing membrane-bound interleukin-15. Blood 2014; 124: 1081–1088.

    Article  CAS  Google Scholar 

  7. Yiang GT, Chou RH, Chang WJ, Wei CW, Yu YL . Long-term expression of rAAV2-hIL15 enhances immunoglobulin production and lymphokine-activated killer cell-mediated human glioblastoma cell death. Mol Clin Oncol 2013; 1: 321–325.

    Article  Google Scholar 

  8. Zhao DX, Li ZJ, Zhang Y, Zhang XN, Zhao KC, Li YG et al. Enhanced antitumor immunity is elicited by adenovirus-mediated gene transfer of CCL21 and IL-15 in murine colon carcinomas. Cell Immunol 2014; 289: 155–161.

    Article  CAS  Google Scholar 

  9. Bessard A, Solé V, Bouchaud G, Quéméner A, Jacques Y . High antitumor activity of RLI, an interleukin-15 (IL-15)-IL-15 receptor alpha fusion protein, in metastatic melanoma and colorectal cancer. Mol Cancer Ther 2009; 8: 2736–2745.

    Article  CAS  Google Scholar 

  10. Kermer V, Baum V, Hornig N, Kontermann RE, Müller D . An antibody fusion protein for cancer immunotherapy mimicking IL-15 trans-presentation at the tumor site. Mol Cancer Ther 2012; 11: 1279–1288.

    Article  CAS  Google Scholar 

  11. Cheever MA . Twelve immunotherapy drugs that could cure cancers. Immunol Rev 2008; 222: 357–368.

    Article  CAS  Google Scholar 

  12. Müller JR, Waldmann TA, Kruhlak MJ, Dubois S . Paracrine and trans-presentation functions of IL-15 are mediated by diverse splice versions of IL-15Rα in human monocytes and dendritic cells. J Biol Chem 2012; 287: 40328–40338.

    Article  Google Scholar 

  13. Lucas M, Schachterle W, Oberle K, Aichele P, Diefenbach A . Dendritic cells prime natural killer cells by trans-presenting interleukin 15. Immunity 2007; 26: 503–517.

    Article  CAS  Google Scholar 

  14. Castillo EF, Schluns KS . Regulating the immune system via IL-15 trans-presentation. Cytokine 2012; 59: 479–490.

    Article  CAS  Google Scholar 

  15. Huntington ND, Alves NL, Legrand N, Lim A, Strick-Marchand H, Mention JJ et al. IL-15 trans-presentation promotes both human T-cell reconstitution and T-cell-dependent antibody responses in vivo. Proc Natl Acad Sci USA 2011; 108: 6217–6222.

    Article  CAS  Google Scholar 

  16. Kobayashi H, Dubois S, Sato N, Sabzevari H, Sakai Y, Waldmann TA et al. Role of trans-cellular IL-15 presentation in the activation of NK cell-mediated killing, which leads to enhanced tumor immunosurveillance. Blood 2005; 105: 721–727.

    Article  CAS  Google Scholar 

  17. Stoklasek TA, Schluns KS, Lefrançois L . Combined IL-15/IL-15Ralpha immunotherapy maximizes IL-15 activity in vivo. J Immunol 2006; 177: 6072–6080.

    Article  CAS  Google Scholar 

  18. Dubois S, Patel HJ, Zhang M, Waldmann TA, Müller JR . Preassociation of IL-15 with IL-15R alpha-IgG1-Fc enhances its activity on proliferation of NK and CD8+/CD44high T cells and its antitumor action. J Immunol 2008; 180: 2099–2106.

    Article  CAS  Google Scholar 

  19. Waldmann TA, Conlon KC, Stewart DM, Worthy TA, Janik JE, Fleisher TA et al. Phase 1 trial of IL-15 trans presentation blockade using humanized Mikβ1 mAb in patients with T-cell large granular lymphocytic leukemia. Blood 2013; 121: 476–484.

    Article  CAS  Google Scholar 

  20. Bouchaud G, Garrigue-Antar L, Solé V, Quéméner A, Boublik Y, Mortier E et al. The exon-3-encoded domain of IL-15R alpha contributes to IL-15 high-affinity binding and is crucial for the IL-15 antagonistic effect of soluble IL-15Ralpha. J Mol Biol 2008; 382: 1–12.

    Article  CAS  Google Scholar 

  21. Ullrich E, Koch J, Cerwenka A, Steinle A . New prospects on the NKG2D/NKG2DL system for oncology. Oncoimmunology 2013; 2: e26097 .

    Article  Google Scholar 

  22. Wu J, Song Y, Bakker AB, Bauer S, Spies T, Lanier LL et al. An activating immunoreceptor complex formed by NKG2D and DAP10. Science 1999; 285: 730–732.

    Article  CAS  Google Scholar 

  23. Hayakawa Y . Targeting NKG2D in tumor surveillance. Expert Opin Ther Targets 2012; 16: 587–599.

    Article  CAS  Google Scholar 

  24. Salih HR, Holdenrieder S, Steinle A . Soluble NKG2D ligands: prevalence, release, and functional impact. Front Biosci 2008; 13: 3448–3456.

    Article  CAS  Google Scholar 

  25. Zhang T, Sentman CL . Cancer immunotherapy using a bispecific NK receptor fusion protein that engages both T cells and tumor cells. Cancer Res 2011; 71: 2066–2076.

    Article  CAS  Google Scholar 

  26. Xia Y, Chen B, Shao X, Xiao W, Qian L, Ding Y et al. Treatment with a fusion protein of the extracellular domains of NKG2D to IL-15 retards colon cancer growth in mice. J Immunother 2014; 37: 257–266.

    Article  CAS  Google Scholar 

  27. Gong W, Xiao W, Hu M, Weng X, Qian L, Pan X et al. Ex vivo expansion of natural killer cells with high cytotoxicity by K562 cells modified to co-express major histocompatibility complex class I chain-related protein A, 4-1BB ligand, and interleukin-15. Tissue Antigens 2010; 76: 467–475.

    Article  CAS  Google Scholar 

  28. Lin Z, Wang C, Xia H, Liu W, Xiao W, Qian L et al. CD4(+) NKG2D(+) T cells induce NKG2D down-regulation in natural killer cells in CD86-RAE-1ε transgenic mice. Immunology 2014; 141: 401–415.

    Article  CAS  Google Scholar 

  29. Mrózek E, Anderson P, Caligiuri MA . Role of interleukin-15 in the development of human CD56+ natural killer cells from CD34+ hematopoietic progenitor cells. Blood 1996; 87: 2632–2640.

    PubMed  Google Scholar 

  30. Barao I, Alvarez M, Redelman D, Weiss JM, Ortaldo JR, Wiltrout RH et al. Hydrodynamic delivery of human IL-15 cDNA increases murine natural killer cell recovery after syngeneic bone marrow transplantation. Biol Blood Marrow Transplant 2011; 17: 1754–1764.

    Article  CAS  Google Scholar 

  31. Bauer S, Groh V, Wu J, Steinle A, Phillips JH, Lanier LL et al. Activation of NK cells and T cells by NKG2D, a receptor for stress-inducible MICA. Science 1999; 285: 727–729.

    Article  CAS  Google Scholar 

  32. Cerwenka A, Baron JL, Lanier LL . Ectopic expression of retinoic acid early inducible-1 gene (RAE-1) permits natural killer cell-mediated rejection of a MHC class I-bearing tumor in vivo. Proc Natl Acad Sci USA 2001; 98: 11521–11526.

    Article  CAS  Google Scholar 

  33. Diefenbach A, Jensen ER, Jamieson AM, Raulet DH . Rae1 and H60 ligands of the NKG2D receptor stimulate tumour immunity. Nature 2001; 413: 165–171.

    Article  CAS  Google Scholar 

  34. Zhang J, Xu Z, Zhou X, Zhang H, Yang N, Wu Y et al. Loss of expression of MHC class I-related chain A (MICA) is a frequent event and predicts poor survival in patients with hepatocellular carcinoma. Int J Clin Exp Pathol 2014; 7: 3123–3231.

    PubMed  PubMed Central  Google Scholar 

  35. Chitadze G, Lettau M, Bhat J, Wesch D, Steinle A, Fürst D et al. Shedding of endogenous MHC class I-related chain molecules A and B from different human tumor entities: heterogeneous involvement of the “a disintegrin and metalloproteases” 10 and 17. Int J Cancer 2013; 133: 1557–1566.

    Article  CAS  Google Scholar 

  36. Kaiser BK, Yim D, Chow IT, Gonzalez S, Dai Z, Mann HH et al. Disulphide-isomerase-enabled shedding of tumor-associated NKG2D ligands. Nature 2007; 447: 482–486.

    Article  CAS  Google Scholar 

  37. Chitadze G, Bhat J, Lettau M, Janssen O, Kabelitz D . Generation of soluble NKG2D ligands: proteolytic cleavage, exosome secretion and functional implications. Scand J Immunol 2013; 78: 120–129.

    Article  CAS  Google Scholar 

  38. Dai Z, Turtle CJ, Booth GC, Riddell SR, Gooley TA, Stevens AM et al. Normally occurring NKG2D+CD4+ T cells are immunosuppressive and inversely correlated with disease activity in juvenile-onset lupus. J Exp Med 2009; 206: 793–805.

    Article  CAS  Google Scholar 

  39. Groh V, Smythe K, Dai Z, Spies T . Fas-ligand-mediated paracrine T cell regulation by the receptor NKG2D in tumor immunity. Nat Immunol 2006; 7: 755–762.

    Article  CAS  Google Scholar 

  40. Zhang C, Zhang J, Niu J, Zhang J, Tian Z . Interleukin-15 improves cytotoxicity of natural killer cells via up-regulating NKG2D and cytotoxic effector molecule expression as well as STAT1 and ERK1/2 phosphorylation. Cytokine 2008; 42: 128–136.

    Article  CAS  Google Scholar 

  41. Soriani A, Zingoni A, Cerboni C, Iannitto ML, Ricciardi MR, Di Gialleonardo V et al. ATM-ATR-dependent up-regulation of DNAM-1 and NKG2D ligands on multiple myeloma cells by therapeutic agents results in enhanced NK-cell susceptibility and is associated with a senescent phenotype. Blood 2009; 113: 3503–3511.

    Article  CAS  Google Scholar 

  42. Diermayr S, Himmelreich H, Durovic B, Mathys-Schneeberger A, Siegler U, Langenkamp U et al. NKG2D ligand expression in AML increases in response to HDAC inhibitor valproic acid and contributes to allorecognition by NK-cell lines with single KIR-HLA class I specificities. Blood 2008; 111: 1428–1436.

    Article  CAS  Google Scholar 

  43. Bouchaud G, Gehrke S, Krieg C, Kolios A, Hafner J, Navarini AA et al. Epidermal IL-15Rα acts as an endogenous antagonist of psoriasiform inflammation in mouse and man. J Exp Med 2013; 210: 2105–2117.

    Article  CAS  Google Scholar 

  44. Wendel M, Galani IE, Suri-Payer E, Cerwenka A . Natural killer cell accumulation in tumors is dependent on IFN-gamma and CXCR3 ligands. Cancer Res 2008; 68: 8437–8445.

    Article  CAS  Google Scholar 

  45. González S, Groh V, Spies T . Immunobiology of human NKG2D and its ligands. Curr Top Microbiol Immunol 2006; 298: 121–138.

    PubMed  Google Scholar 

  46. Giri JG, Kumaki S Ahdieh M Friend DJ, Loomis A, Shanebeck K et al. Identification and cloning of a novel IL-15 binding protein that is structurally related to the alpha chain of the IL-2 receptor. EMBO J 1995; 14: 3654–3663.

    Article  CAS  Google Scholar 

  47. Raulet DH, Gasser S, Gowen BG, Deng W, Jung H . Regulation of ligands for the NKG2D activating receptor. Annu Rev Immunol 2013; 31: 413–441.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by the National Natural Science Foundation (nos. 81172785 and 81471547), the Natural Science Fund of Jiangsu Province (BK2011449 and BK2008215), and the collaborative fund of Yangzhou University and Yangzhou municipality (2012038-5).

Author information

Author notes

  1. Yan Chen, Bei Chen and Ti Yang: These authors contributed equally to this article.

  2. W Gong, 1Department of Immunology, School of Medicine, and 2Department of Gastroenterology, The Second Affiliated Hospital, and 3Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, and 4Jiangsu Key Laboratory of Zoonosis, Yangzhou University, and 5Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, 225009, People's Republic of China. E-mail: wjgong@yzu.edu.cn

Authors and Affiliations

  1. Department of Immunology, School of Medicine, Yangzhou University, Yangzhou, 225001, People’s Republic of China

    Yan Chen, Ti Yang, Li Qian, Mingchun Ji & Weijuan Gong

  2. Department of Pharmacology, School of Medicine, Yangzhou University, Yangzhou, 225001, People’s Republic of China

    Bei Chen & Xiaoqun Ge

  3. Department of Gastroenterology, The Second Affiliated Hospital, Yangzhou University, Yangzhou, 225001, People’s Republic of China

    Weiming Xiao, Yanbing Ding & Weijuan Gong

  4. Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for Prevention and Treatment of Senile Diseases, Yangzhou University, Yangzhou, 225009, People’s Republic of China

    Weijuan Gong

  5. Jiangsu Key Laboratory of Zoonosis, Yangzhou University, Yangzhou, 225009, People’s Republic of China

    Weijuan Gong

  6. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou, 225009, People’s Republic of China

    Weijuan Gong

Authors
  1. Yan Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Bei Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ti Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Weiming Xiao
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Li Qian
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yanbing Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Mingchun Ji
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Xiaoqun Ge
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Weijuan Gong
    View author publications

    You can also search for this author in PubMed Google Scholar

Ethics declarations

Competing interests

All authors have declared there are no financial conflicts of interest with regard to this work.

Additional information

Supplementary information of this article can be found on the Cellular & Molecular Immunology's website (http://www.nature.com/cmi).

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chen, Y., Chen, B., Yang, T. et al. Human fused NKG2D–IL-15 protein controls xenografted human gastric cancer through the recruitment and activation of NK cells. Cell Mol Immunol 14, 293–307 (2017). https://doi.org/10.1038/cmi.2015.81

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/cmi.2015.81

Keywords

This article is cited by

Search

Advanced search

Quick links