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Immunological properties and vaccine efficacy of murine dendritic cells simultaneously expressing melanoma-associated antigen and interleukin-12

Abstract

Interleukin (IL)-12 is a key factor for inducing cellular immune responses, which play a central role in the eradication of cancer. In the present study, in order to create a dendritic cell (DC)-based vaccine capable of positively skewing immune response toward a cellular immunity-dominant state, we analyzed immunological characteristics and vaccine efficacy of DCs cotransduced with melanoma-associated antigen (gp100) and IL-12 gene (gp100+IL12/DCs) by using RGD fiber-mutant adenovirus vector (AdRGD), which enables highly efficient gene transduction into DCs. gp100+IL12/DCs could simultaneously express cytoplasmic gp100 and secretory IL-12 at levels comparable to DCs transduced with each gene alone. In comparison with DCs transduced with gp100 alone (gp100/DCs), upregulation of major histocompatibility complex class I, CD40, and CD86 molecules on the cell surface and more potent T-cell-stimulating ability for proliferation and interferon-γ secretion were observed as characteristic changes in gp100+IL12/DCs. In addition, administration of gp100+IL12/DCs, which were prepared by a relatively low dose of AdRGD-IL12, could induce more potent tumor-specific cellular immunity in the murine B16BL6 melanoma model than vaccination with gp100/DCs. However, antitumor effect and B16BL6-specific cytotoxic T-lymphocyte activity in mice vaccinated with gp100+IL12/DCs diminished with increasing AdRGD-IL12 dose during gene transduction, and paralleled the decrease in presentation levels via MHC class I molecules for antigen transduced with another AdRGD. Collectively, our results suggested that optimization of combined vector dose was required for development of a more efficacious DC-based vaccine for cancer immunotherapy, which relied on genetic engineering to simultaneously express tumor-associated antigen and IL-12.

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Abbreviations

2-ME:

2-mercaptoethanol

AdRGD:

RGD fiber-mutant adenovirus vector

APC:

antigen-presenting cell

BrdU:

5-bromo-2′-deoxyuridine

CTL:

cytotoxic T lymphocyte

DC:

dendritic cell

Eu:

europium

FBS:

fetal bovine serum

GM-CSF:

granulocyte/macrophage colony-stimulating factor

IFN:

interferon

IL:

interleukin

LPS:

lipopolysaccharide

mAb:

monoclonal antibody

MHC:

major histocompatibility complex

MLR:

mixed leukocyte reaction

MMC:

mitomycin C

MOI:

multiplicity of infection

NK:

natural killer

OVA:

ovalbumin

PBS:

phosphate-buffered saline

RT-PCR:

reverse transcription-polymerase chain reaction

TAA:

tumor-associated antigen

Th:

helper T cell

References

  1. Banchereau J, Steinman RM . Dendritic cells and the control of immunity. Nature. 1998; 392: 245–252.

    Article  CAS  Google Scholar 

  2. Kapsenberg ML . Dendritic-cell control of pathogen-driven T-cell polarization. Nat Rev Immunol. 2003; 3: 984–993.

    Article  CAS  Google Scholar 

  3. Nestle FO, Alijagic S, Gilliet M, et al. Vaccination of melanoma patients with peptide- or tumor lysate-pulsed dendritic cells. Nat Med. 1998; 4: 328–332.

    Article  CAS  Google Scholar 

  4. Thurner B, Haendle I, Roder C, et al. Vaccination with mage-3A1 peptide-pulsed mature, monocyte-derived dendritic cells expands specific cytotoxic T cells and induces regression of some metastases in advanced stage IV melanoma. J Exp Med. 1999; 190: 1669–1678.

    Article  CAS  Google Scholar 

  5. Yu JS, Wheeler CJ, Zeltzer PM, et al. Vaccination of malignant glioma patients with peptide-pulsed dendritic cells elicits systemic cytotoxicity and intracranial T-cell infiltration. Cancer Res. 2001; 61: 842–847.

    CAS  PubMed  Google Scholar 

  6. Okada N, Tsukada Y, Nakagawa S, et al. Efficient gene delivery into dendritic cells by fiber-mutant adenovirus vectors. Biochem Biophys Res Commun. 2001; 282: 173–179.

    Article  CAS  Google Scholar 

  7. Okada N, Masunaga Y, Okada Y, et al. Gene transduction efficiency and maturation status in mouse bone marrow-derived dendritic cells infected with conventional or RGD fiber-mutant adenovirus vectors. Cancer Gene Ther. 2003; 10: 421–431.

    Article  CAS  Google Scholar 

  8. Okada N, Saito T, Masunaga Y, et al. Efficient antigen gene transduction using Arg-Gly-Asp fiber-mutant adenovirus vectors can potentiate antitumor vaccine efficacy and maturation of murine dendritic cells. Cancer Res. 2001; 61: 7913–7919.

    CAS  PubMed  Google Scholar 

  9. Okada N, Masunaga Y, Okada Y, et al. Dendritic cells transduced with gp100 gene by RGD fiber-mutant adenovirus vectors are highly efficacious in generating anti-B16BL6 melanoma immunity in mice. Gene Therapy. 2003; 10: 1891–1902.

    Article  CAS  Google Scholar 

  10. Hammerling GJ, Klar D, Pulm W, et al. The influence of major histocompatibility complex class I antigens on tumor growth and metastasis. Biochim Biophys Acta. 1987; 907: 245–259.

    CAS  PubMed  Google Scholar 

  11. Moller P, Hammerling GJ . The role of surface HLA-A,B,C molecules in tumour immunity. Cancer Surv. 1992; 13: 101–127.

    CAS  PubMed  Google Scholar 

  12. Khanna R . Tumour surveillance: missing peptides and MHC molecules. Immunol Cell Biol. 1998; 76: 20–26.

    Article  CAS  Google Scholar 

  13. Nishimura T, Nakui M, Sato M, et al. The critical role of Th1-dominant immunity in tumor immunology. Cancer Chemother Pharmacol. 2000; 46 (Suppl):S52–S61.

    Article  CAS  Google Scholar 

  14. Gubler U, Chua AO, Schoenhaut DS, et al. Coexpression of two distinct genes is required to generate secreted bioactive cytotoxic lymphocyte maturation factor. Proc Natl Acad Sci USA. 1991; 88: 4143–4147.

    Article  CAS  Google Scholar 

  15. Wolf SF, Temple PA, Kobayashi M, et al. Cloning of cDNA for natural killer cell stimulatory factor, a heterodimeric cytokine with multiple biologic effects on T and natural killer cells. J Immunol. 1991; 146: 3074–3081.

    CAS  PubMed  Google Scholar 

  16. Robertson MJ, Soiffer RJ, Wolf SF, et al. Response of human natural killer (NK) cells to NK cell stimulatory factor (NKSF): cytolytic activity and proliferation of NK cells are differentially regulated by NKSF. J Exp Med. 1992; 175: 779–788.

    Article  CAS  Google Scholar 

  17. Brunda MJ . Interleukin-12. J Leukoc Biol. 1994; 55: 280–288.

    Article  CAS  Google Scholar 

  18. Chan SH, Perussia B, Gupta JW, et al. Induction of interferon gamma production by natural killer cell stimulatory factor: characterization of the responder cells and synergy with other inducers. J Exp Med. 1991; 173: 869–879.

    Article  CAS  Google Scholar 

  19. Chan SH, Kobayashi M, Santoli D, et al. Mechanisms of IFN-γ induction by natural killer cell stimulatory factor (NKSF/IL-12). Role of transcription and mRNA stability in the synergistic interaction between NKSF and IL-2. J Immunol. 1992; 148: 92–98.

    CAS  PubMed  Google Scholar 

  20. Hsieh CS, Macatonia SE, Tripp CS, et al. Development of TH1 CD4+ T cells through IL-12 produced by Listeria-induced macrophages. Science. 1993; 260: 547–549.

    Article  CAS  Google Scholar 

  21. Seder RA, Gazzinelli R, Sher A, et al. Interleukin 12 acts directly on CD4+ T cells to enhance priming for interferon γ production and diminishes interleukin 4 inhibition of such priming. Proc Natl Acad Sci USA. 1993; 90: 10188–10192.

    Article  CAS  Google Scholar 

  22. Nastala CL, Edington HD, McKinney TG, et al. Recombinant IL-12 administration induces tumor regression in association with IFN-γ production. J Immunol. 1994; 153: 1697–1706.

    CAS  PubMed  Google Scholar 

  23. Voest EE, Kenyon BM, O’Reilly MS, et al. Inhibition of angiogenesis in vivo by interleukin 12. J Natl Cancer Inst. 1995; 87: 581–586.

    Article  CAS  Google Scholar 

  24. Pfeifer JD, Wick MJ, Roberts RL, et al. Phagocytic processing of bacterial antigens for class I MHC presentation to T cells. Nature. 1993; 361: 359–362.

    Article  CAS  Google Scholar 

  25. Mizuguchi H, Koizumi N, Hosono T, et al. A simplified system for constructing recombinant adenoviral vectors containing heterologous peptides in the HI loop of their fiber knob. Gene Therapy. 2001; 8: 730–735.

    Article  CAS  Google Scholar 

  26. Okada Y, Okada N, Nakagawa S, et al. Fiber-mutant technique can augment gene transduction efficacy and anti-tumor effects against established murine melanoma by cytokine-gene therapy using adenovirus vectors. Cancer Lett. 2002; 177: 57–63.

    Article  CAS  Google Scholar 

  27. Mizuguchi H, Kay MA . Efficient construction of a recombinant adenovirus vector by an improved in vitro ligation method. Hum Gene Ther. 1998; 9: 2577–2583.

    Article  CAS  Google Scholar 

  28. Mizuguchi H, Kay MA . A simple method for constructing E1- and E1/E4-deleted recombinant adenoviral vectors. Hum Gene Ther. 1999; 10: 2013–2017.

    Article  CAS  Google Scholar 

  29. Lutz MB, Kukutsch N, Ogilvie AL, et al. An advanced culture method for generating large quantities of highly pure dendritic cells from mouse bone marrow. J Immunol Methods. 1999; 223: 77–92.

    Article  CAS  Google Scholar 

  30. Okada N, Tsujino M, Hagiwara Y, et al. Administration route-dependent vaccine efficiency of murine dendritic cells pulsed with antigens. Br J Cancer. 2001; 84: 1564–1570.

    Article  CAS  Google Scholar 

  31. Lanzavecchia A . Identifying strategies for immune intervention. Science. 1993; 260: 937–944.

    Article  CAS  Google Scholar 

  32. Scott P, Trinchieri G . IL-12 as an adjuvant for cell-mediated immunity. Semin Immunol. 1997; 9: 285–291.

    Article  CAS  Google Scholar 

  33. Shurin MR, Esche C, Peron JM, et al. Antitumor activities of IL-12 and mechanisms of action. Chem Immunol. 1997; 68: 153–174.

    Article  CAS  Google Scholar 

  34. Zitvogel L, Couderc B, Mayordomo JI, et al. IL-12-engineered dendritic cells serve as effective tumor vaccine adjuvants in vivo. Ann NY Acad Sci. 1996; 795: 284–293.

    Article  CAS  Google Scholar 

  35. Nishioka Y, Hirao M, Robbins PD, et al. Induction of systemic and therapeutic antitumor immunity using intratumoral injection of dendritic cells genetically modified to express interleukin 12. Cancer Res. 1999; 59: 4035–4041.

    CAS  PubMed  Google Scholar 

  36. Melero I, Duarte M, Ruiz J, et al. Intratumoral injection of bone-marrow derived dendritic cells engineered to produce interleukin-12 induces complete regression of established murine transplantable colon adenocarcinomas. Gene Therapy. 1999; 6: 1779–1784.

    Article  CAS  Google Scholar 

  37. Okada Y, Okada N, Mizuguchi H, et al. Optimization of antitumor efficacy and safety of in vivo cytokine gene therapy using RGD fiber-mutant adenovirus vector for preexisting murine melanoma. Biochim Biophys Acta. 2004; 1670: 172–180.

    Article  CAS  Google Scholar 

  38. Snijders A, Hilkens CM, van der Pouw Kraan TC, et al. Regulation of bioactive IL-12 production in lipopolysaccharide-stimulated human monocytes is determined by the expression of the p35 subunit. J Immunol. 1996; 156: 1207–1212.

    CAS  PubMed  Google Scholar 

  39. Kalinski P, Vieira PL, Schuitemaker JH, et al. Prostaglandin E2 is a selective inducer of interleukin-12 p40 (IL-12p40) production and an inhibitor of bioactive IL-12p70 heterodimer. Blood. 2001; 97: 3466–3469.

    Article  CAS  Google Scholar 

  40. Koblish HK, Hunter CA, Wysocka M, et al. Immune suppression by recombinant interleukin (rIL)-12 involves interferon gamma induction of nitric oxide synthase 2 (iNOS) activity: inhibitors of NO generation reveal the extent of rIL-12 vaccine adjuvant effect. J Exp Med. 1998; 188: 1603–1610.

    Article  CAS  Google Scholar 

  41. Medot-Pirenne M, Heilman MJ, Saxena M, et al. Augmentation of an antitumor CTL response in vivo by inhibition of suppressor macrophage nitric oxide. J Immunol. 1999; 163: 5877–5882.

    CAS  PubMed  Google Scholar 

  42. Lasarte JJ, Corrales FJ, Casares N, et al. Different doses of adenoviral vector expressing IL-12 enhance or depress the immune response to a coadministered antigen: the role of nitric oxide. J Immunol. 1999; 162: 5270–5277.

    CAS  PubMed  Google Scholar 

  43. Nishioka Y, Wen H, Mitani K, et al. Differential effects of IL-12 on the generation of alloreactive CTL mediated by murine and human dendritic cells: a critical role for nitric oxide. J Leukoc Biol. 2003; 73: 621–629.

    Article  CAS  Google Scholar 

  44. Piccioli D, Sbrana S, Melandri E, et al. Contact-dependent stimulation and inhibition of dendritic cells by natural killer cells. J Exp Med. 2002; 195: 335–341.

    Article  CAS  Google Scholar 

  45. Chen Y, Emtage P, Zhu Q, et al. Induction of ErbB-2/neu-specific protective and therapeutic antitumor immunity using genetically modified dendritic cells: enhanced efficacy by cotransduction of gene encoding IL-12. Gene Therapy. 2001; 8: 316–323.

    Article  CAS  Google Scholar 

  46. Ribas A, Amarnani SN, Buga GM, et al. Immunosuppressive effects of interleukin-12 coexpression in melanoma antigen gene-modified dendritic cell vaccines. Cancer Gene Ther. 2002; 9: 875–883.

    Article  CAS  Google Scholar 

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Acknowledgements

We are grateful to Dr Hiroshi Yamamoto (Department of Immunology, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Japan) for providing mIL12 BIA/pBluescript II KS(-), to Dr Hirofumi Hamada (Department of Molecular Medicine, Sapporo Medical University, Sapporo, Japan) for providing pAx1-CA h-gp100, to Dr Michael J Bevan (Department of Immunology, Howard Hughes Medical Institute, University of Washington, Seattle, WA) for providing pAc-neo-OVA, to Dr Clifford V Harding (Department of Pathology, Case Western Reserve University, Cleveland, OH) for providing CD8-OVA 1.3 cells, to Yasushige Masunaga, Masaya Nishida, and Aya Matsui (Department of Biopharmaceutics, Kyoto Pharmaceutical University, Kyoto, Japan) for technical assistance, and to KIRIN Brewery Co., Ltd (Tokyo, Japan) for providing recombinant murine GM-CSF.

The present study was supported in part by the Research on Health Sciences focusing on Drug Innovation from The Japan Health Sciences Foundation; by the Science Research Promotion Fund of the Japan Private School Promotion Foundation; by grants from the Bioventure Development Program of the Ministry of Education, Culture, Sports, Science and Technology of Japan; and by grants from the Ministry of Health, Labour and Welfare in Japan.

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Correspondence to Naoki Okada.

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Okada, N., Iiyama, S., Okada, Y. et al. Immunological properties and vaccine efficacy of murine dendritic cells simultaneously expressing melanoma-associated antigen and interleukin-12. Cancer Gene Ther 12, 72–83 (2005). https://doi.org/10.1038/sj.cgt.7700772

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