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
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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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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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DOI: https://doi.org/10.1038/sj.cgt.7700772
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