Dendritic cells combined with doxorubicin induces immunogenic cell death and exhibits antitumor effects for osteosarcoma

Kawano,Masanori; Tanaka,Kazuhiro; Itonaga,Ichiro; Iwasaki,Tatsuya; Miyazaki,Masashi; Ikeda,Shinichi; Tsumura,Hiroshi

doi:10.3892/ol.2016.4175

Dendritic cells combined with doxorubicin induces immunogenic cell death and exhibits antitumor effects for osteosarcoma

Authors:
- Masanori Kawano
- Kazuhiro Tanaka
- Ichiro Itonaga
- Tatsuya Iwasaki
- Masashi Miyazaki
- Shinichi Ikeda
- Hiroshi Tsumura
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Affiliations: Department of Orthopaedic Surgery, Faculty of Medicine, Oita University, Yufu, Oita 879‑5593, Japan
Published online on: February 2, 2016 https://doi.org/10.3892/ol.2016.4175
Pages: 2169-2175

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Abstract

The effects of dendritic cells (DCs) with low dose doxorubicin on the enhancement of the systemic immune response, including the effects on calreticulin (CRT) expression, heat shock protein 70 (HSP70) on the cell surface expression, and the enhancement of high mobility group box 1 (HMGB1) release from cancer cells, remain unclear. The present study investigated whether the combination of DCs and doxorubicin (ADM) induces immune cell death, and leads to tumor growth inhibition in a murine osteosarcoma model. To evaluate immune response activation in vivo, 4 groups of mice were established: i) untreated mice, ii) DC‑treated mice, iii) ADM‑treated mice, and iv) DC and ADM‑treated mice. Immunological cell death and CRT, HSP70, and HMGB1 expression levels were higher in doxorubicin-treated cells than those in untreated or those treated with DCs alone. NF-κB expression was higher in the DCs after ligand activation using CRT, HSP70, or HMGB1 in vitro. Mice treated with DCs and ADM displayed an increased number of CD8+ T‑lymphocytes within metastatic tumors and inhibition of metastatic growth. The expression of CRT and the release of HMGB1 from tumor tissues were increased in the ADM‑treated groups. Treatment with DCs and ADM resulted in the highest serum interferon‑γ levels. Combining ADM, which can induce immunogenic cell death, with DCs enhanced the systemic immune response. The findings of the present study provide further support for the continued development of antitumor agents that induce cell death and the immune response to target osteosarcoma.

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1	Stiller CA, Bielack SS, Jundt G and Steliarova-Foucher E: Bone tumours in European children and adolescents, 1978–1997. Report from the Automated Childhood Cancer Information System project. Eur J Cancer. 42:2124–2135. 2006. View Article : Google Scholar : PubMed/NCBI
2	Bielack SS, Kempf-Bielack B, Delling G, Exner GU, Flege S, Helmke K, Kotz R, Salzer-Kuntschik M, Werner M, Winkelmann W, et al: Prognostic factors in high-grade osteosarcoma of the extremities or trunk: An analysis of 1,702 patients treated on neoadjuvant cooperative osteosarcoma study group protocols. J Clin Oncol. 20:776–790. 2002. View Article : Google Scholar : PubMed/NCBI
3	Milosević DB: The different level of immunological recovery after chemotherapy in leukemia and lymphoma patients. J Exp Clin Cancer Res. 20:517–522. 2001.PubMed/NCBI
4	Tesniere A, Panaretakis T, Kepp O, Apetoh L, Ghiringhelli F, Zitvogel L and Kroemer G: Molecular characteristics of immunogenic cancer cell death. Cell Death Differ. 15:3–12. 2008. View Article : Google Scholar : PubMed/NCBI
5	Apetoh L, Mignot G, Panaretakis T, Kroemer G and Zitvogel L: Immunogenicity of anthracyclines: Moving towards more personalized medicine. Trends Mol Med. 14:141–151. 2008. View Article : Google Scholar : PubMed/NCBI
6	Apetoh L, Ghiringhelli F, Tesniere A, Obeid M, Ortiz C, Criollo A, Mignot G, Maiuri MC, Ullrich E, Saulnier P, et al: Toll-like receptor 4-dependent contribution of the immune system to anticancer chemotherapy and radiotherapy. Nature. 13:1050–1059. 2007.
7	Fucikova J, Kralikova P, Fialova A, Brtnicky T, Rob L, Bartunkova J and Spísek R: Human tumor cells killed by anthracyclines induce a tumor-specific immune response. Cancer Res. 71:4821–4833. 2011. View Article : Google Scholar : PubMed/NCBI
8	Sancho D, Joffre OP, Keller AM, Rogers NC, Martínez D, Hernanz-Falcón P, Rosewell I and Sousa Reise C: Identification of a dendritic cell receptor that couples sensing of necrosis to immunity. Nature. 458:899–903. 2009. View Article : Google Scholar : PubMed/NCBI
9	Lutz MB and Rössner S: Factors influencing the generation of murine dendritic cells from bone marrow: The special role of fetal calf serum. Immunobiology. 212:855–862. 2007. View Article : Google Scholar : PubMed/NCBI
10	Kawano M, Nishida H, Nakamoto Y, Tsumura H and Tsuchiya H: Cryoimmunologic antitumor effects enhanced by dendritic cells in osteosarcoma. Clin Orthop Relat Res. 468:1373–1383. 2010. View Article : Google Scholar : PubMed/NCBI
11	Kawano M, Itonaga I, Iwasaki T, Tsuchiya H and Tsumura H: Anti-TGF-β antibody combined with dendritic cells produce antitumor effects in osteosarcoma. Clin Orthop Relat Res. 470:2288–2294. 2012. View Article : Google Scholar : PubMed/NCBI
12	Kawano M, Itonaga I, Iwasaki T and Tsumura H: Enhancement of antitumor immunity by combining anti-cytotoxic T lymphocyte antigen-4 antibodies and cryotreated tumor lysate-pulsed dendritic cells in murine osteosarcoma. Oncol Rep. 29:1001–1006. 2013.PubMed/NCBI
13	Zitvogel L, Kepp O and Kroemer G: Immune parameters affecting the efficacy of chemotherapeutic regimens. Nat Rev Clin Oncol. 8:151–160. 2011. View Article : Google Scholar : PubMed/NCBI
14	Locher C, Conforti R, Aymeric L, Ma Y, Yamazaki T, Rusakiewicz S, Tesnière A, Ghiringhelli F, Apetoh L, Morel Y, et al: Desirable cell death during anticancer chemotherapy. Ann N Y Acad Sci. 1209:99–108. 2010. View Article : Google Scholar : PubMed/NCBI
15	Zitvogel L, Kepp O, Senovilla L, Menger L, Chaput N and Kroemer G: Immunogenic tumor cell death for optimal anticancer therapy: The calreticulin exposure pathway. Clin Cancer Res. 16:3100–3104. 2010. View Article : Google Scholar : PubMed/NCBI
16	Obeid M, Tesniere A, Ghiringhelli F, Fimia GM, Apetoh L, Perfettini JL, Castedo M, Mignot G, Panaretakis T, Casares N, et al: Calreticulin exposure dictates the immunogenicity of cancer cell death. Nat Med. 13:54–61. 2007. View Article : Google Scholar : PubMed/NCBI
17	Michaud M, Sukkurwala AQ, Di Sano F, Zitvogel L, Kepp O and Kroemer G: Synthetic induction of immunogenic cell death by genetic stimulation of endoplasmic reticulum stress. Oncoimmunology. 3:e282762014. View Article : Google Scholar : PubMed/NCBI
18	Hoshino K and Kaisho T: Nucleic acid sensing toll-like receptors in dendritic cells. Curr Opin Immunol. 20:408–413. 2008. View Article : Google Scholar : PubMed/NCBI
19	Wang H, Vishnubhakat JM, Bloom O, Zhang M, Ombrellino M, Sama A and Tracey KJ: Proinflammatory cytokines (tumor necrosis factor and interleukin 1) stimulate release of high mobility group protein-1 by pituicytes. Surgery. 126:389–392. 1999. View Article : Google Scholar : PubMed/NCBI
20	Scaffidi P, Misteli T and Bianchi ME: Release of chromatin protein HMGB1 by necrotic cells triggers inflammation. Nature. 418:191–195. 2002. View Article : Google Scholar : PubMed/NCBI
21	Park JS, Svetkauskaite D, He Q, Kim JY, Strassheim D, Ishizaka A and Abraham E: Involvement of toll-like receptors 2 and 4 in cellular activation by high mobility group box 1 protein. J Biol Chem. 279:7370–7377. 2004. View Article : Google Scholar : PubMed/NCBI
22	Apetoh L, Ghiringhelli F, Tesniere A, Criollo A, Ortiz C, Lidereau R, Mariette C, Chaput N, Mira JP, Delaloge S, et al: The interaction between HMGB1 and TLR4 dictates the outcome of anticancer chemotherapy and radiotherapy. Immunol Rev. 220:47–59. 2007. View Article : Google Scholar : PubMed/NCBI
23	Rosenberg SA, Restifo NP, Yang JC, Morgan RA and Dudley ME: Adoptive cell transfer: A clinical path to effective cancer immunotherapy. Nat Rev Cancer. 8:299–308. 2008. View Article : Google Scholar : PubMed/NCBI
24	Nowak AK, Lake RA and Robinson BW: Combined chemoimmunotherapy of solid tumours: Improving vaccines? Adv Drug Deliv Rev. 58:975–990. 2006. View Article : Google Scholar : PubMed/NCBI