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Oral administration of poly-gamma-glutamate induces TLR4- and dendritic cell-dependent antitumor effect

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Abstract

Previously, we reported that the oral administration of high molecular mass poly-γ-glutamate (γ-PGA) induced antitumor immunity but the mechanism underlying this antitumor activity was not understood. In the present study, we found that application of high molecular mass γ-PGA induced secretion of tumor necrosis factor (TNF)-α from the bone-marrow-derived macrophages of wild type (C57BL/6 and C3H/HeN) and Toll-like receptor 2 knockout (TLR2−/−) mice, but not those of myeloid differentiation factor 88 knockout (MyD88−/−) and TLR4-defective mice (C3H/HeJ). Production of interferon (IFN)-γ-inducible protein 10 (IP-10) in response to treatment with γ-PGA was almost abolished in C3H/HeJ mice. In contrast to LPS, γ-PGA induced productions of TNF-α and IP-10 could not be blocked by polymyxin B. Furthermore, γ-PGA-induced interleukin-12 production was also impaired in immature dendritic cells (iDCs) from MyD88−/− and C3H/HeJ mice. Downregulation of MyD88 and TLR4 expression using small interfering RNA (siRNA) significantly inhibited γ-PGA-induced TNF-α secretion from the RAW264.7 cells. γ-PGA-mediated intracellular signaling was markedly inhibited in C3H/HeJ cells. The antitumor effect of γ-PGA was completely abrogated in C3H/HeJ mice compared with control mice (C3H/HeN) but significant antitumor effect was generated by the intratumoral administration of C3H/HeN mice-derived iDCs followed by 2,000 kDa γ-PGA in C3H/HeJ. These findings strongly suggest that the antitumor activity of γ-PGA is mediated by TLR4.

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Abbreviations

γ-PGA:

Poly-gamma-glutamate

MyD88:

Myeloid differentiation factor 88

TLR4:

Toll-like receptor 4

JNK:

c-Jun N-terminal kinase

IRF-3:

Interferon regulatory factor-3

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Acknowledgments

We thank Dr S. Akira and Dr S. Uemastu for MyD88−/− and TLR2−/− mice, Dr J. Y. Lee and Dr Kate Fitzgerald for IRF3 luciferase plasmid, and Dr H. Kuroda for helpful advices. This work was supported by grants of the Korea Health 21 R&D Project (A050562) and National R&D Program for Cancer Control (0720510), Ministry of Health & Welfare, Republic of Korea and a grant from KRIBB Initiative program to H. Poo.

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Authors and Affiliations

  1. Mucosal Immunology Laboratory, BioNanotechnology Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 305-806, Korea

    Tae-Young Lee, Yang-Hyun Kim, Sun-Woo Yoon & Haryoung Poo

  2. BioLeaders Corporation, Daejeon, Korea

    Jai-Chul Choi & Moon-Hee Sung

  3. Department of Life Science, Sogang University, Seoul, Korea

    Yang-Hyun Kim & Jai-Myung Yang

  4. College of Veterinary Medicine, Chungnam National University, Daejeon, Korea

    Chul-Joong Kim

  5. Laboratory of Cellular Oncology National Cancer Institute, National Institute of Health, Bethesda, MD, 20892, USA

    John T. Schiller

  6. Department of Bio and Nanochemistry, Kookmin University, Seoul, Korea

    Moon-Hee Sung

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  1. Tae-Young Lee
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Correspondence to Haryoung Poo.

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Lee, TY., Kim, YH., Yoon, SW. et al. Oral administration of poly-gamma-glutamate induces TLR4- and dendritic cell-dependent antitumor effect. Cancer Immunol Immunother 58, 1781–1794 (2009). https://doi.org/10.1007/s00262-009-0689-4

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  • DOI: https://doi.org/10.1007/s00262-009-0689-4

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