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Rejection of intradermally injected syngeneic tumor cells from mice by specific elimination of tumor-associated macrophages with liposome-encapsulated dichloromethylene diphosphonate, followed by induction of CD11b+/CCR3−/Gr-1− cells cytotoxic against the tumor cells

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Abstract

Tumor cell expansion relies on nutrient supply, and oxygen limitation is central in controlling neovascularization and tumor spread. Monocytes infiltrate into tumors from the circulation along defined chemotactic gradients, differentiate into tumor-associated macrophages (TAMs), and then accumulate in the hypoxic areas. Elevated TAM density in some regions or overall TAM numbers are correlated with increased tumor angiogenesis and a reduced host survival in the case of various types of tumors. To evaluate the role of TAMs in tumor growth, we here specifically eliminated TAMs by in vivo application of dichloromethylene diphosphonate (DMDP)-containing liposomes to mice bearing various types of tumors (e.g., B16 melanoma, KLN205 squamous cell carcinoma, and 3LL Lewis lung cancer), all of which grew in the dermis of syngeneic mouse skin. When DMDP-liposomes were injected into four spots to surround the tumor on day 0 or 5 after tumor injection and every third day thereafter, both the induction of TAMs and the tumor growth were suppressed in a dose-dependent and injection number-dependent manner; and unexpectedly, the tumor cells were rejected by 12 injections of three times-diluted DMDP-liposomes. The absence of TAMs in turn induced the invasion of inflammatory cells into or around the tumors; and the major population of effector cells cytotoxic against the target tumor cells were CD11b+ monocytic macrophages, but not CCR3+ eosinophils or Gr-1+ neutrophils. These results indicate that both the absence of TAMs and invasion of CD11b+ monocytic macrophages resulted in the tumor rejection.

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Abbreviations

AIM:

Allograft-induced macrophage

DMDP:

Dichloromethylene diphosphonate

E/T:

Effector/target

FACS:

Fluorescence-activated cell sorter

FCS:

Fetal calf serum

i.d.:

Intradermally

mAb:

Monoclonal antibody

PBS:

Phosphate-buffered saline

PE:

Phycoerythrin

TAMs:

Tumor-associated macrophages

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Acknowledgments

We thank T. Ueno and Y. Fujioka for their skillful technical assistance. This work was supported in part by the Mori and Magari Memorial Research Funds from Osaka Medical College and by a grant-in-aid for scientific research from the Ministry of Education, Science, and Culture, Japan.

Conflict of interest statement

The authors have no financial conflict of interest.

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Authors and Affiliations

  1. Department of Physiology, Osaka Medical College, 2-7 Daigaku-machi, Takatsuki, 569-8686, Japan

    Takeshi Takahashi, Minenori Ibata, Junko Tashiro-Yamaji, Sayako Miura-Takeda, Tetsunosuke Shimizu, Takahiro Kubota & Ryotaro Yoshida

  2. Department of Plastic and Reconstructive Surgery, Osaka Medical College, Takatsuki, 569-8686, Japan

    Takeshi Takahashi, Masashi Okada & Koichi Ueda

  3. Department of Molecular Regulation, Graduate School of Dentistry, Tohoku University, Sendai, Miyagi, 980-8575, Japan

    Zhiqian Yu, Yosuke Shikama & Yasuo Endo

  4. Department of Oral Anatomy and Developmental Biology, Showa University School of Dentistry, 1-5-8 Hatanodai, Shinagawa, Tokyo, 142-8555, Japan

    Yasunori Miyauchi & Masanori Nakamura

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  1. Takeshi Takahashi
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Correspondence to Ryotaro Yoshida.

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Takahashi, T., Ibata, M., Yu, Z. et al. Rejection of intradermally injected syngeneic tumor cells from mice by specific elimination of tumor-associated macrophages with liposome-encapsulated dichloromethylene diphosphonate, followed by induction of CD11b+/CCR3−/Gr-1− cells cytotoxic against the tumor cells. Cancer Immunol Immunother 58, 2011–2023 (2009). https://doi.org/10.1007/s00262-009-0708-5

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