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Optimized magnitude of cryosurgery facilitating anti-tumor immunoreaction in a mouse model of Lewis lung cancer

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Abstract

Background

Cryosurgery has reemerged as a less invasive local treatment with possible immune-regulatory effects. However, the optimal magnitude of cryosurgery for achieving immune-regulatory responses at abscopal tumor sites remains unclear. We aimed to investigate appropriate magnitude of cryosurgery for this goal using a mouse model.

Methods

C57BL/6J mice were inoculated with Lewis lung carcinoma cells or B16 melanoma cells in bilateral flanks. The left-sided tumor was cryoablated with repeated freeze/thaw cycles either once, twice, or thrice. The peritumoral injections of LPS were performed. Abscopal tumor volumes were measured, immunohistochemistry was performed for CD4, CD8, Foxp3, and Ki-67, and proinflammatory cytokines were measured in lavage fluid of cryoablated tumor.

Results

The growth rate of the abscopal tumor was slowest in the Cryosurgery ×2 group among the five experimental groups. The proportions of CD4+ T cells and CD8+ T cells in the abscopal tumor were also significantly higher in the Cryosurgery ×2 group. The levels of IL-1β, IL-2, IL-6, IL-12β, IFN-γ, and TNF-α in the peritumoral lavage fluid in Cryosurgery ×2 + LPS group were significantly increased compared with the other groups.

Conclusions

This study suggested that achievement of approximately 73 % damaged area in the cryoablated tumor by two cycles of cryosurgery generates the most favorable immune-regulatory response for abscopal tumors via activation of anti-tumor immune cells as well as increased secretion of proinflammatory cytokines.

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Abbreviations

BCG-CWS:

Bacillus Calmette–Guerin cell wall skeleton

CTLA-4:

Cytotoxic T lymphocyte-associated antigen 4

DC:

Dendritic cell

ELISA:

Enzyme-linked immunosorbent assay

LL2:

Lewis lung carcinoma

LPS:

Lipopolysaccharide

NVCA:

Non-viable cancer area

PD-1:

Programmed cell death-1

PD-L1:

Programmed cell death-ligand 1

TIL:

Tumor-infiltrating lymphocyte

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Acknowledgments

This work was supported in part by a Grant-in-Aid for Scientific Research from the Astra Zeneca Science foundation (#201301102).

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Authors and Affiliations

  1. Department of General Thoracic Surgery, Teikyo University School of Medicine, 2-11-1 Kaga, Itabashi-ku, Tokyo, 173-8606, Japan

    Yusuke Takahashi, Noriyuki Matsutani, Hitoshi Dejima, Takashi Nakayama, Ryo Okamura, Hirofumi Uehara & Masafumi Kawamura

  2. Department of General Thoracic Surgery, Saitama Medical Center, Saitama Medical University, Kawagoe, Saitama, Japan

    Yotaro Izumi

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  1. Yusuke Takahashi
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  2. Yotaro Izumi
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  3. Noriyuki Matsutani
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Correspondence to Yusuke Takahashi.

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Takahashi, Y., Izumi, Y., Matsutani, N. et al. Optimized magnitude of cryosurgery facilitating anti-tumor immunoreaction in a mouse model of Lewis lung cancer. Cancer Immunol Immunother 65, 973–982 (2016). https://doi.org/10.1007/s00262-016-1858-x

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  • DOI: https://doi.org/10.1007/s00262-016-1858-x

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