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Evaluation of antitumor immunity by a combination treatment of high-dose irradiation, anti-PDL1, and anti-angiogenic therapy in murine lung tumors

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Abstract

C57BL/6 mice implanted in the flank with murine Lewis lung carcinoma cells were randomized into control, anti-angiogenic, anti-PD-L1, radiotherapy (RT), RT + anti-angiogenic, RT + anti-PD-L1, and RT + anti-PD-L1 + anti-angiogenic therapy groups. Immune response and immunophenotyping were determined by flow cytometry. Vasculature analysis after RT and anti-angiogenic therapy was assessed by quantified power Doppler sonography. Antitumor response, survival, and rechallenged tumor growth were evaluated. RT increased PD-L1 expression on CD8+ T, CD4+ T, dendritic, myeloid-derived suppressor cells (MDSCs), and tumor cells and increased PD-1 expression on CD8+ and CD4+ T cells. Anti-angiogenic therapy insignificantly decreased the RT-induced PD-1 expression on CD8+ and CD4+ T cells, implying a weak reversal of the immune-suppressive environment. Transient vessel collapse was observed within days after RT, and blood flow recovered at 1 week after RT. RT + anti-PD-L1 suppressed the tumor growth, improved survival, and prolonged immune memory capable of protecting against tumor recurrence, evidenced by local accumulation of CD8+ T cells and reduction in MDSCs in microenvironment. Similar and more prominent effects were observed when anti-VEGF was added to RT + anti-PDL1 therapies, implying an additive, rather than synergistic, antitumor immunity. Phenotypic analyses revealed that anti-cancer treatments increased the proportion of effector memory T cells in TILs and splenocytes, and RT, alone or in combination with other treatments, further increased the proportion of central memory T cells in splenocytes. These results provide evidence on operating the immunosuppressive tumor environment and offer insights into the design of the new combination treatment.

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Acknowledgements

This study was presented in part at the Radiological Society of North America 104th Scientific Assembly and Annual Meeting (Chicago, USA, November 2019). We thank the staff of the Core Labs, Department of Medical Research, National Taiwan University Hospital, for their technical support.

Funding

This work was supported by the National Taiwan University Hospital (Grant numbers NTUH 107-N4008, 108-N4353, and 109-N4547) and the Ministry of Science and Technology (MST, Taiwan, Contract No. MST 106-2314-B-002-052-MY2).

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Authors and Affiliations

  1. Department of Radiology, National Taiwan University College of Medicine, Taipei, Taiwan

    Jenny Ling-Yu Chen, Yu-Sen Huang & Chun-Wei Wang

  2. Division of Radiation Oncology, Department of Oncology, National Taiwan University Hospital, Taipei, Taiwan

    Jenny Ling-Yu Chen, Chun-Wei Wang & Sung-Hsin Kuo

  3. National Taiwan University Cancer Center, National Taiwan University College of Medicine, Taipei, Taiwan

    Jenny Ling-Yu Chen & Sung-Hsin Kuo

  4. Department of Medical Research, National Taiwan University Hospital, No. 7, Chung-Shan S. Road, Taipei, 100, Taiwan

    Chun-Kai Pan, Ching-Yi Tsai & Yu-Li Lin

  5. Department of Medical Imaging, National Taiwan University Hospital, Taipei, Taiwan

    Yu-Sen Huang

  6. Institute of Toxicology, National Taiwan University College of Medicine, Taipei, Taiwan

    Ching-Yi Tsai

  7. Institute of Biomedical Engineering, College of Medicine and College of Engineering, National Taiwan University, Taipei, Taiwan

    Ming-Jium Shieh

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  1. Jenny Ling-Yu Chen
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Contributions

JLYC, CKP, and YLL conceived and designed the experiments. JLYC, YSH, CKP, CYT, and YLL conducted the experiments. JLYC, CKP, and CWW interpreted the results of the experiments. JLYC, YSH, CWW, and YLL drafted and edited the manuscript. SHK and MJS supervised the study. All authors have read and approved the final version of the manuscript.

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Correspondence to Yu-Li Lin.

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All in vivo experimental protocols were approved by the Institutional Animal Care and Use Committee (IACUC 20160399). Humane endpoints were determined according to a clinical scoring system based on that outlined by the IACUC of our institution.

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Chen, J.LY., Pan, CK., Huang, YS. et al. Evaluation of antitumor immunity by a combination treatment of high-dose irradiation, anti-PDL1, and anti-angiogenic therapy in murine lung tumors. Cancer Immunol Immunother 70, 391–404 (2021). https://doi.org/10.1007/s00262-020-02690-w

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