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Activation of CD1d-restricted natural killer T cells can inhibit cancer cell proliferation during chemotherapy by promoting the immune responses in murine mesothelioma

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Abstract

We studied the impact of natural killer T (NKT) cell activation by alpha-galactocysylceramide (α-GalCer, α-GC) on cancer cell repopulation during chemotherapy in murine mesothelioma. The number of NKT cells was found to be increased during the development of murine mesothelioma. NKT cells specifically recognize α-GC through CD1d resulting in their activation and expansion. Tumor-bearing mice were treated with chemotherapy once weekly, and α-GC was followed after each cycle of chemotherapy. Anti-tumor effect was evaluated on wild-type (WT) and CD1d knockout (CD1dKO) mice. Cancer cell proliferation and apoptosis were evaluated by Ki67 and TUNEL immunohistochemistry. CD4+ and CD8+ T cell proportion and activation in tumor, spleen, draining lymph node and peripheral blood were determined by flow cytometry, and gene expression of activated T cell-related cytokines was quantified by reverse transcription PCR. NKT cells were identified by CD1d-α-GC-tetramer staining. In WT mice, tumor growth delay was achieved by cisplatin (Cis), and this effect was improved in combination with α-GC, but α-GC alone had little effect. Cancer cell proliferation during chemotherapy was significantly inhibited by α-GC, while cancer cell death was significantly upregulated. α-GC following chemotherapy resulted in NKT cell expansion and an increase of interferon-γ production in the draining lymph node, blood and spleen. Gene expression of immune-associated cytokines was upregulated. Strikingly, the percentage of inducible T cell co-stimulator+CD4 T cells, Th17/Tc17 cells increased in splenocytes. In CD1d KO mice, however, Cis alone was less effective and Cis + α-GC provided no additional benefit over Cis alone. α-GC alone had minimal effect in both mice. NKT activation between cycles of chemotherapy could improve the outcome of mesothelioma treatment.

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Abbreviations

α-GC:

α-GalCer, alpha-galactocysylceramide

APC:

Antigen-presenting cell

Cis:

Cisplatin

DC:

Dendritic cell

ICOS:

Inducible T cell co-stimulator

IFN:

Interferon

KO:

Knockout

MPM:

Malignant pleural mesothelioma

NKT:

Natural killer T cell

sc:

Subcutaneously

WT:

Wild type

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Acknowledgments

This work was partly supported by the Mesothelioma Foundation at Princess Margaret Hospital, Canada, and the research grant from the Mesothelioma Applied Research Foundation, USA. Dr. Marc de Perrot is the recipient of the grants and the Head of Toronto Mesothelioma Research Program, Canada. We would like to thank the Flow Cytometry Facility of the Hospital for Sick Children and Animal Resources Centre, University Health Network, Toronto, Canada.

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No potential conflict of interests.

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Authors and Affiliations

  1. Latner Thoracic Surgery Research Laboratories, Division of Thoracic Surgery, Toronto General Hospital, University Health Network, University of Toronto, Toronto, ON, Canada

    Licun Wu, Zhihong Yun, Tetsuzo Tagawa, Luis De la Maza, Matthew Onn Wu, Julie Yu, Yidan Zhao & Marc de Perrot

  2. Toronto Mesothelioma Research Program, Division of Thoracic Surgery, Toronto General Hospital, 9N-961, 200 Elizabeth St, Toronto, ON, M5G 2C4, Canada

    Marc de Perrot

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  1. Licun Wu
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Correspondence to Marc de Perrot.

Additional information

Some preliminary results were presented at the 15th World Conference on Lung Cancer (WCLC2013), Sydney, Australia (abstract published in the Journal of Thoracic Oncology, Volume 8, Supplement 2, November 2013).

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Wu, L., Yun, Z., Tagawa, T. et al. Activation of CD1d-restricted natural killer T cells can inhibit cancer cell proliferation during chemotherapy by promoting the immune responses in murine mesothelioma. Cancer Immunol Immunother 63, 1285–1296 (2014). https://doi.org/10.1007/s00262-014-1597-9

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