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In Vivo Cytotoxicity by α-GalCer-transactivated NK Cells

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Invariant Natural Killer T-Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2388))

Abstract

Invariant natural killer T (iNKT) cells are innate-like, lipid-reactive T lymphocytes known for their potent immunomodulatory properties. In addition to expressing and utilizing cytolytic effector molecules of their own against certain target cells, iNKT cells can be stimulated with α-galactosylceramide (α-GalCer) to augment the cytotoxic capacity of natural killer (NK) cells. Herein, we describe a flow cytometry-based in vivo killing assay that enables examination of α-GalCer-promoted cytotoxicity against β2 microglobulin knockout (β2M−/−) target cells, which mimic tumor and virus-infected cells displaying little to no MHC class I molecules on their surface. Using an anti-asialo GM1 antibody, which depletes NK cells but not iNKT cells, we confirmed that the increased clearance of β2M−/− cells in α-GalCer-primed recipients was mediated by NK cells. The protocol detailed here can be leveraged to assess the functional fitness of iNKT cells and their crosstalk with NK cells and to further our understanding of α-GalCer-promoted cytotoxicity in preclinical immunotherapeutic applications.

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Acknowledgments

Our ongoing studies on invariant T cells are supported through operating grants provided to SMMH by the Canadian Institutes of Health Research (PJT-156295), the Natural Sciences and Engineering Research Council of Canada (NSERC) (04706-2019 RGPIN), and the Canadian Cancer Society (706396). PTR was an NSERC Alexander Graham Bell Canada Graduate Scholar (CGS—Doctoral).

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Authors and Affiliations

  1. Department of Microbiology and Immunology, Western University, London, ON, Canada

    Patrick T. Rudak & S. M. Mansour Haeryfar

  2. Department of Medicine, Division of Clinical Immunology and Allergy, Western University, London, ON, Canada

    S. M. Mansour Haeryfar

  3. Department of Surgery, Division of General Surgery, Western University, London, ON, Canada

    S. M. Mansour Haeryfar

  4. Centre for Human Immunology, Western University, London, ON, Canada

    S. M. Mansour Haeryfar

  5. Lawson Health Research Institute, London, ON, Canada

    S. M. Mansour Haeryfar

Authors
  1. Patrick T. Rudak
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  2. S. M. Mansour Haeryfar
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Corresponding author

Correspondence to S. M. Mansour Haeryfar .

Editor information

Editors and Affiliations

  1. Department of Pathogen Biology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China

    Chaohong Liu

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Rudak, P.T., Haeryfar, S.M.M. (2021). In Vivo Cytotoxicity by α-GalCer-transactivated NK Cells. In: Liu, C. (eds) Invariant Natural Killer T-Cells. Methods in Molecular Biology, vol 2388. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1775-5_15

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  • DOI: https://doi.org/10.1007/978-1-0716-1775-5_15

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1774-8

  • Online ISBN: 978-1-0716-1775-5

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