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Assessing Phagocytosis of Cryptococcus neoformans Cells in Human Monocytes or the J774 Murine Macrophage Cell Line

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Cryptococcus neoformans

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2775))

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Abstract

Monocyte/macrophage cells play a central role in innate immunity against C. neoformans and C. gattii, species known to cause human disease. Cryptococcus is the only fungal genus known to possess such a large extracellular polysaccharide capsule, which impacts interactions of innate cells with the yeast. This interaction results in different fates, such as phagocytosis and intracellular proliferation and, as the interaction progresses, vomocytosis, cell-to-cell transfer, lysis of macrophages, or yeast killing. Differentiating internalized versus external Cryptococcus cells is thus essential to evaluate monocyte–macrophage phagocytosis. We describe here a protocol that allows quantification of Cryptococcus spp. phagocytosis using quantitative flow cytometry in human monocytes and a murine macrophage cell line (J774).

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Acknowledgments

The authors thank Marion Benazra for her helpful support and Professor Arturo Casadevall for providing the 18B7 mAb.

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Authors and Affiliations

  1. Translational Mycology Research Group, Institut Pasteur, Université Paris Cité, National Reference Center for Invasive Mycoses and Antifungals, Mycology Department, Paris, France

    Emmanuel Lafont, Fanny Lanternier & Alexandre Alanio

  2. MRC Centre for Medical Mycology, College of Health and Medicine, University of Exeter, Exeter, Devon, UK

    Carolina Coelho & Fanny Lanternier

  3. Service de maladies infectieuses et tropicales, hôpital Necker-Enfants Malades, Paris, France

    Fanny Lanternier

  4. Laboratoire de parasitologie-mycologie, AP-HP, Hôpital Saint-Louis, Paris, France

    Alexandre Alanio

  5. Department of Mycology, Translational Mycology Group, Institut Pasteur, Université Paris Cité, National Reference Center for Invasive Mycoses and Antifungals, Paris, France

    Aude Sturny-Leclère

Authors
  1. Emmanuel Lafont
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  2. Aude Sturny-Leclère
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  3. Carolina Coelho
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  4. Fanny Lanternier
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  5. Alexandre Alanio
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Corresponding author

Correspondence to Alexandre Alanio .

Editor information

Editors and Affiliations

  1. College of Osteopathic Medicine, Marian University - Indiana, Indianapolis, IN, USA

    Erin E. McClelland

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Lafont, E., Sturny-Leclère, A., Coelho, C., Lanternier, F., Alanio, A. (2024). Assessing Phagocytosis of Cryptococcus neoformans Cells in Human Monocytes or the J774 Murine Macrophage Cell Line. In: McClelland, E.E. (eds) Cryptococcus neoformans. Methods in Molecular Biology, vol 2775. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3722-7_11

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  • DOI: https://doi.org/10.1007/978-1-0716-3722-7_11

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3721-0

  • Online ISBN: 978-1-0716-3722-7

  • eBook Packages: Springer Protocols

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