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Immunization with Antigen-Pulsed Dendritic Cells Against Highly Virulent Cryptococcus gattii Infection: Analysis of Cytokine-Producing T Cells

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Vaccines for Invasive Fungal Infections

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1625))

Abstract

Cryptococcosis caused by highly virulent Cryptococcus gattii (Hv-Cg) is an emerging infectious disease that affects immunocompetent individuals. The Hv-Cg outbreak began in 1999, but the mechanisms responsible for its hyper-virulence as well as protective immunity against Hv-Cg infection remain to be elucidated. To better understand the protective immunity against Hv-Cg infection, we developed a novel immunization method using antigen-pulsed dendritic cells (DCs). We constructed a capsule-deficient Cg strain (∆cap60) and used it as a vaccine antigen. Mouse bone marrow-derived DCs were pulsed with ∆cap60 and transferred into mice twice before pulmonary infection with Hv-Cg strain R265. This DC-based immunization strongly induced cell-mediated immunity, including Th1 cells, Th17 cells, and multinucleated giant cells enclosing fungal cells in lungs. This vaccination significantly ameliorated the fungal burden and the survival rate after pulmonary infection with R265. The efficacy of DC-based immunization was significantly but partially reduced in IFNγ-deficient mice, thereby suggesting that the Th1 and Th17 responses play roles in vaccine-induced protection against Hv-Cg infection. This approach might provide new insights into overcoming Hv-Cg infections in immunocompetent subjects. In this chapter, we describe the procedures for DC-vaccine preparation and the analysis of cytokine-producing CD4+ T cells.

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Acknowledgments

This chapter describes research supported by KAKENHI (15K21644, 16H05349, and 15K15383) from the Ministry of Education, Culture, Sports, Science and Technology of Japan, by the Research program on Emerging and Re-emerging Infectious Diseases from the Japan Agency for Medical Research and Development, AMED, and by LEGEND Research Grant Program 2015 from Tomy Digital Biology Co. Ltd.

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Authors and Affiliations

  1. Department of Chemotherapy and Mycoses, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan

    Keigo Ueno, Makoto Urai, Shogo Takatsuka, Masahiro Abe, Yoshitsugu Miyazaki & Yuki Kinjo

Authors
  1. Keigo Ueno
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  2. Makoto Urai
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  3. Shogo Takatsuka
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  4. Masahiro Abe
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  5. Yoshitsugu Miyazaki
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  6. Yuki Kinjo
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Corresponding author

Correspondence to Yuki Kinjo .

Editor information

Editors and Affiliations

  1. Department of Molecular Immunology, Beckman Research Institute of City of Hope, Duarte, California, USA

    Markus Kalkum

  2. Department of Molecular Immunology, Beckman Research Institute of City of Hope, Duarte, California, USA

    Margarita Semis

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Ueno, K., Urai, M., Takatsuka, S., Abe, M., Miyazaki, Y., Kinjo, Y. (2017). Immunization with Antigen-Pulsed Dendritic Cells Against Highly Virulent Cryptococcus gattii Infection: Analysis of Cytokine-Producing T Cells. In: Kalkum, M., Semis, M. (eds) Vaccines for Invasive Fungal Infections. Methods in Molecular Biology, vol 1625. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7104-6_22

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  • DOI: https://doi.org/10.1007/978-1-4939-7104-6_22

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7103-9

  • Online ISBN: 978-1-4939-7104-6

  • eBook Packages: Springer Protocols

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