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Legionella pp 305-319 | Cite as

Inflammasome Activation in Legionella-Infected Macrophages

  • Danielle P. A. Mascarenhas
  • Dario S. ZamboniEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1921)

Abstract

Legionella pneumophila is a gram-negative bacterium that infects many species of unicellular protozoa in freshwater environments. The human infection is accidental, and the bacteria may not have evolved strategies to bypass innate immune signaling in mammalian macrophages. Thus, L. pneumophila triggers many innate immune pathways including inflammasome activation. The inflammasomes are multimolecular platforms assembled in the host cell cytoplasm and lead to activation of inflammatory caspases. Inflammasome activation leads to secretion of inflammatory cytokines, such as IL-1β and IL-18, and an inflammatory form of cell death called pyroptosis, which initiates with the induction of a pore in the macrophage membranes. In this chapter we provide detailed protocols to evaluate Legionella-induced inflammasome activation in macrophages, including real-time pore formation assay, western blotting to detect activation of inflammatory caspases (cleavage and pulldown), and the measurement of inflammatory cytokines.

Key words

Macrophages Inflammasome Caspase-1 Caspase-11 Legionella 

Notes

Acknowledgments

We thank Dr. Alexandre L.N. Silva and Talita D. Fernandes for details and discussions about the protocols detailed in this manuscript. Work in our laboratory is funded by grants from FAPESP and CNPq. We are grateful to technical assistance of Maira Nakamura, Catarina Horta, Leticia Corsi, Victoria Maria dos Santos, Vitoria Turin, and Laís de Castro Batista.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Danielle P. A. Mascarenhas
    • 1
  • Dario S. Zamboni
    • 1
    Email author
  1. 1.Department of Cell Biology, School of Medicine of Ribeirão PretoUniversity of São PauloRibeirão PretoBrazil

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