Francisella tularensis catalase restricts immune function by impairing TRPM2 channel activity

Nicole L. Shakerley, Akshaya Chandrasekaran, Mohamed Trebak, Barbara A. Miller, J. Andrés Melendez

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

As an innate defense mechanism, macrophages produce reactive oxygen species that weaken pathogens and serve as secondary messengers involved in immune function. The Gram-negative bacterium Francisella tularensis utilizes its antioxidant armature to limit the host immune response, but the mechanism behind this suppression is not defined. Here we establish that F. tularensis limits Ca2+ entry in macrophages, thereby limiting actin reorganization and IL-6 production in a redox-dependent fashion. Wild type (live vaccine strain) or catalase-deficient F. tularensis (AkatG) show distinct profiles in their H2O2 scavenging rates, 1 and 0.015 pM/s, respectively. Murine alveolar macrophages infected with AkatG display abnormally high basal intracellular Ca2+ concentration that did not increase further in response to H2O2. Additionally, AkatG-infected macrophages displayed limited Ca2+ influx in response to ionomycin, as a result of ionophore H2O2 sensitivity. Exogenously added H2O2 or H2O2 generated by AkatG likely oxidizes ionomycin and alters its ability to transport Ca2+. Basal increases in cytosolic Ca2+ and insensitivity to H2O2-mediated Ca2+ entry in AkatG-infected cells are reversed by the Ca2+ channel inhibitors 2-aminoethyl diphenylborinate and SKF-96365. 2-Amino-ethyldiphenylborinate butnotSKF-96365 abrogated AkatG-dependent increases in macrophage actin remodeling and IL-6 secretion, suggesting a role for H2O2-mediated Ca2+ entry through the transient receptor potential melastatin 2 (TRPM2) channel in macrophages. Indeed, increases in basal Ca2+, actin polymerization, and IL-6 production are reversed in TRPM2-null macrophages infected with AkatG. Together, our findings provide compelling evidence thatF. tularensis catalase restricts reactive oxygen species to temper macrophage TRPM2-mediated Ca2+ signaling and limit host immune function.

Original languageEnglish (US)
Pages (from-to)3871-3881
Number of pages11
JournalJournal of Biological Chemistry
Volume291
Issue number8
DOIs
StatePublished - Feb 19 2016

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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