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Proc Natl Acad Sci U S A. 2019 Apr 12. pii: 201821487. doi: 10.1073/pnas.1821487116. [Epub ahead of print]

Cytosolic Fe-superoxide dismutase safeguards Trypanosoma cruzi from macrophage-derived superoxide radical.

Author information

1
Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, 11800 Montevideo, Uruguay.
2
Centro de Investigaciones Biomédicas (CEINBIO), Facultad de Medicina, Universidad de la República, 11800 Montevideo, Uruguay.
3
Laboratory of Host-Pathogen Interactions-Unidad de Biología Molecular, Institut Pasteur de Montevideo, 11800 Montevideo, Uruguay.
4
Departamento de Bioquímica, Facultad de Medicina, Universidad de la República, 11800 Montevideo, Uruguay; rradi@fmed.edu.uy lpiacenza@fmed.edu.uy.

Abstract

Trypanosoma cruzi, the causative agent of Chagas disease (CD), contains exclusively Fe-dependent superoxide dismutases (Fe-SODs). During T. cruzi invasion to macrophages, superoxide radical (O2 •-) is produced at the phagosomal compartment toward the internalized parasite via NOX-2 (gp91-phox) activation. In this work, T. cruzi cytosolic Fe-SODB overexpressers (pRIBOTEX-Fe-SODB) exhibited higher resistance to macrophage-dependent killing and enhanced intracellular proliferation compared with wild-type (WT) parasites. The higher infectivity of Fe-SODB overexpressers compared with WT parasites was lost in gp91-phox -/- macrophages, underscoring the role of O2 •- in parasite killing. Herein, we studied the entrance of O2 •- and its protonated form, perhydroxyl radical [(HO2 ); pKa = 4.8], to T. cruzi at the phagosome compartment. At the acidic pH values of the phagosome lumen (pH 5.3 ± 0.1), high steady-state concentrations of O2 •- and HO2 were estimated (∼28 and 8 µM, respectively). Phagosomal acidification was crucial for O2 •- permeation, because inhibition of the macrophage H+-ATPase proton pump significantly decreased O2 •- detection in the internalized parasite. Importantly, O2 •- detection, aconitase inactivation, and peroxynitrite generation were lower in Fe-SODB than in WT parasites exposed to external fluxes of O2 •- or during macrophage infections. Other mechanisms of O2 •- entrance participate at neutral pH values, because the anion channel inhibitor 5-nitro-2-(3-phenylpropylamino) benzoic acid decreased O2 •- detection. Finally, parasitemia and tissue parasite burden in mice were higher in Fe-SODB-overexpressing parasites, supporting the role of the cytosolic O2 •--catabolizing enzyme as a virulence factor for CD.

KEYWORDS:

Trypanosoma cruzi; oxidant; superoxide dismutase; superoxide radical; virulence

PMID:
30979807
DOI:
10.1073/pnas.1821487116

Conflict of interest statement

The authors declare no conflict of interest.

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