Toxoplasma gondii Chitinase Induces Macrophage Activation

PLoS One. 2015 Dec 10;10(12):e0144507. doi: 10.1371/journal.pone.0144507. eCollection 2015.

Abstract

Toxoplasma gondii is an obligate intracellular protozoan parasite found worldwide that is able to chronically infect almost all vertebrate species, especially birds and mammalians. Chitinases are essential to various biological processes, and some pathogens rely on chitinases for successful parasitization. Here, we purified and characterized a chitinase from T. gondii. The enzyme, provisionally named Tg_chitinase, has a molecular mass of 13.7 kDa and exhibits a Km of 0.34 mM and a Vmax of 2.64. The optimal environmental conditions for enzymatic function were at pH 4.0 and 50 °C. Tg_chitinase was immunolocalized in the cytoplasm of highly virulent T. gondii RH strain tachyzoites, mainly at the apical extremity. Tg_chitinase induced macrophage activation as manifested by the production of high levels of pro-inflammatory cytokines, a pathogenic hallmark of T. gondii infection. In conclusion, to our knowledge, we describe for the first time a chitinase of T. gondii tachyzoites and provide evidence that this enzyme might influence the pathogenesis of T. gondii infection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Chitinases / genetics
  • Chitinases / immunology*
  • Chitinases / metabolism
  • Chromatography, Liquid
  • Cytokines / immunology
  • Cytokines / metabolism
  • Cytoplasm / enzymology
  • Host-Parasite Interactions / immunology
  • Hydrogen-Ion Concentration
  • Inflammation Mediators / immunology
  • Inflammation Mediators / metabolism
  • Kinetics
  • Macrophage Activation / immunology*
  • Macrophages, Peritoneal / immunology*
  • Macrophages, Peritoneal / metabolism
  • Macrophages, Peritoneal / parasitology
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Microscopy, Confocal
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Structure, Tertiary
  • Protozoan Proteins / chemistry
  • Protozoan Proteins / immunology*
  • Tandem Mass Spectrometry
  • Temperature
  • Toxoplasma / enzymology
  • Toxoplasma / immunology*
  • Toxoplasma / physiology

Substances

  • Cytokines
  • Inflammation Mediators
  • Protozoan Proteins
  • Chitinases

Grants and funding

This study was supported by the Fundação de Amparo à Pesquisa do Estado de São Paulo (Grant number 2013/10741-8). Additional financial help was provided by Conselho Nacional de Desenvolvimento Científico e Tecnológico, and Fundação de Apoio ao Ensino, Pesquisa e Assistência do Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto.