title: PKC/ROS-Mediated NLRP3 Inflammasome Activation Is Attenuated by Leishmania Zinc-Metalloprotease during Infection.
abstract: Parasites of the Leishmania genus infect and survive within macrophages by inhibiting several microbicidal molecules, such as nitric oxide and pro-inflammatory cytokines. In this context, various species of Leishmania have been reported to inhibit or reduce the production of IL-1β both in vitro and in vivo. However, the mechanism whereby Leishmania parasites are able to affect IL-1β production and secretion by macrophages is still not fully understood. Dependent on the stimulus at hand, the maturation of IL-1β is facilitated by different inflammasome complexes. The NLRP3 inflammasome has been shown to be of pivotal importance in the detection of danger molecules such as inorganic crystals like asbestos, silica and malarial hemozoin, (HZ) as well as infectious agents. In the present work, we investigated whether Leishmania parasites modulate NLRP3 inflammasome activation. Using PMA-differentiated THP-1 cells, we demonstrate that Leishmania infection effectively inhibits macrophage IL-1β production upon stimulation. In this context, the expression and activity of the metalloprotease GP63 - a critical virulence factor expressed by all infectious Leishmania species - is a prerequisite for a Leishmania-mediated reduction of IL-1β secretion. Accordingly, L. mexicana, purified GP63 and GP63-containing exosomes, caused the inhibition of macrophage IL-1β production. Leishmania-dependent suppression of IL-1β secretion is accompanied by an inhibition of reactive oxygen species (ROS) production that has previously been shown to be associated with NLRP3 inflammasome activation. The observed loss of ROS production was due to an impaired PKC-mediated protein phosphorylation. Furthermore, ROS-independent inflammasome activation was inhibited, possibly due to an observed GP63-dependent cleavage of inflammasome and inflammasome-related proteins. Collectively for the first time, we herein provide evidence that the protozoan parasite Leishmania, through its surface metalloprotease GP63, can significantly inhibit NLRP3 inflammasome function and IL-1β production.
authors.affiliation: Department of Microbiology and Immunology, McGill University, Montréal, Québec, Canada; Department of Microbiology, Immunology and Parasitology, Universidade Federal de São Paulo, São Paulo, Brazil.
authors.forename: Marina Tiemi
authors.initials: MT
authors.lastname: Shio
authors.affiliation: Department of Microbiology and Immunology, McGill University, Montréal, Québec, Canada; McGill International Tuberculosis (TB) Centre and the Research Institute of the McGill University Health Centre, Montréal, Québec, Canada.
authors.forename: Jan Gregor
authors.initials: JG
authors.lastname: Christian
authors.affiliation: Department of Microbiology and Immunology, McGill University, Montréal, Québec, Canada.
authors.forename: Jee Yong
authors.initials: JY
authors.lastname: Jung
authors.affiliation: Chicago Medical School, Rosalind Franklin University of Medicine and Science, North Chicago, Illinois, United States of America.
authors.forename: Kwang-Poo
authors.initials: KP
authors.lastname: Chang
authors.affiliation: Department of Microbiology and Immunology, McGill University, Montréal, Québec, Canada; McGill International Tuberculosis (TB) Centre and the Research Institute of the McGill University Health Centre, Montréal, Québec, Canada.
authors.forename: Martin
authors.initials: M
authors.lastname: Olivier
mesh_terms: D000818:Animals; D002352:Carrier Proteins / Q000276:immunology*; D002460:Cell Line; D006801:Humans; D058847:Inflammasomes / Q000276:immunology* / Q000378:metabolism; D053583:Interleukin-1beta / Q000378:metabolism; D007891:Leishmania / Q000201:enzymology* / Q000276:immunology; D007894:Leishmania mexicana / Q000201:enzymology / Q000276:immunology; D007896:Leishmaniasis / Q000276:immunology* / Q000378:metabolism; D008264:Macrophages / Q000276:immunology; D008666:Metalloendopeptidases / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; D010766:Phosphorylation; D011493:Protein Kinase C / Q000276:immunology / Q000378:metabolism; D015800:Protozoan Proteins / Q000276:immunology / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000276:immunology / Q000378:metabolism; D015032:Zinc / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D058847:Inflammasomes; D053583:Interleukin-1beta; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; C453881:NLRP3 protein, human; D015800:Protozoan Proteins; D017382:Reactive Oxygen Species; D011493:Protein Kinase C; D008666:Metalloendopeptidases; C055146:glycoprotein gp63, Leishmania; D015032:Zinc