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.
Leishmania parasites are the causative agent of leishmaniasis, a wide spread disease in tropical and subtropical areas. The microorganisms have been shown to be well-adapted to their hosts and are able to enter their target cells where they replicate themselves. To ensure these processes, Leishmania disrupts a multitude of cellular signals and protective mechanisms, which overall attenuates immune responses against the parasites. A key factor for inflammatory processes, also during infections, is IL-1β. As previous studies suggested a dysregulation of IL-1β levels after infection with Leishmania parasites, we herein investigated the underlying mechanisms. Our work reveals that Leishmania suppressing IL-1β production through its virulence factor GP63. Furthermore, our data suggests that the parasites can dampen the maturation of IL-1β after different stimuli. In this regard we established a role for the suppression of the kinase PKC and the generation of reactive oxygen species, as well as the cleavage of cellular proteins that are important for IL-1β-generation. Thus, we here present a novel aspect for how Leishmania parasites can counteract host protective mechanisms.