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Published on: March 16, 2018
Modulation and Determination of the Status of Inflammasomes in Leishmania-Infected Macrophages
Arunima Biswas1, Anindita Bhattacharjee2, Pijush K Das3
1Department of Zoology, Molecular Cell Biology laboratory, University of Kalyani, Kalyani, India. arunima10@klyuniv.ac.in.
Abstract:
Leishmania, an intra-macrophage kinetoplastid parasite, modulates a vast array of defensive mechanisms of the host macrophages to create a comfortable environment for their survival. When the host encounters intracellular pathogens, a multimeric protein complex called NLRP3 inflammasome gets turned on, leading to caspase-1 activation-mediated maturation of IL-1β from its pro-form. However, Leishmania often manages to neutralize inflammasome activation by manipulating negative regulatory molecules of the host itself. Exhaustion of NLRP3 and pro-IL-1β result from decreased NF-κB activity in infection, which was attributed to increased expression of A20, a negative regulator of NF-κB signalling. Moreover, reactive oxygen species, another key requirement for inflammasome activation, are inhibited by mitochondrial uncoupling protein 2 (UCP2) which is upregulated by Leishmania. Inflammasome activation is a complex event and procedures involved in monitoring inflammasome activation need to be accurate and error-free. In this chapter, we summarize the protocol that includes various experimental procedures required for the determination of the status of inflammasomes in Leishmania-infected macrophages.
Insights
Leishmania parasites evade host defenses by inhibiting NLRP3 inflammasome activation. This study details methods to accurately assess inflammasome status in infected macrophages.
Area of Science:
- Immunology
- Cell Biology
- Parasitology
Background:
- Leishmania, an intracellular parasite, manipulates host macrophages for survival.
- NLRP3 inflammasome activation, involving caspase-1 and IL-1β, is crucial for host defense against pathogens.
- Leishmania actively suppresses inflammasome activation through host regulatory molecules.
Purpose of the Study:
- To summarize experimental protocols for determining inflammasome activation status.
- To provide accurate and error-free methods for monitoring inflammasome activity in Leishmania-infected macrophages.
Main Methods:
- Detailed experimental procedures for assessing inflammasome activation.
- Focus on monitoring caspase-1 activation and IL-1β maturation.
- Analysis of NF-κB signaling, A20 expression, and reactive oxygen species (ROS) levels.
Main Results:
- Leishmania infection leads to decreased NF-κB activity and NLRP3 inflammasome exhaustion.
- Upregulation of A20 by Leishmania inhibits NF-κB signaling.
- Mitochondrial uncoupling protein 2 (UCP2) upregulation by Leishmania inhibits ROS production, further suppressing inflammasome activation.
Conclusions:
- Leishmania employs sophisticated mechanisms involving A20 and UCP2 to evade host inflammasome-mediated immunity.
- Accurate assessment of inflammasome status is critical for understanding host-parasite interactions and developing therapeutic strategies.
