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Cytofluorometric Quantification of Cell Death Elicited by NLR Proteins
Valentina Sica1,2,3,4,5, Gwenola Manic6, Guido Kroemer2,3,7,8,9,5
1Gustave Roussy Cancer Campus, Villejuif, France.
Abstract:
Nucleotide-binding domain and leucine-rich repeat containing (NLR) proteins, also known as NOD-like receptors, are critical components of the molecular machinery that senses intracellular danger signals to initiate an innate immune response against invading pathogens or endogenous sources of hazard. The best characterized effect of NLR signaling is the secretion of various cytokines with immunostimulatory effects, including interleukin (IL)-1β and IL-18. Moreover, at least under specific circumstances, NLRs can promote regulated variants of cell death. Here, we detail two protocols for the cytofluorometric quantification of cell death-associated parameters that can be conveniently employed to assess the lethal activity of specific NLRs or their ligands.
Insights
Nucleotide-binding domain and leucine-rich repeat containing (NLR) proteins are key to innate immunity. This study presents methods to quantify cell death, assessing NLRs
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- Nucleotide-binding domain and leucine-rich repeat containing (NLR) proteins, also known as NOD-like receptors, are crucial for innate immunity.
- NLRs detect intracellular danger signals, initiating immune responses against pathogens and hazardous endogenous materials.
- NLR signaling primarily leads to the release of immunostimulatory cytokines like IL-1β and IL-18.
Purpose of the Study:
- To present two protocols for quantifying cell death parameters using cytofluorometry.
- To provide a method for assessing the cell-killing activity of specific NLRs or their ligands.
Main Methods:
- Cytofluorometric quantification of cell death-associated parameters.
- Detailed protocols for assessing NLR-mediated cell death.
Main Results:
- Established protocols for cytofluorometric assessment of cell death.
- Demonstrated the utility of these methods for evaluating NLR activity.
Conclusions:
- The developed protocols offer a reliable way to measure NLR-induced cell death.
- These methods can be applied to study the function of various NLRs and their impact on cell viability.
