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Inducing Pyroptosis with FlaTox, RodTox, or NeedleTox.
William P Scott1, Isabella Rauch2
1Department of Molecular Microbiology and Immunology, Oregon Health & Science University, Portland, OR, USA.
Methods in Molecular Biology (Clifton, N.J.)
|April 19, 2023
Summary
This study details methods for activating the NAIP-NLRC4 inflammasome using FlaTox systems. It outlines in vitro and in vivo experimental procedures and key measurements for studying pyroptosis.
Area of Science:
- Immunology
- Cellular Biology
Background:
- The NAIP-NLRC4 inflammasome is crucial for studying pyroptosis.
- Targeted activation is key to understanding its role.
Purpose of the Study:
- To describe methods for stimulating the NAIP-NLRC4 inflammasome in vitro and in vivo.
- To provide guidance on experimental setup and considerations for macrophage treatment and systemic inflammasome activation models.
Main Methods:
- Utilized FlaTox and LFn-NAIP-ligand delivery systems for cytosolic delivery.
- Described in vitro stimulation of macrophages and in vivo injections in a murine model.
- Detailed measurement of propidium iodide uptake, LDH release, hematocrit, and body temperature.
Main Results:
- Established protocols for targeted NAIP-NLRC4 inflammasome activation.
- Demonstrated feasibility of in vitro and in vivo stimulation and readout measurements.
- Provided a framework for interrogating inflammasome ligand recognition and downstream effects.
Conclusions:
- The described methods enable robust investigation of the NAIP-NLRC4 inflammasome pathway.
- These techniques facilitate the study of pyroptosis and related inflammatory responses.
- This work offers a valuable resource for researchers in immunology and cell biology.
