Assessment of ASC Oligomerization by Flow Cytometry

Area of Science:

• Immunology
• Cellular Biology

Background:

• Inflammasomes are crucial multiprotein complexes in innate and adaptive immunity.
• Upon activation, inflammasomes nucleate the apoptosis-associated speck-like protein containing a CARD (ASC) into large aggregates known as ASC specks.

Purpose of the Study:

• To present a flow cytometry method for monitoring inflammasome activation in human monocytes.
• To detect intracellular ASC oligomers as a readout for inflammasome activation.

Main Methods:

• Utilizing flow cytometry to detect intracellular ASC oligomers.
• Stimulating human monocytes with extracellular ATP to activate the NLRP3 inflammasome.
• Stimulating human monocytes with Clostridium difficile toxin B to activate the pyrin inflammasome.

Main Results:

• The described method allows for the detection of ASC speck formation.
• This technique can differentiate between NLRP3 and pyrin inflammasome activation.

Conclusions:

• The developed flow cytometry method provides a quantifiable way to assess inflammasome activation.
• This approach can be applied to evaluate inflammasome activation in blood samples from patients with chronic inflammatory diseases.