An optimized whole blood assay measuring expression and activity of NLRP3-, NLRC4 and AIM2-inflammasomes

L Grinstein,Aab Robertson,S Winkler,A Rösen-Wolff,Ma Cooper,H Luksch

doi:10.1186/1546-0096-13-s1-o51

L Grinstein, Aab Robertson + Show 4 more

Open Access

https://doi.org/10.1186/1546-0096-13-s1-o51

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Abstract

Caspase-1 activates proIL-1β and proIL-18 and plays a fundamental role in innate immunity. This pro-inflammatory immune response is required for the initiation of pathogen clearance. Caspase-1 itself is activated in so-called “inflammasomes” which are assembled in response to distinct pathogen associated molecular patterns (PAMP) or danger associated molecular patterns (DAMP). Most studies analyzing the inflammasome/caspase-1 activity of patients ex vivo use PBMC-based assays. Beside monocytes and macrophages, caspase-1 is also activated in PMNs representing the major leukocyte subset in peripheral blood. Thus, analyzing purified PBMCs seems to be highly artificial by excluding caspase-1 activity in PMNs and ignoring cellular interactions. Furthermore, PBMC purification requires large amounts of blood, thereby rendering the assay impractical for the use in small children or neonates.

Highlights

Caspase-1 activates proIL-1b and proIL-18 and plays a fundamental role in innate immunity
Caspase-1 itself is activated in socalled “inflammasomes” which are assembled in response to distinct pathogen associated molecular patterns (PAMP) or danger associated molecular patterns (DAMP)
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Summary

Introduction

Caspase-1 activates proIL-1b and proIL-18 and plays a fundamental role in innate immunity. This pro-inflammatory immune response is required for the initiation of pathogen clearance. Caspase-1 itself is activated in socalled “inflammasomes” which are assembled in response to distinct pathogen associated molecular patterns (PAMP) or danger associated molecular patterns (DAMP). Most studies analyzing the inflammasome/caspase-1 activity of patients ex vivo use PBMC-based assays. Caspase-1 is activated in PMNs representing the major leukocyte subset in peripheral blood. Analyzing purified PBMCs seems to be highly artificial by excluding caspase-1 activity in PMNs and ignoring cellular interactions. PBMC purification requires large amounts of blood, thereby rendering the assay impractical for the use in small children or neonates

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