S100A8/A9 Is a Marker for the Release of Neutrophil Extracellular Traps and Induces Neutrophil Activation.

Cathelijn E. M. Aarts,Hans Janssen,Judith Zandstra,Evelien G. G. Sprenkeler,Bibian Verstegen,Ines Goetschalckx,Taco W. Kuijpers,Robin van Bruggen,Anton T. J. Tool,Gerard van Mierlo,Nadine D. van Kleef,Ilse Jongerius

doi:10.3390/cells11020236

Abstract

Neutrophils are the most abundant innate immune cells in the circulation and they are the first cells recruited to sites of infection or inflammation. Almost half of the intracellular protein content in neutrophils consists of S100A8 and S100A9, though there has been controversy about their actual localization. Once released extracellularly, these proteins are thought to act as damage-associated molecular patterns (DAMPs), though their mechanism of action is not well understood. These S100 proteins mainly form heterodimers (S100A8/A9, also known as calprotectin) and this heterocomplex is recognized as a useful biomarker for several inflammatory diseases. We observed that S100A8/A9 is highly present in the cytoplasmic fraction of neutrophils and is not part of the granule content. Furthermore, we found that S100A8/A9 was not released in parallel with granular content but upon the formation of neutrophil extracellular traps (NETs). Accordingly, neutrophils of patients with chronic granulomatous disease, who are deficient in phorbol 12-myristate 13-acetate (PMA)-induced NETosis, did not release S100A8/A9 upon PMA stimulation. Moreover, we purified S100A8/A9 from the cytoplasmic fraction of neutrophils and found that S100A8/A9 could induce neutrophil activation, including adhesion and CD11b upregulation, indicating that this DAMP might amplify neutrophil activation.

Highlights

The most abundant innate immune cells in the circulation consist of neutrophilic granulocytes
While S100A8/A9 lacks signal peptides required for secretion via the classical ER/Golgi route [17], it has been suggested that S100A8/A9 is localized within the specific granules of neutrophils and that stimulation of specific granule exocytosis resulted in the release of S100A8/A9 [19]
S100A8/A9 is recognized as a useful biomarker for several inflammatory diseases and is thought to act as damage-associated molecular patterns (DAMPs) once released extracellularly

Summary

Introduction

The most abundant innate immune cells in the circulation consist of neutrophilic granulocytes. They circulate through the blood and are recruited to tissues during infection or inflammation. They are highly mobile, short-lived cells and comprise the first line of defense in the innate immune system [1]. When neutrophils are recruited to the site of infection, they recognize pathogen-associated molecular patterns (PAMPs) or damage-associated molecular patterns (DAMPs), which activate specific pattern recognition receptors (PRRs) on their surface. While PAMPs are derived from microbial molecules, DAMPs are endogenous molecules released during tissue damage [2]. Neutrophils are capable of eliminating pathogens by several mechanisms, i.e., phagocytosis, production of reactive oxygen species (ROS), degranulation, and the formation of neutrophil extracellular traps (NETs) [3]

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