Neutrophil Microvesicles from Healthy Control and Rheumatoid Arthritis Patients Prevent the Inflammatory Activation of Macrophages

Hefin I Rhys,Adrian Moore,Costantino Pitzalis,Mauro Perretti,Lucy V Norling,Francesco Dell'Accio

doi:10.1016/j.ebiom.2018.02.003

Hefin I Rhys, Adrian Moore + Show 4 more

Open Access

https://doi.org/10.1016/j.ebiom.2018.02.003

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Abstract

Microvesicles (MVs) are emerging as a novel means to enact cell-to-cell communication in inflammation. Here, we aimed to ascertain the ability of neutrophil-derived MVs to modulate target cell behaviour, the focus being the macrophage.MVs were generated in response to tumour necrosis factor-α, from healthy control neutrophils or those from rheumatoid arthritis patients. MVs were used to stimulate human monocyte-derived macrophages in vitro, or administered intra-articularly in the K/BxN mouse model of arthritis. A macrophage/fibroblast-like synoviocyte co-culture system was used to study the effects of vesicles on the crosstalk between these cells.We demonstrate a direct role for phosphatidylserine and annexin-A1 exposed by the MVs to counteract classical activation of the macrophages, and promote the release of transforming growth factor-β, respectively. Classically-activated macrophages exposed to neutrophil MVs no longer activated fibroblast-like synoviocytes in subsequent co-culture settings. Finally, intra-articular administration of neutrophil MVs from rheumatoid arthritis patients in arthritic mice affected the phenotype of joint macrophages.Altogether these data, with the identification of specific MV determinants, open new opportunities to modulate on-going inflammation in the synovia – mainly by affecting macrophage polarization and potentially also fibroblast-like synoviocytes - through the delivery of autologous or heterologous MVs produced from neutrophils.

Highlights

Released directly from the plasma membrane of virtually all cells in response to calcium signalling, microvesicles (MV; termed microparticles or more generically extracellular vesicles) are right-side out, double membrane-enclosed structures with a 100–1000 nm diameter
Neutrophils from 4 donors stimulated with TNF-α produced 7.9 ± 1.6 × 107 vesicles/mL, compared with 2.2 ± 0.5 × 107 quantified in unstimulated cells (Fig. 1D)
ImageStream analysis of 4 distinct preparations showed that 16% ± 7% of TNF-αstimulated MVs expressed phosphatidylserine; 20% ± 6% MVs were annexin A1 (anxA1) positive; 90% ± 4% were stained for CD66b while 0.12% ± 0.04% were CD14 positive, confirming, in essence, the lack of contaminating monocyte MVs

Summary

Introduction

Released directly from the plasma membrane of virtually all cells in response to calcium signalling, microvesicles (MV; termed microparticles or more generically extracellular vesicles) are right-side out, double membrane-enclosed structures with a 100–1000 nm diameter. Present in a variety of biological fluids, MVs can impart both homeostatic and pathophysiological functions on local and distant tissues. Despite their propensity to drive acute inflammation, neutrophils are one of few populations of cells whose MVs are known to promote tissue protection, and in some cases repair, by affecting function and phenotype of target cells In RA, these pro-inflammatory macrophages drive disease progression and cartilage erosion by i) recruiting other immune cells (Misharin et al, 2014; Vogelpoel et al, 2014), ii) promoting fibroblastlike synoviocyte (FLS) activation (Wilkinson et al, 1993), and iii) undergoing osteoclastogenesis (Fujikawa et al, 1996)

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