Identification of Piecemeal Degranulation and Vesicular Transport of MBP-1 in Liver-Infiltrating Mouse Eosinophils During Acute Experimental Schistosoma mansoni Infection.

Felipe F Dias,Gabriel S C Rodrigues,Rossana C N Melo,Vivian V Costa,Florence M Rosa,Hélio Chiarini-Garcia,Peter F Weller,Kássia K Malta,Thiago P Silva,Kátia B Amaral,Gisele O L Rodrigues

doi:10.3389/fimmu.2018.03019

Abstract

Eosinophils have been long associated with helminthic infections, although their functions in these diseases remain unclear. During schistosomiasis caused by the trematode Schistosoma mansoni, eosinophils are specifically recruited and migrate to sites of granulomatous responses where they degranulate. However, little is known about the mechanisms of eosinophil secretion during this disease. Here, we investigated the degranulation patterns, including the cellular mechanisms of major basic protein-1 (MBP-1) release, from inflammatory eosinophils in a mouse model of S. mansoni infection (acute phase). Fragments of the liver, a major target organ of this disease, were processed for histologic analyses (whole slide imaging), conventional transmission electron microscopy (TEM), and immunonanogold EM using a pre-embedding approach for precise localization of major basic protein 1 (MBP-1), a typical cationic protein stored pre-synthesized in eosinophil secretory (specific) granules. A well-characterized granulomatous inflammatory response with a high number of infiltrating eosinophils surrounding S. mansoni eggs was observed in the livers of infected mice. Moreover, significant elevations in the levels of plasma Th2 cytokines (IL-4, IL-13, and IL-10) and serum enzymes (alanine aminotransferase and aspartate aminotransferase) reflecting altered liver function were detected in response to the infection. TEM quantitative analyses revealed that while 19.1% of eosinophils were intact, most of them showed distinct degranulation processes: cytolysis (13.0%), classical and/or compound exocytosis identified by granule fusions (1.5%), and mainly piecemeal degranulation (PMD) (66.4%), which is mediated by vesicular trafficking. Immunonanogold EM showed a consistent labeling for MBP-1 associated with secretory granules. Most MBP-1-positive granules had PMD features (79.0 ± 4.8%). MBP-1 was also present extracellularly and on vesicles distributed in the cytoplasm and attached to/surrounding the surface of emptying granules. Our data demonstrated that liver-infiltrating mouse eosinophils are able to degranulate through different secretory processes during acute experimental S. mansoni infections with PMD being the predominant mechanism of eosinophil secretion. This means that a selective secretion of MBP-1 is occurring. Moreover, our study demonstrates, for the first time, a vesicular trafficking of MBP-1 within mouse eosinophils elicited by a helminth infection. Vesicle-mediated secretion of MBP-1 may be relevant for the rapid release of small concentrations of MBP-1 under cell activation.

Highlights

Eosinophils are innate immune cells with a broad distribution in tissues and notably associated with allergic and helminth parasitic diseases [reviewed in [1,2,3]]
Our study describes marked eosinophil degranulation in vivo in the liver triggered by schistosomiasis mansoni in mice and identifies, for the first time, PMD as the main mode of eosinophil secretion
We provide direct evidence that MBP1 is transported in the cytoplasm of infiltrating eosinophils and released through a vesicular trafficking in response to the acute infection

Summary

Introduction

Eosinophils are innate immune cells with a broad distribution in tissues and notably associated with allergic and helminth parasitic diseases [reviewed in [1,2,3]]. The etiological agents of human schistosomiasis are trematode worms of the genus Schistosoma with most species, including Schistosoma mansoni, the only one that occurs in the Americas, affecting mainly the liver and the intestines [8]. Human infection with this parasite causes significant chronic morbidity with the development of a granulomatous reaction and severe tissue inflammation, which can lead to life-threatening hepatosplenomegaly [reviewed in [9]]. It remains uncertain if eosinophils act as major effector cells against the parasite; as immunomodulators of the immune response; as participants in tissue homeostasis and metabolism, which could favor establishment and maintenance of parasitic worms in their hosts, or merely as operators in remodeling and clearance of debris following injury [7, 10,11,12,13,14]

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