The Zebrafish as a New Model for the In Vivo Study of Shigella flexneri Interaction with Phagocytes and Bacterial Autophagy

PLoS Pathogens

Michael Hollinshead¹  Andrea Sirianni²  Jean-Pierre Levraud³  Maria J. Mazon Moya⁴  Pierre Boudinot⁵  Laurent Boucontet⁶  Serge Mostowy⁷  Philippe Herbomel⁷  Pascale Cossart⁷  Emma Colucci-Guyon⁸ 

[1] CNRS, URA2578, Paris, France;INRA, USC2020, Paris, France;INRA, Virologie et Immunologie Moléculaire, Jouy-en-Josas, France;Inserm, U604, Paris, France;Institut Pasteur, Unité Macrophages et Développement de l'Immunité, Département de Biologie du Développement et des Cellules Souches, Paris, France;Institut Pasteur, Unité des Interactions Bactéries-Cellules, Département de Biologie Cellulaire et Infection, Paris, France;Section of Microbiology, MRC Centre for Molecular Bacteriology and Infection, Imperial College London, London, United Kingdom;Section of Virology, Faculty of Medicine, Imperial College London, London, United Kingdom

关键词: Shigellosis;    Larvae;    Zebrafish;    Autophagic cell death;    Macrophages;    Shigella;    Neutrophils;    Septins;   

DOI  :  10.1371/journal.ppat.1003588

学科分类：生物科学（综合）

来源: Public Library of Science

【 摘 要 】

Autophagy, an ancient and highly conserved intracellular degradation process, is viewed as a critical component of innate immunity because of its ability to deliver cytosolic bacteria to the lysosome. However, the role of bacterial autophagy in vivo remains poorly understood. The zebrafish (Danio rerio) has emerged as a vertebrate model for the study of infections because it is optically accessible at the larval stages when the innate immune system is already functional. Here, we have characterized the susceptibility of zebrafish larvae to Shigella flexneri, a paradigm for bacterial autophagy, and have used this model to study Shigella-phagocyte interactions in vivo. Depending on the dose, S. flexneri injected in zebrafish larvae were either cleared in a few days or resulted in a progressive and ultimately fatal infection. Using high resolution live imaging, we found that S. flexneri were rapidly engulfed by macrophages and neutrophils; moreover we discovered a scavenger role for neutrophils in eliminating infected dead macrophages and non-immune cell types that failed to control Shigella infection. We observed that intracellular S. flexneri could escape to the cytosol, induce septin caging and be targeted to autophagy in vivo. Depletion of p62 (sequestosome 1 or SQSTM1), an adaptor protein critical for bacterial autophagy in vitro, significantly increased bacterial burden and host susceptibility to infection. These results show the zebrafish larva as a new model for the study of S. flexneri interaction with phagocytes, and the manipulation of autophagy for anti-bacterial therapy in vivo.

【 授权许可】

CC BY