Multiplexed proteomics of autophagy-deficient murine macrophages reveals enhanced antimicrobial immunity via the oxidative stress response

eLife

Junghyun Lim¹  Timurs Maculins²  John R Rohde³  Erik Verschueren⁴  Trent Hinkle⁴  Meena Choi⁵  Shilpa Rao⁶  Yasin Senbabaoglu⁶  Patrick Chang⁷  Cecile Chalouni⁷  Mike Reichelt⁷  Anand Kumar Katakam⁷  Youngsu Kwon⁸  Brent Mckenzie⁸  Ryan C Kunz⁹  Brian K Erickson⁹  Ivan Dikic^1,10  Donald S Kirkpatrick^1,11  Aditya Murthy^1,11  Ting Huang^1,12  Olga Vitek^1,12  Tsung-Heng Tsai^1,13 

[1] Department of Cancer Immunology, Genentech, South San Francisco, United States;Department of Cancer Immunology, Genentech, South San Francisco, United States;Institute of Biochemistry II, Goethe University, Frankfurt am Main, Germany;Department of Microbiology and Immunology, Dalhousie University, Halifax, Canada;Department of Microchemistry, Proteomics and Lipidomics, Genentech, South San Francisco, United States;Department of Microchemistry, Proteomics and Lipidomics, Genentech, South San Francisco, United States;Khoury College of Computer Sciences, Northeastern University, Boston, United States;Department of Oncology Bioinformatics, Genentech, South San Francisco, United States;Department of Pathology, Genentech, South San Francisco, United States;Department of Translational Immunology, Genentech, South San Francisco, United States;IQ Proteomics LLC, Cambridge, United States;Institute of Biochemistry II, Goethe University, Frankfurt am Main, Germany;Department of Infectious Diseases, Genentech, South San Francisco, United States;Interline Therapeutics, South San Francisco, United States;Khoury College of Computer Sciences, Northeastern University, Boston, United States;Khoury College of Computer Sciences, Northeastern University, Boston, United States;Department of Mathematical Sciences, Kent State University, Kent, United States;

关键词: autophagy;    oxidative damage;    macrophages;    shigella flexneri;    mass spectrometry;    proteomics;    Mouse;    Other;   

DOI  :  10.7554/eLife.62320

来源: eLife Sciences Publications, Ltd

【 摘 要 】

Defective autophagy is strongly associated with chronic inflammation. Loss-of-function of the core autophagy gene Atg16l1 increases risk for Crohn’s disease in part by enhancing innate immunity through myeloid cells such as macrophages. However, autophagy is also recognized as a mechanism for clearance of certain intracellular pathogens. These divergent observations prompted a re-evaluation of ATG16L1 in innate antimicrobial immunity. In this study, we found that loss of Atg16l1 in myeloid cells enhanced the killing of virulent Shigella flexneri (S.flexneri), a clinically relevant enteric bacterium that resides within the cytosol by escaping from membrane-bound compartments. Quantitative multiplexed proteomics of murine bone marrow-derived macrophages revealed that ATG16L1 deficiency significantly upregulated proteins involved in the glutathione-mediated antioxidant response to compensate for elevated oxidative stress, which simultaneously promoted S.flexneri killing. Consistent with this, myeloid-specific deletion of Atg16l1 in mice accelerated bacterial clearance in vitro and in vivo. Pharmacological induction of oxidative stress through suppression of cysteine import enhanced microbial clearance by macrophages. Conversely, antioxidant treatment of macrophages permitted S.flexneri proliferation. These findings demonstrate that control of oxidative stress by ATG16L1 and autophagy regulates antimicrobial immunity against intracellular pathogens.

【 授权许可】

CC BY