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SLAM is a microbial sensor that regulates bacterial phagosome functions in macrophages

Nature Immunology volume 11, pages 920–927 (2010)Cite this article

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Abstract

Phagocytosis is a pivotal process by which macrophages eliminate microorganisms after recognition by pathogen sensors. Here we unexpectedly found that the self ligand and cell surface receptor SLAM functioned not only as a costimulatory molecule but also as a microbial sensor that controlled the killing of Gram-negative bacteria by macrophages. SLAM regulated activity of the NADPH oxidase NOX2 complex and phagolysosomal maturation after entering the phagosome, following interaction with the bacterial outer membrane proteins OmpC and OmpF. SLAM recruited a complex containing the intracellular class III phosphatidylinositol kinase Vps34, its regulatory protein kinase Vps15 and the autophagy-associated molecule beclin-1 to the phagosome, which was responsible for inducing the accumulation of phosphatidylinositol-3-phosphate, a regulator of both NOX2 function and phagosomal or endosomal fusion. Thus, SLAM connects the Gram-negative bacterial phagosome to ubiquitous cellular machinery responsible for the control of bacterial killing.

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Figure 1: SLAM controls in vivo and in vitro killing of Gram-negative bacteria by mouse macrophages.
Figure 2: Defective NOX2 activity in primary macrophages derived from SLAM-deficient mice.
Figure 3: Impaired phagolysosomal maturation in Slamf1−/− macrophages.
Figure 4: Delay in early phagosomal maturation in Slamf1−/− macrophages.
Figure 5: SLAM enters the E. coli–containing phagosome.
Figure 6: SLAM recognizes E. coli and S. typhimurium Sseb− but not S. aureus.
Figure 7: PtdIns(3)P production in phagosomes of primary macrophages is controlled by SLAM.
Figure 8: SLAM recruits the intracellular Vps34–Vps15–beclin-1 complex to the phagosome.

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Acknowledgements

We thank H. Remold, P. Klebba and members of the Terhorst lab for critical review of the manuscript; S. Laroux for help with initial oxidative burst experiments; A. Nohturft, M. Yaffe, P. Klebba and G. Pier for discussions; B. McCormick (University of Massachusetts) for E. coli F18; C. Nagler (University of Chicago) and P. Klemm (Technical University of Denmark) for eGFP-expressing bacteria; H. Nikaido (University of California Berkeley), P. Klebba (University of Oklahoma) and G. Pier (Harvard Medical School) for mutant strains of E. coli and S. aureus; M. Yaffe (Massachusetts Institute of Technology) for the p40-eGFP construct; L. Cantley (Harvard Medical School) for Vps34-Myc; R. Tsien (University of California San Diego) for the mCherry construct; J. Backer (Albert Einstein College of Medicine) for the Vps34-Vps15-V5 construct; P. Hawkins (Babraham Institute) for p40phox-deficient mice; and S. Targan (Cedars-Sinai) for the highly purified OmpC preparation. Supported by the US National Institutes of Health (AI-15066 to C.T., and DK-068181 and DK-003506-20 to H.C.R.) and the Crohn's and Colitis Foundation of America (S.B. and X.R.).

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Author notes

  1. Scott B Berger and Xavier Romero: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Immunology, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA

    Scott B Berger, Xavier Romero, Chunyan Ma, Guoxing Wang, William A Faubion, Gongxian Liao, Ewoud Compeer, Marton Keszei, Ninghai Wang, Jose R Regueiro & Cox Terhorst

  2. Department of Internal Medicine, Mayo Clinic, Rochester, Minnesota, USA

    William A Faubion

  3. Boston Biomedical Research Institute, Watertown, Massachusetts, USA

    Lucia Rameh

  4. Department of Dermatology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA

    Marianne Boes

  5. Inmunologia, Facultad de Medicina, Universidad Complutense, Madrid, Spain

    Jose R Regueiro

  6. Department of Medicine, Gastrointestinal Unit and Center for the Study of Inflammatory Bowel Disease, Massachusetts General Hospital, Harvard Medical School, Massachusetts, USA

    Hans-Christian Reinecker

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Contributions

S.B.B., X.R. and C.T. designed the experiments, analyzed the data and wrote the manuscript; S.B.B. and X.R. did all of the experiments except for the in vivo killing experiments and some of the Jurkat recognition and immunoprecipitation experiments; C.M. did many of the immunoprecipitation experiments; G.W. helped with the confocal microscopy and Jurkat recognition experiments; W.A.F. initiated the killing experiments, was involved in the oxidative burst experiments and set up the Jurkat recognition experiments; G.L. set up the immunoprecipitation; E.C. did some of the Jurkat recognition experiments; M.K. did the restriction fragment length polymorphism analysis and helped make some of the stable transfectants; L.R. did the HPLC analysis; N.W. produced the Slamf1−/− mice; M.B. provided the MHC class II–eGFP mice and helped with analysis of the Slamf1−/− MHC class II–eGFP experiments; J.R.R. helped with production of the SLAM-mCherry construct; H.C.R. helped with the design, data acquisition and interpretation of the confocal microscopy; and C.T. supervised the study.

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Correspondence to Scott B Berger or Cox Terhorst.

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Berger, S., Romero, X., Ma, C. et al. SLAM is a microbial sensor that regulates bacterial phagosome functions in macrophages. Nat Immunol 11, 920–927 (2010). https://doi.org/10.1038/ni.1931

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