Article

TREM-1 multimerization is essential for its activation on monocytes and neutrophils

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Abstract

The triggering receptor expressed on myeloid cells-1 (TREM-1) is a receptor expressed on innate immune cells. By promoting the amplification of inflammatory signals that are initially triggered by Toll-like receptors (TLRs), TREM-1 has been characterized as a major player in the pathophysiology of acute and chronic inflammatory diseases, such as septic shock, myocardial infarction, atherosclerosis, and inflammatory bowel diseases. However, the molecular events leading to the activation of TREM-1 in innate immune cells remain unknown. Here, we show that TREM-1 is activated by multimerization and that the levels of intracellular Ca2+ release, reactive oxygen species, and cytokine production correlate with the degree of TREM-1 aggregation. TREM-1 activation on primary human monocytes by LPS required a two-step process consisting of upregulation followed by clustering of TREM-1 at the cell surface, in contrast to primary human neutrophils, where LPS induced a rapid cell membrane reorganization of TREM-1, which confirmed that TREM-1 is regulated differently in primary human neutrophils and monocytes. In addition, we show that the ectodomain of TREM-1 is able to homooligomerize in a concentration-dependent manner, which suggests that the clustering of TREM-1 on the membrane promotes its oligomerization. We further show that the adapter protein DAP12 stabilizes TREM-1 surface expression and multimerization. TREM-1 multimerization at the cell surface is also mediated by its endogenous ligand, a conclusion supported by the ability of the TREM-1 inhibitor LR12 to limit TREM-1 multimerization. These results provide evidence for ligand-induced, receptor-mediated dimerization of TREM-1. Collectively, our findings uncover the mechanisms necessary for TREM-1 activation in monocytes and neutrophils.

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Acknowledgements

We thank Sebastien Hupont and Dominique Dumas from the Cellular imaging plateform (PTIBC) at Lorraine University for providing us full access to confocal microscopy and for their technical support. We also thank Jean-Marc Alberto from Inserm U954 for his help with the in situ proximity ligation assay experiments and analysis with the Blobfinder software. We thank Dr. Hortense Mazon for electrospray ionization-mass spectrometry spectra carried out in the Mass Spectometry platform of Université de Lorraine (SCMS). Microcalorimetry and surface plasmon resonance were performed at the Molecular Interactions and Biophysics platform of Université de Lorraine (SCBIM). G.W. and A.J.R.H. are supported by the Roadmap Initiative Proteins@Work (project number 184.032.201), funded by the Netherlands Organization for Scientific Research (NWO).

Author information

Affiliations

  1. INOTREM, Vandœuvre-les-Nancy, France

    • Kevin Carrasco
    • , Amir Boufenzer
    • , Lucie Jolly
    • , Emilie Vinolo
    •  & Marc Derive
  2. UMR-S 1116, Defaillance cardiovasculaire aigue et chronique, Vandœuvre-les-Nancy, France

    • Kevin Carrasco
    • , Lucie Jolly
    •  & Sebastien Gibot
  3. UMR7365, Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA), CNRS-Université de Lorraine, Vandœuvre-les-Nancy, France

    • Helene Le Cordier
  4. Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences and Netherlands Proteomics Center, Utrecht University, Utrecht, The Netherlands

    • Guanbo Wang
    •  & Albert JR Heck
  5. Innate Immunity, German Cancer Research Center (DKFZ), Heidelberg, Germany

    • Adelheid Cerwenka
  6. CovalX, Zurich, Switzerland

    • Alexis Nazabal
  7. Service Commun de Biophysique Interactions Moléculaires (SCBIM), FR3209, Biopôle de l’Université de Lorraine, Vandœuvre-les-Nancy, France

    • Alexandre Kriznik
  8. Inatherys, Evry Cedex, France

    • Pierre Launay

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Competing interests

M.D. and S.G. are co-founders of Inotrem SA, a Company developing TREM-1 inhibitors. P.L. is co-founder of Inatherys SA, a Company developing therapeutic monoclonal antibodies. A.N. is co-founder of CovalX GmbH, a Company providing services for the analysis of protein complexes. The remaining authors declare that they have no competing interests.

Corresponding author

Correspondence to Marc Derive.

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