Endothelial junctional membrane protrusions serve as hotspots for neutrophil transmigration

  1. Janine JG Arts
  2. Eike K Mahlandt
  3. Max Grönloh
  4. Lilian Schimmel
  5. Ivar Noordstra
  6. Emma Gordon
  7. Abraham CI van Steen
  8. Simon Tol
  9. Barbara Walzog
  10. Jos van Rijssel
  11. Martijn A Nolte
  12. Marten Postma
  13. Satya Khuon
  14. John M Heddleston
  15. Eric Wait
  16. Teng Leong Chew
  17. Mark Winter
  18. Eloi Montanez
  19. Joachim Goedhart
  20. Jaap D van Buul  Is a corresponding author
  1. Sanquin Research and Landsteiner Laboratory, Netherlands
  2. SILS/UvA, Netherlands
  3. Uppsala University, Sweden
  4. Ludwig-Maximilians-Universität München, Germany
  5. Janelia Research Campus, United States
  6. Howard Hughes Medical Institute, United States
  7. University of Haifa, Israel
  8. University of Barcelona, Spain

Abstract

Upon inflammation, leukocytes rapidly transmigrate across the endothelium to enter the inflamed tissue. Evidence accumulates that leukocytes use preferred exit sites, though it is not yet clear how these hotspots in the endothelium are defined and how they are recognized by the leukocyte. Using lattice light sheet microscopy, we discovered that leukocytes prefer endothelial membrane protrusions at cell junctions for transmigration. Phenotypically, these junctional membrane protrusions are present in an asymmetric manner, meaning that one endothelial cell shows the protrusion and the adjacent one does not. Consequently, leukocytes cross the junction by migrating underneath the protruding endothelial cell. These protrusions depend on Rac1 activity and by using a photo-activatable Rac1 probe, we could artificially generate local exit-sites for leukocytes. Overall, we have discovered a new mechanism that uses local induced junctional membrane protrusions to facilitate/steer the leukocyte escape/exit from inflamed vessel walls.

Data availability

All data generated or analyzed during this study are included in the manuscript and supporting files. Source data files will be provided for Figures 4 and 6.

Article and author information

Author details

  1. Janine JG Arts

    Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  2. Eike K Mahlandt

    Molecular Cytology, SILS/UvA, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  3. Max Grönloh

    Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0109-8225
  4. Lilian Schimmel

    Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0569-0464
  5. Ivar Noordstra

    Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  6. Emma Gordon

    Immunology, Genetics and Pathology, Uppsala University, Uppsala, Sweden
    Competing interests
    The authors declare that no competing interests exist.
  7. Abraham CI van Steen

    Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  8. Simon Tol

    Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  9. Barbara Walzog

    Ludwig-Maximilians-Universität München, Planegg- Martinsried, Germany
    Competing interests
    The authors declare that no competing interests exist.
  10. Jos van Rijssel

    Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8077-1371
  11. Martijn A Nolte

    Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  12. Marten Postma

    Molecular Cytology, SILS/UvA, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
  13. Satya Khuon

    Advanced Imaging Center, Janelia Research Campus, Ashburn, United States
    Competing interests
    The authors declare that no competing interests exist.
  14. John M Heddleston

    Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States
    Competing interests
    The authors declare that no competing interests exist.
  15. Eric Wait

    Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States
    Competing interests
    The authors declare that no competing interests exist.
  16. Teng Leong Chew

    Janelia Research Campus, Howard Hughes Medical Institute, Ashburn, United States
    Competing interests
    The authors declare that no competing interests exist.
  17. Mark Winter

    Department of Marine Sciences, University of Haifa, Haifa, Israel
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1180-1957
  18. Eloi Montanez

    Department of Physiological Sciences, University of Barcelona, Barcelona, Spain
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4059-5056
  19. Joachim Goedhart

    Molecular Cytology, SILS/UvA, Amsterdam, Netherlands
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-0630-3825
  20. Jaap D van Buul

    Molecular Cell Biology, Sanquin Research and Landsteiner Laboratory, Amsterdam, Netherlands
    For correspondence
    j.vanbuul@sanquin.nl
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-0054-7949

Funding

LSBR (1649)

  • Abraham CI van Steen

NWO-ZonMW Vici (91819632)

  • Max Grönloh
  • Jaap D van Buul

Spanish Ministry of Science, Innovation and Universities (PID2019-108902GB-I00)

  • Eloi Montanez

NWO ALW-OPEN (ALWOP.306)

  • Eike K Mahlandt

Deutsche Forschungsgemeinschaft (SFB 914/A02)

  • Barbara Walzog

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All animal experiments were conducted in accordance with German federal animal protection laws and were approved by the Bavarian Government (Regierung von Oberbayern, Munich, Germany)

Copyright

© 2021, Arts et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 3,131
    views
  • 434
    downloads
  • 33
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Janine JG Arts
  2. Eike K Mahlandt
  3. Max Grönloh
  4. Lilian Schimmel
  5. Ivar Noordstra
  6. Emma Gordon
  7. Abraham CI van Steen
  8. Simon Tol
  9. Barbara Walzog
  10. Jos van Rijssel
  11. Martijn A Nolte
  12. Marten Postma
  13. Satya Khuon
  14. John M Heddleston
  15. Eric Wait
  16. Teng Leong Chew
  17. Mark Winter
  18. Eloi Montanez
  19. Joachim Goedhart
  20. Jaap D van Buul
(2021)
Endothelial junctional membrane protrusions serve as hotspots for neutrophil transmigration
eLife 10:e66074.
https://doi.org/10.7554/eLife.66074

Share this article

https://doi.org/10.7554/eLife.66074

Further reading

    1. Cell Biology
    2. Immunology and Inflammation
    Alejandro Rosell, Agata Adelajda Krygowska ... Esther Castellano Sanchez
    Research Article

    Macrophages are crucial in the body’s inflammatory response, with tightly regulated functions for optimal immune system performance. Our study reveals that the RAS–p110α signalling pathway, known for its involvement in various biological processes and tumourigenesis, regulates two vital aspects of the inflammatory response in macrophages: the initial monocyte movement and later-stage lysosomal function. Disrupting this pathway, either in a mouse model or through drug intervention, hampers the inflammatory response, leading to delayed resolution and the development of more severe acute inflammatory reactions in live models. This discovery uncovers a previously unknown role of the p110α isoform in immune regulation within macrophages, offering insight into the complex mechanisms governing their function during inflammation and opening new avenues for modulating inflammatory responses.

    1. Cell Biology
    2. Genetics and Genomics
    Keva Li, Nicholas Tolman ... UK Biobank Eye and Vision Consortium
    Research Article

    A glaucoma polygenic risk score (PRS) can effectively identify disease risk, but some individuals with high PRS do not develop glaucoma. Factors contributing to this resilience remain unclear. Using 4,658 glaucoma cases and 113,040 controls in a cross-sectional study of the UK Biobank, we investigated whether plasma metabolites enhanced glaucoma prediction and if a metabolomic signature of resilience in high-genetic-risk individuals existed. Logistic regression models incorporating 168 NMR-based metabolites into PRS-based glaucoma assessments were developed, with multiple comparison corrections applied. While metabolites weakly predicted glaucoma (Area Under the Curve = 0.579), they offered marginal prediction improvement in PRS-only-based models (p=0.004). We identified a metabolomic signature associated with resilience in the top glaucoma PRS decile, with elevated glycolysis-related metabolites—lactate (p=8.8E-12), pyruvate (p=1.9E-10), and citrate (p=0.02)—linked to reduced glaucoma prevalence. These metabolites combined significantly modified the PRS-glaucoma relationship (Pinteraction = 0.011). Higher total resilience metabolite levels within the highest PRS quartile corresponded to lower glaucoma prevalence (Odds Ratiohighest vs. lowest total resilience metabolite quartile=0.71, 95% Confidence Interval = 0.64–0.80). As pyruvate is a foundational metabolite linking glycolysis to tricarboxylic acid cycle metabolism and ATP generation, we pursued experimental validation for this putative resilience biomarker in a human-relevant Mus musculus glaucoma model. Dietary pyruvate mitigated elevated intraocular pressure (p=0.002) and optic nerve damage (p<0.0003) in Lmx1bV265D mice. These findings highlight the protective role of pyruvate-related metabolism against glaucoma and suggest potential avenues for therapeutic intervention.