Letter | Published:

Structural and functional features of central nervous system lymphatic vessels

Nature volume 523, pages 337341 (16 July 2015) | Download Citation

  • A Corrigendum to this article was published on 24 February 2016

Abstract

One of the characteristics of the central nervous system is the lack of a classical lymphatic drainage system. Although it is now accepted that the central nervous system undergoes constant immune surveillance that takes place within the meningeal compartment1,2,3, the mechanisms governing the entrance and exit of immune cells from the central nervous system remain poorly understood4,5,6. In searching for T-cell gateways into and out of the meninges, we discovered functional lymphatic vessels lining the dural sinuses. These structures express all of the molecular hallmarks of lymphatic endothelial cells, are able to carry both fluid and immune cells from the cerebrospinal fluid, and are connected to the deep cervical lymph nodes. The unique location of these vessels may have impeded their discovery to date, thereby contributing to the long-held concept of the absence of lymphatic vasculature in the central nervous system. The discovery of the central nervous system lymphatic system may call for a reassessment of basic assumptions in neuroimmunology and sheds new light on the aetiology of neuroinflammatory and neurodegenerative diseases associated with immune system dysfunction.

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Acknowledgements

We thank S. Smith for editing the manuscript; V. Engelhard for initial discussions and suggestions on lymphatic endothelial cell specific markers. We also thank the members of the Kipnis laboratory, Brain Immunology and Glia (BIG) center, and the Department of Neuroscience at the University of Virginia (especially J. Lukens) for their valuable comments during multiple discussions of this work. L. Holland and B. Lopes provided the human samples. This work was funded by “Fondation pour la Recherche Medicale” to A.L. and by The National Institutes of Health (R01AG034113 and R01NS061973) to J.K.

Author information

Affiliations

  1. Center for Brain Immunology and Glia, School of Medicine, University of Virginia, Charlottesville, Virginia 22908, USA

    • Antoine Louveau
    • , Igor Smirnov
    • , Timothy J. Keyes
    • , Noel C. Derecki
    • , Kevin S. Lee
    • , Tajie H. Harris
    •  & Jonathan Kipnis
  2. Department of Neuroscience, School of Medicine, University of Virginia, Charlottesville, Virginia 22908, USA

    • Antoine Louveau
    • , Igor Smirnov
    • , Timothy J. Keyes
    • , Noel C. Derecki
    • , Kevin S. Lee
    • , Tajie H. Harris
    •  & Jonathan Kipnis
  3. Medical Scientist Training Program, School of Medicine, University of Virginia, Charlottesville, Virginia 22908, USA

    • Jacob D. Eccles
    • , Sherin J. Rouhani
    • , J. David Peske
    •  & Jonathan Kipnis
  4. Beirne B. Carter Center for Immunology Research, School of Medicine, University of Virginia, Charlottesville, Virginia 22908, USA

    • Jacob D. Eccles
    • , Sherin J. Rouhani
    •  & J. David Peske
  5. Department of Medicine (Division of Allergy), School of Medicine, University of Virginia, Charlottesville, Virginia 22908, USA

    • Jacob D. Eccles
  6. Department of Microbiology, Immunology, and Cancer Biology, School of Medicine, University of Virginia, Charlottesville, Virginia 22908, USA

    • Sherin J. Rouhani
    •  & J. David Peske
  7. Department of Cell Biology, School of Medicine, University of Virginia, Charlottesville, Virginia 22908, USA

    • David Castle
  8. Department of Pathology (Neuropathology), School of Medicine, University of Virginia, Charlottesville, Virginia 22908, USA

    • James W. Mandell
  9. Department of Neurosurgery, School of Medicine, University of Virginia, Charlottesville, Virginia 22908, USA

    • Kevin S. Lee

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Contributions

A.L. performed most of the experiments, analysed the data, and contributed to experimental design and manuscript writing. I.S. performed all the surgeries and intracerebroventricular injections. T.J.K. assisted with the experiments and the analysis of the data. J.D.E, S.J.R. and J.D.P. participated in the discussions and helped with the experimental design. N.C.D. performed the xDCLN experiment. D.C. contributed to the imaging and the analysis of the electron microscopy images. J.W.M. contributed with data analysis of the human samples. K.S.L. contributed to experimental design and to manuscript editing. T.H.H. assisted to the intravital imaging experiment, contributed to experimental design and to manuscript editing. J.K. designed the study, assisted with data analysis, and wrote the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding authors

Correspondence to Antoine Louveau or Jonathan Kipnis.

Extended data

Supplementary information

Videos

  1. 1.

    3D reconstruction of the meningeal lymphatic vessels around the superior sagittal sinus

    Adult mice were injected i.v. with DyLight 488 lectin prior to euthanasia. Meninges were harvested and stained for lymphatic endothelial cells (Lyve-1). A 3D reconstruction of both the blood vasculature (green) and the meningeal lymphatic vessels (red) using IMARIS software are presented.

  2. 2.

    Morphological features of the meningeal lymphatic vessels

    Meninges and diaphragm from adult mice were harvested and stained for lymphatic endothelial cells (Lyve-1). A 3D reconstruction of the lymphatic network using IMARIS software is presented.

  3. 3.

    CSF-filled vessel lining the dural sinuses

    Adult mice were injected i.v. with fluorescein and i.c.v. with QDot655. The superior sagittal sinus was imaged intravitally by 2-photon microscope under a thinned skull. The CSF-filled vessel (red) is visible between the sinus (left) and the CSF (right).

  4. 4.

    3D reconstruction of the meningeal lymphatic vessel by multiphoton microscopy

    Adult mice were injected i.v. with QDot655 (red) and i.c.v. with Alexa488 conjugated anti-Lyve-1 antibody (green). The superior sagittal sinus was imaged intravitally by 2-photon microscope under a thinned skull and the surface was reconstructed using IMARIS software. The signal from the remaining skull (blue) was recorded.

  5. 5.

    Flow directionality of the meningeal lymphatics

    Adult mice were injected i.v. with fluorescein and i.c.v. with QDot655. The superior sagittal sinus was imaged intravitally by 2-photon microscope under a thinned skull. The blood flow in the sinus (green, left) appears faster than the one in the meningeal lymphatic vessels (red, right). The flow inside of the lymphatic vessels appears unidirectional.

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DOI

https://doi.org/10.1038/nature14432

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