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Endothelial basement membrane laminin α5 selectively inhibits T lymphocyte extravasation into the brain

Nature Medicine volume 15, pages 519–527 (2009)Cite this article

Abstract

Specific inhibition of the entry of encephalitogenic T lymphocytes into the central nervous system in multiple sclerosis would provide a means of inhibiting disease without compromising innate immune responses. We show here that targeting lymphocyte interactions with endothelial basement membrane laminins provides such a possibility. In mouse experimental autoimmune encephalomyelitis, T lymphocyte extravasation correlates with sites expressing laminin α4 and small amounts of laminin α5. In mice lacking laminin α4, laminin α5 is ubiquitously expressed along the vascular tree, resulting in marked and selective reduction of T lymphocyte infiltration into the brain and reduced disease susceptibility and severity. Vessel phenotype and immune response were not affected in these mice. Rather, laminin α5 directly inhibited integrin α6β1–mediated migration of T lymphocytes through laminin α4. The data indicate that T lymphocytes use mechanisms distinct from other immune cells to penetrate the endothelial basement membrane barrier, permitting specific targeting of this immune cell population.

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Figure 1: Immunofluorescence and electron microscopy analyses of WT and Lama4−/− brains.
Figure 2: Active EAE induction in Lama4−/− mice and WT littermates.
Figure 3: Passive EAE experiments.
Figure 4: T lymphocyte proliferation studies.
Figure 5: Encephalitogenic T lymphocyte transmigration studies.
Figure 6: Active EAE induction in WT mice lacking functional laminin α4 receptor, integrin α6β1.

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Acknowledgements

This work was supported by the German (SFB293 A14, B8, A1; SFB492 Z3) and Swedish Research Councils (K2005-06X-14184-04A, 621-2001-2142), Alfred Österlunds Foundation, Knut and Alice Wallenbergs Foundation (KAW 2002.0056), the Crafoord Foundation, the Greta and Johan Kocks Foundation and the Interdisciplinary Clinical Research Center (IZKF; Lo2/017/07) in Münster, Germany. We thank M. Sixt for initial studies on Lama4−/− mice, J. Eble (Frankfurt University) for recombinant integrin α6β1, A. Sonnenberg (Division of Cell Biology, The Netherlands Cancer Institute) for GoH3, A. De Arcangelis for tissue collection, G. Roos for technical assistance and F. Kiefer and R. Böhmer for assistance with confocal microscopy.

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Author notes

  1. Per Anderson

    Present address: Current address: Instituto de Parasitología y Biomedicina, Parque Tecnológico de Ciencias de la Salud, Armilla (Granada), Spain.,

Authors and Affiliations

  1. Institute for Physiological Chemistry and Pathobiochemistry, Münster University, Münster, Germany

    Chuan Wu, Rupert Hallmann, Jian Song, Eva Korpos & Lydia M Sorokin

  2. Department of Experimental Medical Science, Section for Immunology, Lund University, Lund, Sweden

    Fredrik Ivars & Per Anderson

  3. Max Planck Institute for Molecular Biomedicine, Vascular Cell Biology, Müenster, Germany

    Dietmar Vestweber

  4. Radiation Physics, Lund University, Lund, Sweden

    Per Nilsson

  5. Leibniz Institute for Arterosclerosis Research, Münster University, Münster, Germany

    Horst Robenek

  6. Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden

    Karl Tryggvason

  7. Department of Dermatology, Münster University, Münster, Germany

    Karin Loser & Stefan Beissert

  8. Institut de Génétique et de Biologie Moléculaire et Cellulaire, Strasbourg, France

    Elisabeth Georges-Labouesse

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Contributions

All experimental work was carried out by C.W. P.A. and F.I. contributed to the in vivo and in vitro T lymphocyte proliferation studies; P.N. was instrumental in generation of bone marrow–chimeric mice; H.R. carried out all electron microscopy studies; the Lama4−/− mouse was generated in K.T.'s laboratory; R.H. was instrumental in project development and experimental design; D.V. provided expertise and tools for assessment of endothelial cell-to-cell contacts in Lama4−/− mice; K.L., S.B. and J.S. provided expertise and tools for FACS analyses; E.K. carried out immunofluorescence analyses; E.G.-L. provided the Itga6−/− embryonic liver cells for generation of bone marrow–chimeric mice; project development and all experimental work was carried out under the supervision and in the laboratory of L.M.S.

Corresponding author

Correspondence to Lydia M Sorokin.

Supplementary information

Supplementary Text and Figures

Supplementary Figs. 1–5 and Supplementary Methods (PDF 5107 kb)

Supplementary Video 1

Quick time movie of 0.4-μm Z stacks through a portion of the 60-μm section of WT mouse brain double-stained for laminin α4 (green) and laminin α5 (red) shown in Figure 1a. (MOV 2363 kb)

Supplementary Video 2

Quick time movie of 0.4-μm Z stacks through a portion of the 60-μm section of WT EAE brain double-stained for laminin α5 (green) and CD45 (red) shown in Figure 1b. (MOV 4141 kb)

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Wu, C., Ivars, F., Anderson, P. et al. Endothelial basement membrane laminin α5 selectively inhibits T lymphocyte extravasation into the brain. Nat Med 15, 519–527 (2009). https://doi.org/10.1038/nm.1957

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