The sphingosine 1-phosphate receptor 1 causes tissue retention by inhibiting the entry of peripheral tissue T lymphocytes into afferent lymphatics

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Although much is known about the migration of T cells from blood to lymph nodes, less is known about the mechanisms regulating the migration of T cells from tissues into lymph nodes through afferent lymphatics. Here we investigated T cell egress from nonlymphoid tissues into afferent lymph in vivo and developed an experimental model to recapitulate this process in vitro. Agonism of sphingosine 1-phosphate receptor 1 inhibited the entry of tissue T cells into afferent lymphatics in homeostatic and inflammatory conditions and caused the arrest, mediated at least partially by interactions of the integrin LFA-1 with its ligand ICAM-1 and of the integrin VLA-4 with its ligand VCAM-1, of polarized T cells at the basal surface of lymphatic but not blood vessel endothelium. Thus, the increased sphingosine 1-phosphate present in inflamed peripheral tissues may induce T cell retention and suppress T cell egress.

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Figure 1: Chemokine and adhesion receptors used for entry into lymph nodes through blood or afferent lymphatics.
Figure 2: S1P receptor stimulation inhibits migration to lymph nodes through afferent lymphatics but not through blood.
Figure 3: Tissue S1P and lymphocyte S1P receptor in the suppression of afferent lymphatic migration during acute inflammation.
Figure 4: SVEC4-10 cells express lymphatic lineage markers.
Figure 5: Migration of T cells across SVEC4-10 cells is polarized from the basal to the apical surface and is dependent on LFA-1.
Figure 6: T cell migration across lymphatic but not blood vascular endothelium is regulated by S1P1.
Figure 7: FTY720 inhibits the motility of T cells across endothelial cell layers.
Figure 8: S1P receptor stimulation causes arrest of the migration of T cells at the basal surface of the lymphatic endothelium.
Figure 9: T cells with stimulated S1P receptors block the migration of untreated T cells and adhere to lymphatic endothelium.

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We thank P. Heeger and R. Jessberger for discussions; V. Brinkmann for reagents; J. Lowe (Cleveland Clinic) for mice lacking fucosyltransferase IV and VII; R. Proia (National Institutes of Health) for S1P1loxP/loxP Lck-Cre mice and littermate controls; and M. Mao, D. Chen, A.M. Ledgerwood, the Mount Sinai Microscopy Core and the Mount Sinai Microarray Facility. Supported by the National Institutes of Health (AI41428 and AI62765 to J.S.B., and DK67381 to S.A.L.), the Emerald Foundation (J.S.B.), the Juvenile Diabetes Research Foundation (1-2005-16 to J.S.B.), Ministerio de Educación y Ciencia, Spain (SAF2007-63579 to J.C.O.) and the Howard Hughes Medical Institute (L.G.L.).

Author information

L.G.L., G.L. and J.C.O. planned, did and analyzed experiments and prepared the manuscript; N.Z. did experiments analyzing in vivo migration; A.G. and S.A.L. planned and did RT-PCR for endothelial cell line characterization; S.J.E., F.G., M.M., H.P., Y.D. and Y.Y. planned and did various aspects of in vivo and in vitro experiments; X.H. and E.H.S. did experiments to measure tissue lipid concentrations; M.L.A. designed S1P1loxP/loxP Lck-Cre transgenic mice; and J.S.B. planned and analyzed experiments and prepared the manuscript.

Correspondence to Jonathan S Bromberg.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–5, Table 2 and Methods (PDF 593 kb)

Supplementary Table 1 (XLS 7257 kb)

Supplementary Video 1

Time-lapse video of untreated T cells migrating across iSVEC4-10. Colored lines represent paths of individual cells over the course of the experiment. (AVI 2499 kb)

Supplementary Video 2

Time-lapse video of FTY720-treated T cells migrating across iSVEC4-10. Colored lines represent paths of individual cells over the course of the experiment. (AVI 2448 kb)

Supplementary Video 3

Three-dimensional rendering of Z-stack images from untreated T cells migrating across an iSVEC4-10 endothelial layer. T cells (green) were stained with CFSE, endothelial cells (red) stained with PKH26, and slides were mounted with DAPI-containing mounting solution (blue). (AVI 2579 kb)

Supplementary Video 4

Three-dimensional rendering of Z-stack images from FTY720-treated T cells migrating across an iSVEC4-10 endothelial layer. T cells (green) were stained with CFSE, endothelial cells (red) were stained with PKH26, and slides were mounted with DAPI-containing mounting solution (blue). (AVI 9888 kb)

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Ledgerwood, L., Lal, G., Zhang, N. et al. The sphingosine 1-phosphate receptor 1 causes tissue retention by inhibiting the entry of peripheral tissue T lymphocytes into afferent lymphatics. Nat Immunol 9, 42–53 (2008) doi:10.1038/ni1534

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