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Shear forces promote lymphocyte migration across vascular endothelium bearing apical chemokines

Nature Immunology volume 2, pages 515–522 (2001)Cite this article

Abstract

Leukocyte transendothelial migration (TEM) is thought to be a chemotactic process controlled by chemokine gradients across the endothelium. Using cytokine-activated human umbilical vascular endothelial cells (HUVECs) as a model of inflamed endothelium, we have shown that apical endothelial chemokines can trigger robust peripheral blood lymphocyte (PBL) migration across endothelial cells. Lymphocyte TEM was promoted by physiological shear stress applied continuously to migrating lymphocytes. Lymphocyte integrins, intact actin cytoskeleton and Gi protein–mediated chemokine signaling, but not a chemotactic gradient, were mandatory for TEM. PBL TEM did not require intracellular free calcium or intact phosphatidyl inositol kinase activity in migrating lymphocytes. Thus, lymphocyte TEM is promoted by fluid shear-induced mechanical signals coupled to Gi protein signals at apical endothelial zones.

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Figure 1: Analysis of leukocyte TEM under physiological shear flow in a parallel plate flow-chamber assay.
Figure 2: Apical endothelial chemokines promote lymphocyte TEM under physiological flow conditions.
Figure 3: Role of integrin avidity in PBL adhesion and TEM through TNF-α–activated HUVECs under physiological flow.
Figure 4: Shear stress must be continuously applied on adherent lymphocytes rather than on the endothelial surface to promote TEM.
Figure 5: Ultrastructural analysis of lymphocyte-endothelium contacts under different shear flow conditions.
Figure 6: The context and dose-dependence of chemokine signaling to PBLs are critical for transmigration across TNF-α–activated HUVECs.
Figure 7: Chemokine-triggered lymphocyte TEM does not depend on the integrity of the endothelial barrier junctions.
Figure 8: Chemokine-triggered lymphocyte TEM does not require PI3K activity or intracellular Ca2+.

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Acknowledgements

We thank M. Lipp, R. Lobb and S. Raffi, for providing reagents and S. Shwarzbaum for editorial help. Special thanks to S. Feigelson and A. Peled for helpful discussions. Supported in part by the Israel Science Foundation and the Minnerva Foundation, Germany.

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  1. Department of Immunology, The Weizmann Institute of Science, Rehovot, 76100, Israel

    Guy Cinamon, Vera Shinder & Ronen Alon

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Correspondence to Ronen Alon.

Supplementary information

Web Movie 1.

Digitized Quicktime videos showing characteristic scenes from time-lapse recordings of lymphocytes accumulated on TNF-α-activated HUVEC monolayers overlaid with SDF-1a (100 ng/ml) and subjected to continuous shear stress of 5 dyn/cm2. The time (in min) elapsed from the beginning of the shear application phase is shown at the top. Shown is a magnified image of a 290x200 micron field, selected from full 540x400 micron field (described in Methods). Lymphocytes are numbered from 1–10 to allow monitoring of their position throughout the assay. Arrows point to cells 6 and 8, which are in the process of TEM. Lymphocyte number 7 undergoes TEM at t=2.55 min. The remaining lymphocytes locomote over the surface without transmigrating through the ECs. (MOV 5476 kb)

Web Movie 2.

Digitized Quicktime videos showing characteristic scenes from timelapse recordings of lymphocytes accumulated on TNF-α-activated HUVEC monolayers overlaid with SDF-1α (100 ng/ml) and subjected to shear-free conditions. The time (in min) elapsed from the beginning of the shear application phase is shown at the top. For further details refer to the legend of Web Movie 1. No lymphocyte TEM occurs under these experimental conditions. (MOV 5306 kb)

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Cinamon, G., Shinder, V. & Alon, R. Shear forces promote lymphocyte migration across vascular endothelium bearing apical chemokines. Nat Immunol 2, 515–522 (2001). https://doi.org/10.1038/88710

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