Novel α4-integrin ligands on an endothelial cell line

Tudor, Kim Sue R S; Deem, Tracy L.; Cook‐Mills, Joan M.

doi:10.1139/bcb-78-2-99

Cited by 18 publications

(34 citation statements)

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“…We have reported that lymphocytes bind and migrate across monolayers of the endothelial cell lines, mHEVa and mHEVc (11). We have also previously determined by Ab inhibition studies that adhesion of resting lymphocytes to these endothelial cell lines is dependent on lymphocyte ␣ 4 integrin binding to VCAM-1 on the mHEV cells (12). Therefore, we determined whether adhesion via ␣ 4 integrin to VCAM-1 was required for lymphocyte migration.…”

Section: Lymphocyte Adhesion Is Required For Lymphocyte Migration Acrmentioning

confidence: 89%

“…During lymphocyte migration across the mHEV cell lines, the endothelial cells change shape by retracting their membrane at the site of the lymphocyte, the lymphocyte migrates between adjacent endothelial cells within a few minutes, and the endothelial cells reform their cell-cell junction (11). The mechanism for lymphocyte adhesion to the mHEV cell lines is by lymphocyte ␣ 4 integrin binding to VCAM-1 on the mHEV cells and not to any other known adhesion molecule (12). The expression of VCAM-1 by the mHEV cell lines is constitutive (11), in contrast to other endothelial cells that require cytokine activation of VCAM-1 expression.…”

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confidence: 99%

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Lymphocyte Migration Through Monolayers of Endothelial Cell Lines Involves VCAM-1 Signaling Via Endothelial Cell NADPH Oxidase

Matheny

Deem

Cook‐Mills

2000

The Journal of Immunology

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152

329

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Lymphocytes migrate from the blood across endothelial cells to reach foreign substances sequestered in peripheral lymphoid organs and inflammatory sites. To study intracellular signaling in endothelial cells during lymphocyte migration, we used murine endothelial cell lines that promote lymphocyte migration and constitutively express VCAM-1. The maximum rate of resting splenic lymphocyte migration across monolayers of the endothelial cells occurred at 0–24 h. This migration was inhibited by anti-VCAM-1 or anti-α4 integrin, suggesting that VCAM-1 adhesion was required for migration. To determine whether signals within the endothelial cells were required for migration, irreversible inhibitors of signal transduction molecules were used to pretreat the endothelial cell lines. Inhibitors of NADPH oxidase activity (diphenyleneiodonium and apocynin) blocked migration >65% without affecting adhesion. Because NADPH oxidase catalyzes the production of reactive oxygen species (ROS), we examined whether ROS were required for migration. Scavengers of ROS inhibited migration without affecting adhesion. Furthermore, VCAM-1 ligand binding stimulated NADPH oxidase-dependent production of ROS by the endothelial cells lines and primary endothelial cell cultures. Finally, VCAM-1 ligand binding induced an apocynin-inhibitable actin restructuring in the endothelial cell lines at the location of the lymphocyte or anti-VCAM-1-coated bead, suggesting that an NADPH oxidase-dependent endothelial cell shape change was required for lymphocyte migration. In summary, VCAM-1 signaled the activation of endothelial cell NADPH oxidase, which was required for lymphocyte migration. This suggests that endothelial cells are not only a scaffold for lymphocyte adhesion, but play an active role in promoting lymphocyte migration.

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Section: Lymphocyte Adhesion Is Required For Lymphocyte Migration Acrmentioning

confidence: 89%

mentioning

confidence: 99%

Lymphocyte Migration Through Monolayers of Endothelial Cell Lines Involves VCAM-1 Signaling Via Endothelial Cell NADPH Oxidase

Matheny

Deem

Cook‐Mills

2000

The Journal of Immunology

Self Cite

152

329

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“…B6RV2 cells were maintained in RPMI with 15% FBS. Murine ECs, mHEVc, were cultured as described (Cook-Mills et al 1996;Tudor et al 2000).…”

Section: Cell Lines and Growth Conditionsmentioning

confidence: 99%

Bone marrow-derived endothelial progenitor cells are a major determinant of nascent tumor neovascularization

Nolan¹,

Ciarrocchi²,

Mellick³

et al. 2007

Genes Dev.

366

306

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Tumors build vessels by cooption of pre-existing vasculature and de novo recruitment of bone marrow (BM)-derived endothelial progenitor cells (EPCs). However, the contribution and the functional role of EPCs in tumor neoangiogenesis are controversial. Therefore, by using genetically marked BM progenitor cells, we demonstrate the precise spatial and temporal contribution of EPCs to the neovascularization of three transplanted and one spontaneous breast tumor in vivo using high-resolution microscopy and flow cytometry. We show that early tumors recruit BM-derived EPCs that differentiate into mature BM-derived endothelial cells (ECs) and luminally incorporate into a subset of sprouting tumor neovessels. Notably, in later tumors, these BM-derived vessels are diluted with non-BM-derived vessels from the periphery, which accounts for purported differences in previously published reports. Furthermore, we show that specific ablation of BM-derived EPCs with ␣-particle-emitting anti-VE-cadherin antibody markedly impaired tumor growth associated with reduced vascularization. Our results demonstrate that BM-derived EPCs are critical components of the earliest phases of tumor neoangiogenesis.[Keywords: Bone marrow transplantation; tumor angiogenesis; endothelial progenitor cells; endothelial cells; VE-cadherin; neovascularization] Supplemental material is available at http://www.genesdev.org.

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“…Therefore, spleen cells (3 ϫ 10 6 ) were added to the flow chamber (3.5 cm 2 ) at 2 dynes/cm 2 . Spleen cells were added at concentrations below saturation of monolayer cell binding or migration (12,24). To initiate spleen cell contact with the endothelial cells in vitro, the spleen cells were allowed to settle in the chamber as monitored by microscopy and then 2 dynes/cm 2 was applied for the 15 min laminar flow assay.…”

Section: Cell Association and Migration Assays With Laminar Flowmentioning

confidence: 99%

“…The endothelial cell lines, mHEVa and mHEVc, constitutively express VCAM-1 and do not express other ligands for lymphocyte migration such as ICAM-1, PECAM-1, and E-selectin as determined by immunofluorescence labeling (24) or cDNA microarray analysis (data not shown). Furthermore, adhesion and migration across the endothelial cells are dependent on binding to VCAM-1 and the constitutive expression of MCP-1 by the endothelial cells (9,24,25). Abs for other adhesion molecules other than VCAM-1 or its ligand ␣ 4 ␤ 1 integrin do not block lymphocyte migration across the endothelial cells (12).…”

mentioning

confidence: 98%

VCAM-1 Activation of Endothelial Cell Protein Tyrosine Phosphatase 1B

Deem

Abdala‐Valencia

Cook‐Mills

2007

The Journal of Immunology

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Lymphocytes migrate from the blood into tissue by binding to and migrating across endothelial cells. One of the endothelial cell adhesion molecules that mediate lymphocyte binding is VCAM-1. We have reported that binding to VCAM-1 activates endothelial cell NADPH oxidase for the generation of reactive oxygen species (ROS). The ROS oxidize and stimulate an increase in protein kinase C (PKC)α activity. Furthermore, these signals are required for VCAM-1-dependent lymphocyte migration. In this report, we identify a role for protein tyrosine phosphatase 1B (PTP1B) in the VCAM-1 signaling pathway. In primary cultures of endothelial cells and endothelial cell lines, Ab cross-linking of VCAM-1 stimulated an increase in serine phosphorylation of PTP1B, the active form of PTP1B. Ab cross-linking of VCAM-1 also increased activity of PTP1B. This activation of PTP1B was downstream of NADPH oxidase and PKCα in the VCAM-1 signaling pathway as determined with pharmacological inhibitors and antisense approaches. In addition, during VCAM-1 signaling, ROS did not oxidize endothelial cell PTP1B. Instead PTP1B was activated by serine phosphorylation. Importantly, inhibition of PTP1B activity blocked VCAM-1-dependent lymphocyte migration across endothelial cells. In summary, VCAM-1 activates endothelial cell NADPH oxidase to generate ROS, resulting in oxidative activation of PKCα and then serine phosphorylation of PTP1B. This PTP1B activity is necessary for VCAM-1-dependent transendothelial lymphocyte migration. These data show, for the first time, a function for PTP1B in VCAM-1-dependent lymphocyte migration.

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Novel α4-integrin ligands on an endothelial cell line

Cited by 18 publications

References 0 publications

Lymphocyte Migration Through Monolayers of Endothelial Cell Lines Involves VCAM-1 Signaling Via Endothelial Cell NADPH Oxidase

Lymphocyte Migration Through Monolayers of Endothelial Cell Lines Involves VCAM-1 Signaling Via Endothelial Cell NADPH Oxidase

Bone marrow-derived endothelial progenitor cells are a major determinant of nascent tumor neovascularization

VCAM-1 Activation of Endothelial Cell Protein Tyrosine Phosphatase 1B

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