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Journal of Leukocyte Biology, Vol 61, Issue 2 179-187, Copyright © 1997 by Society for Leukocyte Biology
JOURNAL ARTICLE |
K Konstantopoulos, S Kukreti, CW Smith and LV McIntire
The Cox Laboratory for Biomedical Engineering, Institute of Biosciences and Bioengineering, Rice University, Houston, Texas 77251-1892, USA.
This study demonstrates that endothelial P-selectin and vascular cell adhesion molecule 1 (VCAM-1), but not intercellular adhesion molecule 1 (ICAM-1), are capable of supporting extensive primary adhesion of T cells under flow. To address this issue, we used human umbilical vein endothelial cells (HUVECs) stimulated with histamine, interleukin-4 (IL-4), or interferon-gamma (IFN-gamma) that provide essentially a P-selectin, VCAM-1, or ICAM-1 surface, respectively, in a physiologically relevant cell type. Monoclonal antibody (mAb) blockade studies were carried out to confirm the specificity of these adhesive interactions and rule out a number of other potentially important adhesion molecules. Quantitation of adhesion showed that almost all of the interacting T cells rolled on histamine-stimulated HUVECs or CHO-P cell monolayers. In contrast, approximately 20% of the total interacting T cells with 24-h IL-4-treated HUVECs were firmly adherent. mAb blocking experiments revealed that T cell adhesion to IL-4-treated HUVECs is alpha 4-VCAM-1 dependent. Furthermore, mAb 4B9 directed against domain 1 of VCAM-1 eliminated adhesion, suggesting that alpha 4 integrins may not interact with either the alternatively spliced domain 4 of VCAM-1 or fibronectin in this process. At a wall shear stress of 2 dyn/cm2, the mean T cell rolling velocities were significantly lower on 24-h IL-4-activated HUVECs (10.2 +/- 2.6 microgm/s) compared with either CHO-P cells (15.6 +/- 3.1 microm/s) or histamine-stimulated HUVECs (16.6 +/- 6.1 microm/s). ICAM-1, expressed on the surface of 24-h IFN-gamma-activated HUVECs pretreated with an anti-VCAM-1 mAb to eliminate any VCAM-1-dependent contribution, did not support T cell adhesion under shear conditions. Together these data indicate that T cell primary adhesion can be mediate d by both endothelial P-selectin and VCAM-1 but not ICAM-1. alpha 4 integrins are highly versatile molecules, capable of initiating T cell rolling interactions and mediating firm arrest on activated endothelium.
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