title: Endothelial P-selectin and VCAM-1 each can function as primary adhesive mechanisms for T cells under conditions of flow.
abstract: 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.
authors.affiliation: The Cox Laboratory for Biomedical Engineering, Institute of Biosciences and Bioengineering, Rice University, Houston, Texas 77251-1892, USA.
authors.forename: K
authors.initials: K
authors.lastname: Konstantopoulos
authors.forename: S
authors.initials: S
authors.lastname: Kukreti
authors.forename: C W
authors.initials: CW
authors.lastname: Smith
authors.forename: L V
authors.initials: LV
authors.lastname: McIntire
mesh_terms: D000818:Animals; D001783:Blood Flow Velocity; D016466:CHO Cells; D002448:Cell Adhesion / Q000187:drug effects / Q000276:immunology*; D006224:Cricetinae; D004730:Endothelium, Vascular / Q000187:drug effects / Q000276:immunology / Q000502:physiology*; D005434:Flow Cytometry; D018056:Hemorheology; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D015847:Interleukin-4 / Q000494:pharmacology; D019007:P-Selectin / Q000502:physiology*; D013601:T-Lymphocytes / Q000187:drug effects / Q000502:physiology*; D014471:Umbilical Veins; D019010:Vascular Cell Adhesion Molecule-1 / Q000502:physiology*
chemical_list: D019007:P-Selectin; D019010:Vascular Cell Adhesion Molecule-1; D015847:Interleukin-4; D007371:Interferon-gamma