The active role of the endothelium in acute inflammatory processes has only recently been appreciated. In particular, the identification and molecular cloning of the cytokine regulated endothelial-leukocyte adhesion molecules, ELAM-1 and ICAM-1, provides a basis to understand the cellular mechanisms involved in leukocyte-endothelial interactions during inflammation. Preliminary studies by the applicant have demonstrated that cytokine- activated endothelial monolayers promote an upregulated neutrophil transmigration and that expression of both ELAM-1 and ICAM-1 are required for this process. This project will examine the cellular and molecular mechanisms of neutrophil activation during their adhesive interactions with cytokine activated endothelium, focusing on the role of ELAM-1. The primary goals of this project are to: determine whether neutrophils become activated during adhesive interactions with cytokine-activated endothelial monolayers as measured by biochemical (e.g., mobilization of cytosolic Ca2+ by microspectrofluorimetry) and morphological (time lapse videomicroscopy) parameters; 2) to determine whether purified ELAM-1 (human recombinant ELAM-1) deposited on inert surfaces can directly activate neutrophils using the same indices as above; 3) to determine which epitopes on ELAM-1 are involved in neutrophil activation using monoclonal antibodies which recognize various epitopes on ELAM-1; 4) using an in vitro transmigration assay, assess the role of ELAM-1-dependent neutrophil activation in neutrophil activation. Thus, the proposed studies will combine cell biological techniques and immunological approaches to gain insight into endothelial-dependent mechanisms important in the local regulation of inflammation, i.e., neutrophil adhesion and transmigration.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
First Independent Research Support & Transition (FIRST) Awards (R29)
Project #
5R29HL047646-03
Application #
2223827
Study Section
Pathology A Study Section (PTHA)
Project Start
1992-02-01
Project End
1997-01-31
Budget Start
1994-02-01
Budget End
1995-01-31
Support Year
3
Fiscal Year
1994
Total Cost
Indirect Cost
Name
Brigham and Women's Hospital
Department
Type
DUNS #
071723621
City
Boston
State
MA
Country
United States
Zip Code
02115
Goetz, D J; Greif, D M; Shen, J et al. (1999) Cell-cell adhesive interactions in an in vitro flow chamber. Methods Mol Biol 96:137-45
Allport, J R; Ding, H; Collins, T et al. (1997) Endothelial-dependent mechanisms regulate leukocyte transmigration: a process involving the proteasome and disruption of the vascular endothelial-cadherin complex at endothelial cell-to-cell junctions. J Exp Med 186:517-27
Montoya, M C; Luscinskas, F W; del Pozo, M A et al. (1997) Reduced intracellular oxidative metabolism promotes firm adhesion of human polymorphonuclear leukocytes to vascular endothelium under flow conditions. Eur J Immunol 27:1942-51
Allport, J R; Ding, H T; Ager, A et al. (1997) L-selectin shedding does not regulate human neutrophil attachment, rolling, or transmigration across human vascular endothelium in vitro. J Immunol 158:4365-72
Luscinskas, F W; Ding, H; Tan, P et al. (1996) L- and P-selectins, but not CD49d (VLA-4) integrins, mediate monocyte initial attachment to TNF-alpha-activated vascular endothelium under flow in vitro. J Immunol 157:326-35
Luscinskas, F W; Gimbrone Jr, M A (1996) Endothelial-dependent mechanisms in chronic inflammatory leukocyte recruitment. Annu Rev Med 47:413-21
Pierce, J W; Read, M A; Ding, H et al. (1996) Salicylates inhibit I kappa B-alpha phosphorylation, endothelial-leukocyte adhesion molecule expression, and neutrophil transmigration. J Immunol 156:3961-9
Goetz, D J; Ding, H; Atkinson, W J et al. (1996) A human colon carcinoma cell line exhibits adhesive interactions with P-selectin under fluid flow via a PSGL-1-independent mechanism. Am J Pathol 149:1661-73
Gerritsen, M E; Shen, C P; McHugh, M C et al. (1995) Activation-dependent isolation and culture of murine pulmonary microvascular endothelium. Microcirculation 2:151-63
Read, M A; Neish, A S; Luscinskas, F W et al. (1995) The proteasome pathway is required for cytokine-induced endothelial-leukocyte adhesion molecule expression. Immunity 2:493-506

Showing the most recent 10 out of 14 publications