The Journal of Experimental Medicine
Torrey Pines Biolabs
  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents
This Article
Right arrow Full Text
Right arrow Full Text (PDF, 310K)
Right arrow PPT slides of all figures
Right arrow Alert me when this article is cited
Right arrow Citation Map
Services
Right arrow Email this article
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new content in the JEM
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Jutila, M. A.
Right arrow Articles by Kurk, S.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Jutila, M. A.
Right arrow Articles by Kurk, S.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Facebook   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?
© The Rockefeller University Press, 0022-1007/1997/11/1701/ $5.00
The Journal of Experimental Medicine, Volume 186, Number 10, November 17, 1997 1701-1711

Articles

Characterization of an Adhesion Molecule that Mediates Leukocyte Rolling on 24 h Cytokine- or Lipopolysaccharide-stimulated Bovine Endothelial Cells under Flow Conditions

Mark A. Jutila, Eric Wilson, and Sandy Kurk

From Veterinary Molecular Biology, Montana State University, Bozeman, Montana 59717

Bovine {gamma}/{delta} T cells and neutrophils roll on 24 h cytokine- or lipopolysaccharide-stimulated bovine fetal umbilical cord endothelial cells in assays done under physiological flow. An antibody directed against E- and L-selectin has minimal blocking effect on this rolling interaction. mAbs were raised against the stimulated bovine endothelial cells and screened for inhibition of {gamma}/{delta} T cell rolling. One mAb (GR113) was identified that recognizes an antigen (GR antigen) selectively expressed by stimulated bovine endothelial cells isolated from fetal umbilical cord, mesenteric lymph nodes, and aorta. GR113 blocked bovine {gamma}/{delta} T cell as well as neutrophil rolling on the 24 h-activated endothelial cells. The GR antigen was constitutively expressed at low levels on the cell surface of platelets and its expression was not upregulated after stimulation of these cells with thrombin or phorbol myristate acetate. However, stimulated platelets released a soluble, functionally active form of the molecule that selectively bound in solution to {gamma}/{delta} T cells in a mixed lymphocyte preparation. GR113 mAb blocked the binding of the soluble platelet molecule to the {gamma}/{delta} T cells. Soluble GR antigen also bound a subset of human lymphocytes. Cutaneous lymphocyte-associated antigen (CLA) bright human lymphocytes exhibited the greatest capacity to bind the GR antigen, though CLA was not required for binding. Subsets of both human CD4 and CD8 T cells bound the GR antigen. Immunoprecipitation experiments showed the GR antigen to be 110-120 kD Mr. The binding of soluble GR antigen was inhibited by EDTA and O-sialoglycoprotease, but not neuraminidase treatment of the target cells.

Address correspondence to Mark Jutila, Veterinary Molecular Biology, Montana State University, Bozeman, MT 59717. Phone: 406-994-4705; FAX: 406-994-4303; E-mail: uvsmj{at}gemini.oscs.montana.edu

Eugene Butcher, Ward Jones, and Kemal Aydintug are thanked for constructive comments and useful discussions. The authors also thank Dr. Joyce Bischoff (Harvard University) for the polyclonal antibodies against bovine E- and P-selectin, and helpful comments on our experiments and this manuscript.

This work was funded by grants from USDA NRI 96-35204-3580, Animal Health, and the Montana Agricultural Experiment Station (J-5150).

1 Abbreviations used in this paper: BUVECs, bovine umbilical vein endothelial cells; CLA, cutaneous lymphocyte-associated antigen; MAdCAM-1, mucosal addressin cell adhesion molecule-1; PSGL-1, P-selectin glycoprotein ligand-1; PVDF, polyvinylidene difluoride; VCAM, vascular cell adhesion molecule-1.


Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Facebook Facebook   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?


This article has been cited by other articles:



  Home | Help | Feedback | Subscriptions | Archive | Search
TABLE OF CONTENTS