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J. Exp. Med.,
Volume 186, Number 10, November 17, 1997 1701-1711
By
From Veterinary Molecular Biology, Montana State University, Bozeman, Montana 59717
Bovine 
/
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 / 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 / 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 / T
cells in a mixed lymphocyte preparation. GR113 mAb blocked the binding of the soluble
platelet molecule to the / 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.
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