Determination of T-lymphocyte adhesion to endothelial cells using a novel luminescence marker.
Here we report the quantification of T cell adhesion to endothelial cells using the luminescence marker BioLite. A new application for this substance was established using a microassay for the detection of TK-1 mouse T-lymphoma cell adhesion to unstimulated or TNF alpha-stimulated eEnd.2 mouse vascular endothelioma cells. Prelabelling of TK-1 with the marker resulted in luminescence values linearly related to the number of adherent T cells. The marker was not toxic for T cells or endothelial cells nor did it interfere with the expression or function of adhesion molecules on T cells (LFA-1, VLA-4) or endothelial cells (ICAM-1, VCAM-1). When compared with established techniques to quantify T cell/endothelial cell adhesion, i.e. microscopical evaluation or isotope prelabelling of T cells, this method was found to be just as reliable and sensitive.
Termeer
C C
CC
Department of Dermatology, University of Freiburg, Germany.
Weiss
J M
JM
Dittmar
H
H
Vanscheidt
W
W
Schöpf
E
E
Simon
J C
JC
D000818:Animals; D002244:Carbon; D002448:Cell Adhesion; D002460:Cell Line; D002465:Cell Movement / Q000276:immunology*; D004730:Endothelium, Vascular / Q000166:cytology* / Q000276:immunology; D018799:Intercellular Adhesion Molecule-1 / Q000276:immunology; D008163:Luminescent Measurements; D051379:Mice; D013601:T-Lymphocytes / Q000166:cytology* / Q000276:immunology; D019010:Vascular Cell Adhesion Molecule-1 / Q000276:immunology
C039468:Biolite; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; D002244:Carbon