A method for measuring leukocyte rolling on the selectins

Analytical Biochemistry
J J Schmuke, J K Welply

Abstract

An inexpensive, high-throughput method to simulate leukocyte rolling in the microvasculature has been developed. The method utilizes a 0.22-mm-inner diameter, fused silica capillary tube, coated with E- or P-selectin. Fluorescently labeled HL-60 cells are delivered to the capillary tube at a constant flow rate, exposing the cells to wall stresses approximating those found in postcapillary venules. Cells that physically associate with the inner walls of the tube and whose rate of movement through the tube is retarded, i.e., rolling cells, are monitored by fluorescence microscopy. Images are recorded on a time-lapse videocassette recorder. Both rolling incidence and velocity were shown to be related to the concentration of selectin utilized to coat the tube. Due to the extremely small volume (50 microliters) required to fill the capillary tube, this technique is useful for testing the effect of limited quantities of potential antagonists on cell rolling. Using this technique, sLex(Glc) tetrasaccharide was shown to prevent the rolling of HL-60 cells on immobilized E-selectin while fucoidan and dextran sulfate were shown to inhibit rolling of HL-60 cells on P-selectin.

Citations

Feb 19, 2004·Advanced Drug Delivery Reviews·Carsten EhrhardtUdo Bakowsky
Jan 5, 2011·Journal of Cardiovascular Pharmacology and Therapeutics·Komei TanakaYoshifusa Aizawa

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