A method for measuring leukocyte rolling on the selectins.
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.
Schmuke
J J
JJ
Department of Immunology and Glycobiology, G.D. Searle/Monsanto Company, St. Louis, Missouri 63167, USA.
Welply
J K
JK
D015815:Cell Adhesion Molecules / Q000378:metabolism; D002465:Cell Movement; D016264:Dextran Sulfate / Q000378:metabolism; D019040:E-Selectin; D004492:Edetic Acid / Q000494:pharmacology; D006801:Humans; D007074:Immunoglobulin G; D007962:Leukocytes / Q000502:physiology*; D008856:Microscopy, Fluorescence; D009844:Oligosaccharides / Q000378:metabolism / Q000494:pharmacology; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D011134:Polysaccharides / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D000080506:Sialyl Lewis X Antigen; D014407:Tumor Cells, Cultured
D015815:Cell Adhesion Molecules; D019040:E-Selectin; D007074:Immunoglobulin G; D009844:Oligosaccharides; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins; D011134:Polysaccharides; D011993:Recombinant Fusion Proteins; D000080506:Sialyl Lewis X Antigen; D016264:Dextran Sulfate; C007789:fucoidan; D004492:Edetic Acid