Immunology & Immunological Disorders Poster Session



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Effect of Anticoagulant on the Measurement of Neutrophil L-selectin Levels

Fagan, D. (Department of Biological Sciences, Youngstown State University, USA)
Smith, T. (Department of Biological Sciences, USA)
Niehaus, G. (Department of Physiology, Neucom, USA)

Contact Person: Diana L Fagan (dlfagan@cc.ysu.edu)


L-selectin (CD62L) is a cell adhesion molecule that helps to direct neutrophils to the site of an inflammatory response. Stimulation of white blood cells and subsequent migratrion of the cells into the tissues results in proteolytic cleavage of L-selectin from the cell surface. Four anticoagulants (EDTA, potassium oxalate, sodium citrate and heparin) were compared for their effect on neutrophil L-selectin levels. Blood was drawn from seven volunteers into tubes containing one of the anticoagulants, incubated at room temperature for various time points (up to 1 hr), then placed on ice. Cells were labeled with fluorescent antibodies directed against L-selectin and fluorescence measured using flow cytometry. When the cells were collected in EDTA, no significant variation in neutrophil L-selectin levels was seen at any time point. L-selectin levels decreased 40% when the cells were collected in heparin or sodium citrate. This effect was not statistically significant, but reflected the large degree of variability seen between samples in the presence of these anticoagulants. A significant decrease in L-selectin (60%) was seen in the presence of potassium oxalate (p = 0.00096). The results demonstrate that collection of blood in EDTA resulted in the least sample variability and did not contribute to the shedding of L-selectin.

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Poster Number PAfagan0456
Keywords: L-selectin, CD62L, anticoagulant, adhesion

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Fagan, D.; Smith, T.; Niehaus, G.; (1998). Effect of Anticoagulant on the Measurement of Neutrophil L-selectin Levels. Presented at INABIS '98 - 5th Internet World Congress on Biomedical Sciences at McMaster University, Canada, Dec 7-16th. Available at URL http://www.mcmaster.ca/inabis98/immunology/fagan0456/index.html
© 1998 Author(s) Hold Copyright