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A method for measuring leukocyte rolling on the selectins

J J Schmuke1, J K Welply

  • 1Department of Immunology and Glycobiology, G.D. Searle/Monsanto Company, St. Louis, Missouri 63167, USA.

Analytical Biochemistry
|April 10, 1995
PubMed
Summary
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A new method simulates leukocyte rolling in microvasculature using capillary tubes coated with selectins. This high-throughput technique enables efficient testing of potential antagonists for cell rolling inhibition.

Area of Science:

  • Biomedical Engineering
  • Cell Biology
  • Immunology

Background:

  • Leukocyte rolling is a critical step in immune cell trafficking within the microvasculature.
  • Understanding leukocyte-endothelial cell interactions is vital for developing anti-inflammatory therapies.

Purpose of the Study:

  • To develop an inexpensive, high-throughput method for simulating leukocyte rolling in vitro.
  • To assess the utility of this method for screening potential antagonists of leukocyte rolling.

Main Methods:

  • Utilized fused silica capillary tubes coated with E- or P-selectin.
  • Introduced fluorescently labeled HL-60 cells at a constant flow rate.
  • Monitored cell rolling using fluorescence microscopy and recorded images via time-lapse video.

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Main Results:

  • Rolling incidence and velocity correlated with selectin concentration on the capillary tube.
  • sLex(Glc) tetrasaccharide inhibited HL-60 cell rolling on E-selectin.
  • Fucoidan and dextran sulfate inhibited HL-60 cell rolling on P-selectin.

Conclusions:

  • The developed capillary tube assay is an effective, high-throughput method for studying leukocyte rolling.
  • The assay's small volume requirement makes it suitable for testing limited quantities of potential antagonists.
  • This technique facilitates the identification of compounds that modulate selectin-mediated leukocyte adhesion.