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Related Experiment Videos

Glycolipids support E-selectin-specific strong cell tethering under flow.

M M Burdick1, B S Bochner, B E Collins

  • 1Department of Chemical Engineering, Johns Hopkins University, Baltimore, Maryland 21218, USA.

Biochemical and Biophysical Research Communications
|May 26, 2001
PubMed
Summary
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Glycosphingolipids with specific structures act as ligands for E-selectin, mediating cell rolling. However, they do not bind to P-selectin, indicating selective interactions in cell adhesion.

Area of Science:

  • Cellular Biology
  • Biochemistry
  • Immunology

Background:

  • Selectins are cell adhesion molecules crucial for immune cell trafficking.
  • Glycosphingolipids are complex lipids involved in cell surface recognition.
  • Understanding selectin-ligand interactions is vital for studying inflammatory diseases.

Purpose of the Study:

  • To investigate the role of glycosphingolipids as ligands for E-selectin and P-selectin.
  • To determine the functional differences in cell adhesion mediated by these selectins.

Main Methods:

  • Perfusion of Chinese hamster ovary (CHO) cells expressing E-selectin (CHO-E) or P-selectin (CHO-P) over immobilized alpha2,3-sialyl Lewis X (alpha2,3-sLe(x)) glycosphingolipids.
  • Analysis of cell tethering and rolling interactions under varying shear stress, selectin, and ligand densities.

Related Experiment Videos

  • Application of a stochastic model to quantify cell rolling parameters.
  • Detachment assays to assess adhesive strengths.

    • Main Results:

    • CHO-E cells exhibited extensive tethering and stable rolling on alpha2,3-sLe(x) glycosphingolipids, but not on alpha2,6-sLe(x).
    • CHO-P cells showed limited, fast rolling on alpha2,3-sLe(x) glycosphingolipids.
    • Stochastic modeling revealed differences in bond release dynamics between CHO-E and CHO-P cells.
    • Detachment assays confirmed stronger adhesion of CHO-E cells to the glycosphingolipid ligand.

    Conclusions:

    • Glycosphingolipids expressing alpha2,3-sialyl Lewis X are functional ligands for E-selectin, mediating significant cell tethering and rolling.
    • P-selectin interacts weakly with these glycosphingolipids, supporting only limited cell rolling.
    • These findings highlight the selectivity of E-selectin and P-selectin binding to specific glycosphingolipid structures.