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Related Experiment Videos

CD11b/CD18-coated microspheres attach to E-selectin under flow.

K L Crutchfield1, V R Shinde Patil, C J Campbell

  • 1Department of Biomedical Engineering, The University of Memphis, Tennessee, USA.

Journal of Leukocyte Biology
|February 12, 2000
PubMed
Summary
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Leukocyte function-associated antigen 1 (LFA-1) and Mac-1 (also known as alphaMbeta2) support neutrophil adhesion to E-selectin. This study demonstrates that alphaMbeta2 directly mediates attachment to E-selectin under flow conditions.

Area of Science:

  • Immunology
  • Cell Biology
  • Biophysics

Background:

  • Neutrophils adhere to E-selectin under physiological flow conditions.
  • Sialyl Lewis X (SLe(x))-type glycans are proposed ligands for E-selectin.
  • Leukocyte beta2 integrins are glycosylated with SLe(x), suggesting a potential role in E-selectin binding.

Purpose of the Study:

  • To investigate whether beta2 integrins can mediate attachment to E-selectin.
  • To test the hypothesis that alphaMbeta2 supports adhesion to E-selectin.

Main Methods:

  • Coating 10-microm microspheres with purified CD11b/CD18 (alphaMbeta2).
  • Investigating the adhesion of alphaMbeta2-coated microspheres to E-selectin expressing cells (CHO-E) and activated human umbilical vein endothelial cells (HUVEC) under in vitro flow conditions.

Related Experiment Videos

  • Utilizing a monoclonal antibody (mAb) against E-selectin to block adhesion events.

    • Main Results:

    • alphaMbeta2 microspheres attached to both CHO-E cells and activated HUVEC under flow.
    • Attachment was eliminated by pretreatment with an anti-E-selectin mAb at a shear stress of 1.8 dynes/cm2.
    • alphaMbeta2 microspheres did not attach to untransfected CHO cells or unactivated HUVEC at the tested shear stress.

    Conclusions:

    • The CD11b/CD18-E-selectin interaction possesses sufficient biophysical properties to mediate attachment.
    • This interaction can support the adhesion of neutrophil-sized particles to E-selectin under flow conditions.
    • Beta2 integrins, specifically alphaMbeta2, play a direct role in mediating neutrophil adhesion to E-selectin.