A novel multi-parameter assay to dissect the pharmacological effects of different modes of integrin αLβ2 inhibition in whole blood

Br J Pharmacol. 2015 Oct;172(20):4875-87. doi: 10.1111/bph.13256. Epub 2015 Oct 19.


Background and purpose: The integrin αLβ2 plays central roles in leukocyte adhesion and T cell activation, rendering αLβ2 an attractive therapeutic target. Compounds with different modes of αLβ2 inhibition are in development, currently. Consequently, there is a foreseeable need for bedside assays, which allow assessment of the different effects of diverse types of αLβ2 inhibitors in the peripheral blood of treated patients.

Experimental approach: Here, we describe a flow cytometry-based technology that simultaneously quantitates αLβ2 conformational change upon inhibitor binding, αLβ2 expression and T cell activation at the single-cell level in human blood. Two classes of allosteric low MW inhibitors, designated α I and α/β I allosteric αLβ2 inhibitors, were investigated. The first application revealed intriguing inhibitor class-specific profiles.

Key results: Half-maximal inhibition of T cell activation was associated with 80% epitope loss induced by α I allosteric inhibitors and with 40% epitope gain induced by α/β I allosteric inhibitors. This differential establishes that inhibitor-induced αLβ2 epitope changes do not directly predict the effect on T cell activation. Moreover, we show here for the first time that α/β I allosteric inhibitors, in contrast to α I allosteric inhibitors, provoked partial downmodulation of αLβ2, revealing a novel property of this inhibitor class.

Conclusions and implications: The multi-parameter whole blood αLβ2 assay described here may enable therapeutic monitoring of αLβ2 inhibitors in patients' blood. The assay dissects differential effect profiles of different classes of αLβ2 inhibitors.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biological Assay
  • Epitopes / immunology
  • Flow Cytometry
  • Humans
  • Lymphocyte Activation
  • Lymphocyte Function-Associated Antigen-1 / blood*
  • Naphthalenes / pharmacology*
  • Oxazines / pharmacology*
  • Phthalic Acids / pharmacology*
  • T-Lymphocytes / immunology
  • beta-Alanine / analogs & derivatives*
  • beta-Alanine / pharmacology


  • Epitopes
  • LFA 878
  • Lymphocyte Function-Associated Antigen-1
  • Naphthalenes
  • Oxazines
  • Phthalic Acids
  • XVA 143
  • beta-Alanine