Neutrophil recruitment limited by high-affinity bent β2 integrin binding ligand in cis

Neutrophils are essential for innate immunity and inflammation and many neutrophil functions are beta(2) integrin-dependent. Integrins can extend (E+) and acquire a high-affinity conformation with an 'open' headpiece (H+). The canonical switchblade model of integrin activation proposes that the E+ conformation precedes H+, and the two are believed to be structurally linked. Here we show, using high-resolution quantitative dynamic footprinting (qDF) microscopy combined with a homogenous conformation-reporter binding assay in a microfluidic device, that a substantial fraction of beta(2) integrins on human neutrophils acquire an unexpected E-H+ conformation. E-H+ beta(2) integrins bind intercellular adhesion molecules (ICAMs) in cis, which inhibits leukocyte adhesion in vitro and in vivo. This endogenous anti-inflammatory mechanism inhibits neutrophil aggregation, accumulation and inflammation.