Traction forces of neutrophils migrating on compliant substrates

Abstract

Proper functioning of the innate immune response depends on migration of circulating neutrophils into tissues at sites of infection and inflammation. Migration of highly motile, amoeboid cells such as neutrophils has significant physiological relevance, yet the traction forces that drive neutrophil motion in response to chemical cues are not well characterized. To better understand the relationship between chemotactic signals and the organization of forces in motile neutrophils, force measurements were made on hydrogel surfaces under well-defined chemotactic gradients created with a microfluidic device. Two parameters, the mean chemoattractant concentration (C(M)) and the gradient magnitude (Δc/Δx) were varied. Cells experiencing a large gradient with C(M) near the chemotactic receptor K(D) displayed strong punctate centers of uropodial contractile force and strong directional motion on stiff (12 kPa) surfaces. Under conditions of ideal chemotaxis--cells in strong gradients with mean chemoattractant near the receptor K(D) and on stiffer substrates--there is a correlation between the magnitude of force generation and directional motion as measured by the chemotactic index. However, on soft materials or under weaker chemotactic conditions, directional motion is uncorrelated with the magnitude of traction force. Inhibition of either β(2) integrins or Rho-associated kinase, a kinase downstream from RhoA, greatly reduced rearward traction forces and directional motion, although some vestigial lamellipodium-driven motility remained. In summary, neutrophils display a diverse repertoire of methods for organizing their internal machinery to generate directional motion.