Motile Human Neutrophils Sense Ligand Density Over Their Entire Contact Area

Steven J. Henry, John C. Crocker, Daniel A. Hammer

Neutrophils are key components of the immune system and motility is central their function during the inflammatory response. We have previously demonstrated that neutrophils are capable of switching their motile phenotype between amoeboid-like and keratocyte-like in response to the ligand density of adhesion molecules (Henry et al. in Int Biol 6:348-356, 2014). In this study, we engineered planar micropatterned surfaces that presented adhesion molecules in local islands of high density, separated by regions largely devoid of ligands. By controlling the geometry of islands we made arrays in which the local (on island) adhesion density was high but the global (multi-island) adhesion density over the entire cell-substrate interface was low. Neutrophils in contact with these island arrays assumed a well-spread and directionally-persistent motile phenotype (keratocyte-like) in contrast to the classical amoeboid morphology they display on uniform fields of high adhesion density. By virtue of our rationally designed substrates, we were able to conclude that neutrophils were integrating the stimulation received across their entire contact interface; furthermore, they were able to mount this whole cell response on the timescale of seconds. This work demonstrates the capacity of adhesive microenvironments to direct the phenotype of cell motility, which has broader implications in physiologic processes such as inflammation and cancer metastasis.