PT  - JOURNAL ARTICLE
AU  - Nathan H Roy
AU  - Sarah Hyun Ji Kim
AU  - Alexander Buffone, Jr
AU  - Daniel Blumenthal
AU  - Bonnie Huang
AU  - Sangya Agarwal
AU  - Pamela L Schwartzberg
AU  - Daniel A Hammer
AU  - Janis K Burkhardt
TI  - LFA-1 signals to promote actin polymerization and upstream migration in T cells
AID  - 10.1101/2020.04.29.069302
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.04.29.069302
4099  - http://biorxiv.org/content/early/2020/05/02/2020.04.29.069302.short
4100  - http://biorxiv.org/content/early/2020/05/02/2020.04.29.069302.full
AB  - T cell entry into inflamed tissue requires firm adhesion, cell spreading, and migration along and through the endothelial wall. These events require the T cell integrins LFA-1 and VLA-4 and their endothelial ligands ICAM-1 and VCAM-1, respectively. T cells migrate against the direction of shear flow on ICAM-1 and with the direction of shear flow on VCAM-1, suggesting that these two ligands trigger distinct cellular responses. However, the contribution of specific signaling events downstream of LFA-1 and VLA-4 has not been explored. Using primary mouse T cells, we found that engagement of LFA-1, but not VLA-4, induces cell shape changes associated with rapid 2D migration. Moreover, LFA-1 ligation results in activation of the PI3K and ERK pathways, and phosphorylation of multiple kinases and adaptor proteins, while VLA-4 ligation triggers only a subset of these signaling events. Importantly, T cells lacking Crk adaptor proteins, key LFA-1 signaling intermediates, or the ubiquitin ligase cCbl, failed to migrate against the direction of shear flow on ICAM-1. These studies identify novel signaling differences downstream of LFA-1 and VLA-4 that drive T cell migratory behavior.Summary Statement Inflammatory responses require leukocyte migration along the vascular wall. We show that signaling from β2, but not β1, integrins induces cytoskeletal changes needed for upstream migration under shear flow.