TNF-a induces VE-cadherin-dependent gap/JAIL cycling through an intermediate state essential for neutrophil transmigration

Abstract

Inflammatory endothelial phenotypes describe distinct cellular patterns essential for controlling transendothelial migration of leukocytes (TEM). While TNF--induced CAM expression mediates leukocyte interaction, the role of a potential inflammatory morphological phenotype (IMP) - characterised by barrier-function decrease and shape-change in TEM - remains unclear. This study identifies the TNF--induced IMP as indispensable for neutrophil TEM, while regulating barrier-function. The TNF--induced IMP progresses through two states: an intermediate state that transiently enhances barrier function via MLC-dephosphorylation, junctional actin recruitment and VE-cadherin linearisation, protecting the monolayer from collapse; while the subsequent development of the IMP requires MLC rephosphorylation, junctional actin disassembly, stress fibre formation and Arp2/3-mediated membrane protrusions causing shape-change. This in turn dilutes junctional VE-cadherin, forming intercellular gaps for neutrophil TEM, while inducing junction-associated intermittent lamellipodia (JAIL) to locally restore VE-cadherin adhesion, appearing as gap/JAIL cycles driving junctional dynamics. VE-cadherin overexpression blocks TNF--induced IMP and gap/JAIL cycling, reducing TEM by [~]80% without altering CAM expression. These findings highlight gap/JAIL cycling and MLC phosphorylation as key IMP regulators and potential therapeutic targets for inflammatory diseases.