Background: Inflammatory cytokines dysregulate microvascular function, yet how cytokines affect lymphatic endothelial cells (LEC) are unclear. Methods and Results: We examined effects of TNF-a, IL-1b, and IFN-g on LEC proliferation, endothelial cell adhesion molecule (ECAM) expression, capillary formation, and barrier changes in murine (SV-LEC) and human LECs (HMEC-1a). Results: All cytokines induced ICAM-1, VCAM-1, MAdCAM-1, and E-selectin in SV-LECs; TNF-a, IL-1b and IFN-g induced ECAMs (but not MAdCAM-1) in HMEC-1a. IL-1b increased, while IFN-g and TNF-a reduced SV-LEC proliferation. While TNF-a induced, IFN-g decreased, and IL-1b did not show any effect on HMEC-1a proliferation. TNF-a, IL-1b, and IFN-g each reduced capillary formation in SV-LEC and in HMEC-1a. TNF-a and IL-1b reduced barrier in SV-LEC and HMEC-1a; IFN-g did not affect SV-LEC barrier, but enhanced HMEC-1a barrier. Inflammatory cytokines alter LEC growth, activation and barrier function in vitro and may disturb lymphatic clearance increasing tissue edema in vivo. Conclusion: Therapies that maintain or restore lymphatic function (including cytokines blockade), may represent important strategies for limiting inflammation.