The central nervous system (CNS) is an immune privileged site where highly specialised endothelial cells, which comprise the blood-brain barrier (BBB), acts as a selectively permeable interface to control the passage of nutrients and inflammatory cells into the brain.  Disruption of the BBB and leukocyte infiltration are among the abnormalities seen in neuroinflammatory diseases such as multiple sclerosis. Using a recently developed human cerebral microvascular endothelial cell line (hCMVEC), we investigated the effects of two pro-inflammatory cytokines IL-1β and TNFα on the inflammatory response of these cells.  Specifically, temporal expression of cell adhesion molecules ICAM-1 (CD54) and VCAM-1 (CD106), and cytokine secretion were investigated using flow cytometry and multiplex cytokine arrays. The endothelial cells demonstrated differential expression of cell adhesion molecules, where TNF-α induced higher expression of ICAM-1 and VCAM-1 over the time course of 6 days. Quantification of soluble ICAM-1 and soluble VCAM-1 in endothelial conditioned media revealed that more adhesion molecules were cleaved from the cell surface when cells were stimulated by IL-1β as opposed to TNFα over a period of 72 hours. IL-1β induced a substantially different cytokine secretion profile in comparison to TNFα.  Our study shows that these two key pro-inflammatory cytokines differentially regulate the inflammatory response of brain endothelial cells.