Endothelial cell activation driven by CD4+ memory T cells produced soluble factors

In this project we analyzed whole transcriptome changes in endothelial cells (ECs) in response to stimulation with activated CD4+ memory T cells produced factors. Given the central role of NF-κB signaling in EC activation we treated cells with inhibitors for canonical NF-κB signaling (IKKβ inhibitor, iIKKβ) and non-canonical NF-κB signaling (NF-κB inducting kinase (NIK) inhibitor, iNIK). Cells were stimulated 72h with conditioned medium (hereafter called Tm sup = 50% culture supernatant from aCD3/aCD28 activated CD4+ memory T cells; 50% RMPI supplemented with 10% FCS) and treated with iIKKβ, iNIK or DMSO as a control. Other controls taken along were unstimulated DMSO treated ECs and unstimulated untreated ECs. Examination of whole transcriptome changes in primary human umbilical cord ECs (HUVEC) stimulated with Tm sup and treated with iIKKβ or iNIK.