JAKinibs prevent persistent, IFNγ-autonomous endothelial cell inflammation and immunogenicity.

The adhesion and subsequent activation of T cells is a critical step in local inflammatory responses, particularly of alloreactive leukocytes in rejection of transplanted donor tissue. Interferon (IFN)γ is an adaptive cytokine that promotes endothelial cell (EC) expression of pro-adhesive factors and costimulatory molecules. We recently reported that IFNγ-induced endothelial cell antigen-presenting capacity was protracted after cytokine withdrawal. This study sought to determine what intracellular signaling mediates this chronic endothelial activation by IFNγ. The durability of interferon signaling in human aortic endothelial activation was tested. Pro-adhesive and costimulatory gene expression, phenotype, secretome, and Janus kinase (JAK)/STAT phosphorylation in human primary endothelial cells were measured under chronic and transient IFNγ stimulation, with various JAK inhibitors. IFNγ reporter cells were tested for STAT1 transcriptional activity with JAK inhibition and suppressors of cytokine signaling (SOCS) overexpression, under continuous and priming conditions. The consequences of even short exposure to IFNγ were long-lasting and broad, with sustained elevation of adhesion molecules and chemokines up to 48 h later. JAK/STAT and interferon response factor expression were likewise durable, dependent on new transcription but autonomous of continuous IFNγ. Persistent STAT new transcription and JAK signaling in the endothelium was required to maintain a pro-adhesive and proimmunogenic phenotype after IFNγ withdrawal since both could be prevented by cycloheximide but only by JAKinibs with potency against JAK2. Finally, the suppressor of cytokine signaling SOCS1 failed to emerge in primed endothelial cells, which likely accounted for prolonged inflammatory gene expression. The results reveal a sustained JAK-dependent perturbation of endothelial function and suggest that JAKinibs may have therapeutic benefits in dampening vascular inflammation and allogeneic leukocyte activation.