Dengue hemorrhagic fever is characterized by a unique vascular leakage syndrome. The mechanisms of endothelial barrier dysfunction in dengue hemorrhagic fever are not well understood. We examined the modulation of endothelial barrier function in dengue virus type 2 (DENV2) infections using primary human umbilical vein endothelial cells. We demonstrated that the increase in endothelial barrier function within 72 hours after DENV2 infection is mediated by type I interferon-dependent CD73 up-regulation. After 72 hours, DENV2 slowed the recovery of endothelial barrier function in response to tumor necrosis factor-alpha or vascular endothelial growth factor. This phenomenon was likely caused by type I interferon receptor signaling inhibition and lower CD73 levels in DENV2-infected endothelial cells. Our findings suggest that during DENV2 infection, endothelial barrier homeostasis is maintained by a balance between pro-inflammatory and pro-angiogenic cytokines, and type I interferon-dependent CD73 expression and activity.

Dengue virus (DENV), a re-emerging arbovirus, readily infects dendritic cells (DC) in culture and in vivo. However, there have been contradictory reports regarding the effect of DENV infection on DC activation and maturation. DC undergo a series of functional changes following exposure to infectious agents, including cytokine production and costimulatory and MHC molecule induction, culminating in stimulation of adaptive immune responses. Immunological memory to primary DENV infection critically influences disease severity during subsequent infections with heterologous serotypes. To explore these phenomena, we examined DENV infection-dependent and -independent effects on DC secretory, phenotypic, and allostimulatory functions. DENV infection of DC resulted in the secretion of a broad array of cytokines and chemokines. Type I IFN produced by DC inhibited propagation of infection and induced the chemokine IFN-gamma-inducible protein 10 (IP-10; CXCL10). Based on intracellular cytokine staining, infected DC produced less IP-10 but more TNF-alpha than uninfected bystander cells in the same culture. DENV exposure activated surface molecule expression on infected and bystander cells; infected DC had enhanced programmed death ligand 2 (PD-L2) and MHC II expression but reduced levels of PD-L1, CD80, CD86, and MHC I relative to bystander DC. Dengue-infected DC cultures stimulated resting allogeneic CD4 T cell proliferation, although an increasing multiplicity of infection was associated with decreasing stimulatory capacity of DC. These data demonstrate that functional maturation of DC in response to dengue infection is modified by the presence of virus through IFN-dependent and -independent mechanisms with consequences for the development of adaptive immunity.