Severe dengue (SD) is a life-threatening complication of
dengue that includes vascular permeability syndrome (VPS) and respiratory distress.
Secondary infections are considered a risk factor for developing SD, presumably through a mechanism called antibody-dependent enhancement (ADE). Despite extensive studies, the molecular bases of how ADE contributes to SD and VPS are largely unknown. This work compares the
cytokine responses of differentiated U937 human monocytic cells infected directly with dengue virus (DENV) or in the presence of enhancing concentrations of a humanized
monoclonal antibody recognizing
protein E (ADE-DENV
infection). Using a cytometric bead assay, ADE-DENV-infected cells were found to produce significantly higher levels of the proinflammatory
cytokines interleukin 6 (IL-6), IL-12p70, and
tumor necrosis factor alpha (TNF-α), as well as
prostaglandin E2 (
PGE2), than cells directly infected. The capacity of conditioned supernatants (
conditioned medium [CM]) to disrupt tight junctions (TJs) in MDCK cell cultures was evaluated. Exposure of MDCK cell monolayers to CM collected from ADE-DENV-infected cells (ADE-CM) but not from cells infected directly led to a rapid loss of transepithelial electrical resistance (TER) and to delocalization and degradation of apical-junction complex
proteins. Depletion of either TNF-α,
IL-6, or IL-12p70 from CM from ADE-DENV-infected cells fully reverted the disrupting effect on TJs. Remarkably, mice injected intraperitoneally with ADE-CM showed increased vascular permeability in sera and lungs, as indicated by an
Evans blue quantification assay. These results indicate that the
cytokine response of U937-derived macrophages to ADE-DENV
infection shows an increased capacity to disturb TJs, while results obtained with the mouse model suggest that such a response may be related to the vascular plasma leakage characteristic of SD.