title: The cytokine response of U937-derived macrophages infected through antibody-dependent enhancement of dengue virus disrupts cell apical-junction complexes and increases vascular permeability.
abstract: 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.
authors.affiliation: Department of Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies (CINVESTAV), Mexico City, DF, Mexico.
authors.forename: Henry
authors.initials: H
authors.lastname: Puerta-Guardo
authors.forename: Arturo
authors.initials: A
authors.lastname: Raya-Sandino
authors.forename: Lorenza
authors.initials: L
authors.lastname: González-Mariscal
authors.forename: Victor H
authors.initials: VH
authors.lastname: Rosales
authors.forename: José
authors.initials: J
authors.lastname: Ayala-Dávila
authors.forename: Bibiana
authors.initials: B
authors.lastname: Chávez-Mungía
authors.forename: Daniel
authors.initials: D
authors.lastname: Martínez-Fong
authors.forename: Fernando
authors.initials: F
authors.lastname: Medina
authors.forename: Juan E
authors.initials: JE
authors.lastname: Ludert
authors.forename: Rosa María
authors.initials: RM
authors.lastname: del Angel
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D019067:Antibody-Dependent Enhancement / Q000276:immunology*; D015153:Blotting, Western; D002199:Capillary Permeability / Q000187:drug effects / Q000276:immunology*; D002470:Cell Survival / Q000502:physiology; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000494:pharmacology; D016207:Cytokines / Q000276:immunology* / Q000378:metabolism; D003715:Dengue / Q000276:immunology / Q000503:physiopathology*; D003716:Dengue Virus / Q000276:immunology*; D004285:Dogs; D017097:Electric Impedance; D005070:Evans Blue; D005455:Fluorescent Antibody Technique; D006801:Humans; D008264:Macrophages / Q000378:metabolism / Q000648:ultrastructure / Q000821:virology*; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D046529:Microscopy, Electron, Transmission; D019108:Tight Junctions / Q000378:metabolism; D020298:U937 Cells
chemical_list: D017077:Culture Media, Conditioned; D016207:Cytokines; D005070:Evans Blue