Extracellular vesicles of U937 macrophage cell line infected with DENV-2 induce activation in endothelial cells EA.hy926

Abstract

Endothelial activation and alteration during dengue virus (DENV) infection are multifactorial events; however, the role of extracellular vesicles (EVs) in these phenomena is not known. In the present study, we characterized the EVs released by DENV-2 infected U937 macrophage cell line and evaluated the changes in the physiology and integrity of the EA.hy926 endothelial cells exposed to them. U937 macrophages were infected, supernatants were collected, and EVs were purified and characterized. Then, polarized endothelial EA.hy926 cells were exposed to the EVs for 24 h, and the transendothelial electrical resistance (TEER), monolayer permeability, and the expression of tight junction and adhesion proteins and cytokines were evaluated. The isolated EVs from infected macrophages corresponded to exosomes and apoptotic bodies, which contained the viral NS3 protein and different miRs, among other products. Exposure of EA.hy926 cells to EVs induced an increase in TEER, as well as changes in the expression of VE-cadherin and ICAM in addition leads to an increase in TNF-α, IP-10, IL-10, RANTES, and MCP-1 secretion. These results suggest that the EVs of infected macrophages transport proteins and miR that induce early changes in the physiology of the endothelium, leading to its activation and eliciting a defense program against damage during first stages of the disease, even in the absence of the virus. OPEN ACCESS Citation: Velandia-Romero ML, Calderón-Peláez MA, Balbás-Tepedino A, Márquez-Ortiz RA, Madroñero LJ, Barreto Prieto A, et al. (2020) Extracellular vesicles of U937 macrophage cell line infected with DENV-2 induce activation in endothelial cells EA.hy926. PLoS ONE 15(1): e0227030. https://doi.org/10.

Macrophage-derived extracellular vesicles: diverse mediators of pathology and therapeutics in multiple diseases

“…During the early stage of fungal infection, EV contents from Candida albicans -infected macrophages were extensively altered and were intimately associated with enhanced cytokine secretion in macrophages 121 . Moreover, EVs from dengue virus (DENV)-infected macrophages also transfer the non-structural protein (NS3) encoded by DENV RNA to promote cytokine release in ECs, activating the defense program against dengue virus infection at the early stage 122 . EVs released from toll-like receptor 3 (TLR3)-activated macrophages, which are often blocked during infection, were found to possess abundant miR-29 to inhibit HCV replication in hepatocytes 123 .…”

SARS-CoV-2 Infection Reaches the Human Nervous System: How?

“…The isolated EVs released from DENV-2 infected U937 macrophage cell line carrying the viral NS3 protein and different miRs induced an increase in the polarization of the endothelial (EA.hy926) monolayer cells permeability, as well as changes in the expression of ICAM and the VE-cadherin , also leading to an increase in the levels of the IP-10, TNF-α, RANTES, IL-10, and MCP-1 secretion, even in the absence of the virus [202], suggesting that a proinflammatory status was involved in the endothelial permeability alteration. The miRs most frequently counted within the vesicles obtained from such DENV infected cells include the miR 21, miR 92a, and miR 191, which are strongly associated with many biological pathways involving endothelial cell processes, such as tubular network formation, angiogenesis, and brain microvascular reparation.…”

“…The miRs most frequently counted within the vesicles obtained from such DENV infected cells include the miR 21, miR 92a, and miR 191, which are strongly associated with many biological pathways involving endothelial cell processes, such as tubular network formation, angiogenesis, and brain microvascular reparation. Such vesicles obtained from the DENV infected cells induced an endothelial activation, possibly determined by the miR that they contain [202].…”

Extracellular Vesicles in Viral Spread and Antiviral Response

“…Host EVs may enter mosquito cells, inhibiting their infection through the restriction of virus-endosomal membrane fusion [ 83 ]. In addition, exosomes released from DENV-infected macrophages and added to the human EA.hy926 endothelial cells induced physiological changes in that cell line, leading to a protective effect during the early stages of infection that may help to maintain endothelial integrity [ 84 ]. Exosomes from C6/36 mosquito cells infected with Zika virus (ZIKV) may also modify host cells responses and contribute to the pathogenesis of ZIKV infection in humans.…”