Zika Virus Infection Induces Elevation of Tissue Factor Production and Apoptosis on Human Umbilical Vein Endothelial Cells

Abstract

Zika virus (ZIKV) infection is typically characterized by a mild disease presenting with fever, maculopapular rash, headache, fatigue, myalgia, and arthralgia. A recent animal study found that ZIKV-infected pregnant Ifnar −/− mice developed vascular damage in the placenta and reduced amount of fetal capillaries. Moreover, ZIKV infection causes segmental thrombosis in the umbilical cord of pregnant rhesus macaques. Furthermore, several case reports suggest that ZIKV infection cause coagulation disorders. These results suggest that ZIKV could cause an alteration in the host hemostatic response, however, the mechanism has not been investigated thus far. This paper aims to determine whether ZIKV infection on HUVECs induces apoptosis and elevation of tissue factor (TF) that leads to activation of secondary hemostasis. We infected HUVECs with two ZIKV strains and performed virus titration, immunostaining, and flow cytometry to confirm and quantify infection. We measured TF concentrations with flow cytometry and performed thrombin generation test (TGT) as a functional assay to assess secondary hemostasis. Furthermore, we determined the amount of cell death using flow cytometry. We also performed enzyme-linked immunosorbent assay (ELISA) to determine interleukin (IL)-6 and IL-8 production and conducted blocking experiments to associate these cytokines with TF expression. Both ZIKV strains infected and replicated to high titers in HUVECs. We found that infection induced elevation of TF expressions. We also showed that increased TF expression led to shortened TGT time. Moreover, the data revealed that infection induced apoptosis. In addition, there was a significant increase of IL-6 and IL-8 production in infected cells. Here we provide in vitro evidence that infection of HUVECs with ZIKV induces apoptosis and elevation of TF expression that leads to activation of secondary hemostasis.

RIG-I Plays a Dominant Role in the Induction of Transcriptional Changes in Zika Virus-Infected Cells, which Protect from Virus-Induced Cell Death

“…ZIKV infection causes apoptosis [ 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 ]. We therefore asked whether reduced innate immune recognition of ZIKV in RIG-I KO cells impacts virus-induced cell death.…”

Participation of Extracellular Vesicles from Zika-Virus-Infected Mosquito Cells in the Modification of Naïve Cells’ Behavior by Mediating Cell-to-Cell Transmission of Viral Elements

“…Dengue virus has also been shown to cause coagulation disorders in ECs [54]. Anfasa et al (2019) provided in vitro evidence that ZIKV infection of human umbilical vein endothelial cells (HUVECs) induces apoptosis and increases TF production, which triggers the activation of secondary hemostasis [68]. Additionally, to evaluate the possible expression of a pro-inflammatory response by EVs released from ZIKV-infected C6/36 cells, we measured the TNF-α mRNA expression in naïve ECs infected by ZIKV (MOI 1) and those stimulated with the mock C6/36 EVs or the ZIKV C6/36 EVs (small/large isolates the same for RNase A + UV-treated).…”

Endocytosis of flavivirus NS1 is required for NS1-mediated endothelial hyperpermeability and is abolished by a single N-glycosylation site mutation

“…Regarding overall pathogenic mechanisms, our investigation specifically focuses on a cell-intrinsic pathway by which NS1 mediates the disruption of the EGL, but flavivirus-induced vascular dysfunction is a complex and multifactorial process. It can be mediated through the NS1 endothelial cell-intrinsic pathway [9, 10, 36, 37], through production of proinflammatory cytokines by immune cells induced by either the virus or NS1 [11, 47], through NS1-activated platelets [48], or even potentially through direct viral infection leading to apoptosis of endothelial cells [49, 50]. Though endothelial cells can be infected in vitro by DENV, findings from autopsy studies suggest that this does not occur in vivo [22, 51, 52].…”