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In Vitro SARS-CoV-2 Infection of Microvascular Endothelial Cells: Effect on Pro-Inflammatory Cytokine and Chemokine Release
Authors:Maria Dolci  Lucia Signorini  Sarah D&#x;Alessandro  Federica Perego  Silvia Parapini  Michele Sommariva  Donatella Taramelli  Pasquale Ferrante  Nicoletta Basilico  Serena Delbue
Affiliation:1.Department of Biomedical, Surgical and Dental Sciences, University of Milan, Via Pascal 36, 20133 Milan, Italy; (M.D.); (L.S.); (F.P.); (P.F.); (S.D.);2.Department of Pharmacological and Biomolecular Sciences, University of Milan, Via Pascal 36, 20133 Milan, Italy; (S.D.); (D.T.);3.Department of Biomedical Sciences for Health, University of Milan, Via Pascal 36, 20133 Milan, Italy;4.Department of Biomedical Sciences for Health, University of Milan, Milan, Mangiagalli 31, 20133 Milan, Italy;
Abstract:In the novel pandemic of Coronavirus Disease 2019, high levels of pro-inflammatory cytokines lead to endothelial activation and dysfunction, promoting a pro-coagulative state, thrombotic events, and microvasculature injuries. The aim of the present work was to investigate the effect of SARS-CoV-2 on pro-inflammatory cytokines, tissue factor, and chemokine release, with Human Microvascular Endothelial Cells (HMEC-1). ACE2 receptor expression was evaluated by western blot analysis. SARS-CoV-2 infection was assessed by one-step RT-PCR until 7 days post-infection (p.i.), and by Transmission Electron Microscopy (TEM). IL-6, TNF-α, IL-8, IFN-α, and hTF mRNA expression levels were detected by RT-PCR, while cytokine release was evaluated by ELISA. HMEC-1 expressed ACE2 receptor and SARS-CoV-2 infection showed a constant viral load. TEM analysis showed virions localized in the cytoplasm. Expression of IL-6 at 24 h and IFN-α mRNA at 24 h and 48 h p.i. was higher in infected than uninfected HMEC-1 (p < 0.05). IL-6 levels were significantly higher in supernatants from infected HMEC-1 (p < 0.001) at 24 h, 48 h, and 72 h p.i., while IL-8 levels were significantly lower at 24 h p.i. (p < 0.001). These data indicate that in vitro microvascular endothelial cells are susceptible to SARS-CoV-2 infection but slightly contribute to viral amplification. However, SARS-CoV-2 infection might trigger the increase of pro-inflammatory mediators.
Keywords:Human Microvascular Endothelial Cells (HMEC-1)  SARS-CoV-2  inflammatory mediators
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