Altered Moesin and Actin Cytoskeleton Protein Rearrangements Affect Transendothelial Permeability in Human Endothelial Cells upon Dengue Virus Infection and TNF-α Treatment

• Published in: Viruses Vol. 13; no. 10; p. 2042
• Main Authors: Suttitheptumrong, Aroonroong, Mahutchariyakul, Thanaporn, Rawarak, Nantapon, Reamtong, Onrapak, Boonnak, Kobporn, Pattanakitsakul, Sa-Nga
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 11.10.2021; MDPI
• Subjects: Actin; Actin Cytoskeleton - metabolism; Actins - metabolism; Capillary Permeability; Chemokines; Cloning; Collagen; Cytokines; Cytoskeleton; Cytoskeleton - metabolism; cytoskeleton protein; Dengue - metabolism; Dengue fever; Dengue hemorrhagic fever; Dengue Virus - metabolism; dengue virus infection; Electrical resistivity; Endothelial cells; Endothelial Cells - virology; Flow cytometry; Humans; Immunofluorescence; Infections; Mass spectroscopy; Microfilament Proteins - metabolism; Moesin; Monoclonal antibodies; Permeability; Proteins; Proteomics; TNF-alpha; transendothelial permeability; Tumor Necrosis Factor-alpha - metabolism; Tumor necrosis factor-TNF; Tumor necrosis factor-α; Vinculin; Viral infections; United States > US; transendothelial permeability; dengue virus infection; TNF-alpha; cytoskeleton protein
• ISSN: 1999-4915; 1999-4915
• DOI: 10.3390/v13102042

It has been hypothesized that the host, viral factors, and secreted cytokines (especially TNF-α) play roles in the pathogenesis of secondary dengue infections. Mass spectrometry-based proteomic screening of cytoskeleton fractions isolated from human endothelial (EA.hy926) cells upon dengue virus (DENV) infection and TNF-α treatment identified 450 differentially altered proteins. Among them, decreased levels of moesin, actin stress fiber rearrangements, and dot-like formations of vinculin were observed with western blot analyses and/or immunofluorescence staining (IFA). In vitro vascular permeability assays using EA.hy926 cells, seeded on collagen-coated transwell inserts, showed low levels of transendothelial electrical resistance in treated cells. The synergistic effects of DENV infection and TNF-α treatment caused cellular permeability changes in EA.hy926 cells, which coincided with decreasing moesin levels and the production of abnormal organizations of actin stress fibers and vinculin. Functional studies demonstrated moesin overexpression restored transendothelial permeability in DENV/TNF-α-treated EA.hy926 cells. The present study improves the understanding of the disruption mechanisms of cytoskeleton proteins in enhancing vascular permeability during DENV infection and TNF-α treatment. The study also suggests that these disruption mechanisms are major factors contributing to vascular leakage in severe dengue patients.