Change in the Functional State of Cells of the Human Endothelial Cell Line Ecv-304 Under the Influence of Influenza a Virus, Lipopolysaccharide From Escherichia Coli and Some Drugs

Smirnova, S. S.; Smirnova, Tatiana; Сивак, К. В.; Vorobiev, K V

doi:10.31116/tsitol.2018.04.07

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“…We have also revealed that the pattern of differences in the migratory activity between the cell sublines at the sixth passage at the maximum activity point (4 h after wounding) correlated with the pattern of changes in proliferation activity of the cells at the first passage (proliferation data were obtained previously (Smirnova et al, 2018)). In particular, the enhancement of cell migration and proliferation was observed in the ECV-1ID and ECV-LPS sublines, while enhancement of suppression was observed in the ECV-1000ID subline (Figs.…”

Section: Resultssupporting

confidence: 69%

“…Analysis of cell migration was performed using the previously obtained ECV-304 cell sublines (Smirnova et al, 2018) at the sixth passage after the exposure to influenza virus and/or LPS treatment. Migratory activity was assessed by the change in wound surface area with time (wound sealing).…”

Section: Resultsmentioning

confidence: 99%

“…To detach cells from the surface, a solution containing vercene and chymopsin was used. Both control (intact) and experimental cells were at the sixth passage after the exposure to the agents (Smirnova et al, 2018). Depending on the agent (virus and corresponding ID, LPS, or virus and LPS together), the cell sublines were given the following names: ECV-1ID, ECV-1000ID, ECV-LPS, ECV-1ID + LPS, and ECV-1000ID + LPS.…”

Section: Methodsmentioning

confidence: 99%

“…Normal reproduction of influenza virus is impaired in nonpermissive cells, and the presence of virus is detected only at the level of the viral surface protein mRNA during no more than one or two passages (Danilenko et al, 2016). The change in the physiological state of the cells at the first passage (3-5 days) was the first stage in the changes induced in the cells by influenza virus, with further cell culturing (passages 3 to 8) resulting in the stably increased proliferation at low apoptosis levels (Smirnova et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

“…We have analyzed the changes in the migratory activity of the cells and gene expression at mRNA level, including the genes encoding tumor necrosis factor alpha (TNFα), transforming growth factor beta (TGFβ), and NOX4, as well as the proteins forming part of TJ and controlling the permeability of cell membranes, CLDN5, occludin (OCLD), and ZO-1. Additionally, we assessed the correlation between the functional changes and the proliferation activity and apoptosis in cells of sublines that were analyzed previously (Smirnova et al, 2018), as well as evaluating the ability of rimantadine and alpisarin antiviral preparations to restore the normal functioning of endothelial cells.…”

Section: Introductionmentioning

confidence: 99%

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Long-Term Maintenance of the Functional Changes Induced by Influenza A Virus and/or LPS in Human Endothelial ECV-304 Cell Sublines

et al. 2019

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Influenza A virus and secondary bacterial infection may have remote effects in the form of cardiovascular complications or fibrosis in different organs. However, the mechanisms governing the development of complications remain poorly studied. The present work reports the comparative assessment of the functional changes which take place in human ECV-304 endothelial cell sublines obtained previously by the longterm culturing of cells after exposure to varying infectious doses (IDs) of influenza A virus, and/or bacterial lipopolysaccharide (LPS). It has been demonstrated that, in the course of long-term culturing (six passages) after exposure to pathogenic agents (influenza virus and/or LPS), endothelial cells maintain changes in their migratory activity, permeability, and expression of mRNA for cytokines TNFα and TGFβ (along with the changes in their proliferation activity, which has been demonstrated earlier). The pattern of changes depended on the type of the agent (agents) to which the cells were exposed. The differences in migratory activity (which was at its maximum 4 h after wounding) between the cell sublines at the sixth passage correlated with the differences in their proliferation activity at the first passage (proliferation data were obtained previously). In particular, an increase in migration and proliferation was observed in the sublines exposed to low virus doses (ECV-1ID), as well as exposed to LPS (ECV-LPS), while the suppression of migration and proliferation was observed in the subline exposed to high virus doses (ECV-1000ID). In the ECV-1ID, ECV-LPS, and most notably in ECV-1ID + LPS sublines, we detected an increase in the expression of mRNA for cytokines TNFα and TGFβ, which, however, didn't lead to the induction of apoptosis. We have also demonstrated an increase in cell permeability in the analyzed sublines, which was indicated by a decrease in the expression of the mRNAs for the genes encoding occludin and ZO-1, the tight junctions proteins. This paper also reports an evaluation of the effects of the antiviral preparations rimantadine and alpisarin on the functional state of cell sublines. As a result, it has been demonstrated that these drugs may be able to prevent the development of the pathological changes caused by influenza A virus and/or LPS in endothelial cells. The results obtained in the present work may be of use when studying the mechanisms of development of the influenza A virus and secondary bacterial infection complications.

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Section: Resultssupporting

confidence: 69%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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