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Pneumococcus Infection of Primary Human Endothelial Cells in Constant Flow
Published on: October 31, 2019
Long-Term Maintenance of the Functional Changes Induced by Influenza A Virus and/or LPS in Human Endothelial ECV-304
S S Smirnova1, M M Pisareva2, T D Smirnova2
11Peter the Great St. Petersburg Polytechnic University, 195251 St. Petersburg, Russia.
Abstract:
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 long-term 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.
Insights
Influenza A virus and LPS exposure alter endothelial cell function, impacting migration and permeability. Antiviral drugs like rimantadine and alpisarin show potential in preventing these pathological changes, offering insights into infection complications.
Area of Science:
- Endothelial cell biology
- Infectious disease mechanisms
- Cardiovascular complications
Background:
- Influenza A virus and bacterial infections can lead to remote cardiovascular and organ fibrosis complications.
- The underlying mechanisms of these post-infection complications are not well understood.
- Endothelial cells play a crucial role in vascular integrity and response to pathogens.
Purpose of the Study:
- To comparatively assess functional changes in human endothelial cells (ECV-304) after exposure to influenza A virus and/or bacterial lipopolysaccharide (LPS).
- To investigate the long-term effects of varying infectious doses (IDs) of influenza A virus and LPS on endothelial cell behavior.
- To evaluate the potential of antiviral drugs (rimantadine, alpisarin) in mitigating infection-induced endothelial cellpathology.
Main Methods:
- Long-term culturing of human ECV-304 endothelial cell sublines.
- Exposure to different infectious doses of influenza A virus and/or bacterial LPS.
- Assessment of cell proliferation, migration, permeability, and mRNA expression of TNFα, TGFβ, occludin, and ZO-1.
- Evaluation of antiviral drug effects on treated cell sublines.
Main Results:
- Endothelial cells maintained altered migratory activity, permeability, and cytokine mRNA expression after six passages post-exposure.
- Low influenza A virus ID (ECV-1ID) and LPS (ECV-LPS) increased migration and proliferation; high virus ID (ECV-1000ID) suppressed them.
- Increased TNFα and TGFβ mRNA expression observed in ECV-1ID, ECV-LPS, and ECV-1ID + LPS sublines, without inducing apoptosis.
- Increased cell permeability correlated with decreased expression of tight junction proteins (occludin, ZO-1).
- Rimantadine and alpisarin demonstrated potential in preventing pathological changes in endothelial cells.
Conclusions:
- Long-term exposure to influenza A virus and/or LPS induces persistent functional changes in endothelial cells, affecting migration, proliferation, and permeability.
- The observed changes in endothelial cells may contribute to the development of post-infectious complications.
- Antiviral agents rimantadine and alpisarin may serve as protective agents against influenza A virus and LPS-induced endothelial dysfunction.
