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Published on: August 20, 2015
Liposome-mediated transient transfection reduces cholesterol-dependent coxsackievirus infectivity
Jerry Wong1, Jingchun Zhang, Guang Gao
1Department of Pathology and Laboratory Medicine, The James Hogg iCAPTURE Centre for Cardiovascular and Pulmonary Research, University of British Columbia-St. Paul's Hospital, Vancouver, Canada.
Abstract:
Liposome-mediated gene delivery provides a powerful strategy for the study of gene function and for gene therapy. Coxsackievirus B3 is an important human pathogen associated with various diseases. Here we reported that liposome-mediated transient transfection of plasmid cDNA inhibited coxsackieviral replication at the levels of RNA, protein and viral progeny release. These inhibitory effects were observed in various cell types and by using different liposome reagents. We further showed that the inhibition was likely due to the lack of virus attachment. Moreover, we showed that addition of cholesterol restored, at least in part, the viral infectivity. Interestingly, we found that membrane cholesterol levels were unchanged during transfection, indicating that disruption rather than depletion of membrane cholesterol contributes to the inhibitory effects of transfection. Our data suggest that liposome-mediated cDNA transient transfection inhibits coxsackievirus infectivity via inhibition of viral attachment, which is likely occurring through the changes of membrane cholesterol integrity.
Insights
Liposome-mediated gene delivery inhibits coxsackievirus B3 replication by preventing viral attachment. This effect, observed across cell types, is linked to membrane cholesterol integrity changes, not depletion.
Area of Science:
- Molecular Biology
- Virology
- Biotechnology
Background:
- Liposome-mediated gene delivery is crucial for gene function studies and gene therapy.
- Coxsackievirus B3 is a significant human pathogen causing diverse diseases.
Purpose of the Study:
- To investigate the impact of liposome-mediated transient transfection on Coxsackievirus B3 replication.
- To elucidate the mechanism underlying the observed inhibition of viral replication.
Main Methods:
- Liposome-mediated transient transfection of plasmid cDNA in various cell types.
- Assessment of viral RNA, protein, and progeny release.
- Analysis of viral attachment and membrane cholesterol levels.
Main Results:
- Transient transfection significantly inhibited Coxsackievirus B3 replication at multiple levels.
- Inhibition was associated with impaired viral attachment.
- Cholesterol addition partially restored infectivity, suggesting membrane integrity is key.
- Transfection did not alter overall membrane cholesterol levels, but disrupted its integrity.
Conclusions:
- Liposome-mediated transfection inhibits Coxsackievirus B3 infectivity by disrupting membrane cholesterol integrity, thereby preventing viral attachment.
- This finding offers insights into viral entry mechanisms and potential therapeutic strategies.
