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Dissection of Adult Mouse Utricle and Adenovirus-mediated Supporting-cell Infection
Published on: March 28, 2012
Ultrastructural and immunofluorescence studies of early events in adenovirus-HeLa cell interactions
Abstract:
Scanning electron microscopy and immunofluorescence techniques show that the attachment of adenovirus type 5 to HeLa cells is followed by a temperature-dependent redistribution of virus particles on the cell surface. Metabolic inhibitors and cytochalasin B, a drug that impairs microfilament function, blocked this redistribution. Transmission electron microscopy studies demonstrated that inhibition of redistribution was paralleled by an inhibition of virus internalization. In further experiments virus and cells were incubated at 4 degrees C and then treated with adenovirus 'soluble' antigens to block unoccupied virus receptors. On warming these preparations to 37 degrees C, the internalization of attached virions was found to be impaired. It is proposed that energy-dependent redistribution of attached adenovirus particles on the cell surface facilitates entry by bringing virions into contact with sufficient receptors to allow internalization.
Insights
Adenovirus type 5 entry into HeLa cells requires virus particle redistribution on the cell surface. This energy-dependent process, crucial for viral internalization, can be blocked by metabolic inhibitors and cytochalasin B.
Area of Science:
- Cell Biology
- Virology
- Microscopy Techniques
Background:
- Adenovirus type 5 (Ad5) is a significant human pathogen.
- Understanding the initial steps of viral entry is crucial for developing antiviral strategies.
Purpose of the Study:
- To investigate the role of cell surface dynamics in adenovirus type 5 internalization into HeLa cells.
- To elucidate the mechanisms governing virus-cell interactions during entry.
Main Methods:
- Scanning electron microscopy (SEM) and immunofluorescence for visualizing virus distribution.
- Transmission electron microscopy (TEM) for ultrastructural analysis of internalization.
- Use of metabolic inhibitors and cytochalasin B to probe cellular processes.
- Blocking experiments with soluble adenovirus antigens.
Main Results:
- Ad5 attachment to HeLa cells induced temperature-dependent redistribution of virus particles.
- Metabolic inhibitors and cytochalasin B blocked this redistribution and subsequent virus internalization.
- Impaired redistribution at low temperatures or with blocked receptors reduced viral entry.
Conclusions:
- Energy-dependent redistribution of Ad5 on the cell surface is essential for efficient viral internalization.
- This redistribution likely facilitates entry by promoting contact with sufficient cellular receptors.
- Microfilament function plays a role in the Ad5 entry process.
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