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Isolation of Viral Replication Compartment-enriched Sub-nuclear Fractions from Adenovirus-infected Normal Human Cells
Published on: November 12, 2015
Adeno-associated virus type 5 exploits two different entry pathways in human embryo fibroblasts
Ursula Bantel-Schaal1, Ilona Braspenning-Wesch1, Juergen Kartenbeck2
1German Cancer Research Center, Infection and Cancer Research Program, Im Neuenheimer Feld 242, 69120 Heidelberg, Germany.
The Journal of General Virology
|January 15, 2009
Summary
Adeno-associated virus type 5 (AAV5) uses two distinct pathways to enter human cells: the known clathrin-dependent route and a novel caveolae-mediated pathway. This finding is crucial for understanding gene therapy vector delivery.
Area of Science:
- Virology
- Cell Biology
- Gene Therapy
Background:
- Adeno-associated viruses (AAVs) are promising vectors for gene therapy.
- Understanding AAV uptake and intracellular trafficking is critical for efficient gene delivery.
Purpose of the Study:
- To investigate the entry pathways of adeno-associated virus type 5 (AAV5) in primary human embryo fibroblasts.
- To elucidate the mechanisms governing AAV5 cellular internalization.
Main Methods:
- Microscopy to observe virion localization at low temperatures.
- Analysis of AAV5 uptake at physiological temperatures (37°C).
- Investigation of cellular entry pathways involved in AAV5 internalization.
Main Results:
- AAV5 virions accumulate at cell-cell contact points and filopodia at low temperatures.
- Significant dissociation (up to 80%) of bound AAV5 occurs upon warming to 37°C.
- AAV5 utilizes both clathrin-dependent endocytosis and a parallel pathway involving caveolae and caveosome-like organelles for entry.
Conclusions:
- AAV5 employs a dual-entry mechanism involving both established and novel pathways.
- Caveolar endocytosis represents a previously undescribed route for parvovirus entry.
- These findings have implications for optimizing AAV-based gene therapy strategies.
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