The DOI matches its Crossref record; 56 of the 56 references it cites were found in a registry or an index.
Efficient Gene Transfer into Human CD34 + Cells by a Retargeted Adenovirus Vector
Open accessAbstract
Efficient infection with adenovirus (Ad) vectors based on serotype 5 (Ad5) requires the presence of coxsackievirus-adenovirus receptors (CAR) and α v integrins on cells. The paucity of these cellular receptors is thought to be a limiting factor for Ad gene transfer into hematopoietic stem cells. In a systematic approach, we screened different Ad serotypes for interaction with noncycling human CD34 + cells and K562 cells on the level of virus attachment, internalization, and replication. From these studies, serotype 35 emerged as the variant with the highest tropism for CD34 + cells. A chimeric vector (Ad5GFP/F35) was generated which contained the short-shafted Ad35 fiber incorporated into an Ad5 capsid. This substitution was sufficient to transplant all infection properties from Ad35 to the chimeric vector. The retargeted, chimeric vector attached to a receptor different from CAR and entered cells by an α v integrin-independent pathway. In transduction studies, Ad5GFP/F35 expressed green fluorescent protein (GFP) in 54% of CD34 + cells. In comparison, the standard Ad5GFP vector conferred GFP expression to only 25% of CD34 + cells. Importantly, Ad5GFP transduction, but not Ad5GFP/F35, was restricted to a specific subset of CD34 + cells expressing α v integrins. The actual transduction efficiency was even higher than 50% because Ad5GFP/F35 viral genomes were found in GFP-negative CD34 + cell fractions, indicating that the cytomegalovirus promoter used for transgene expression was not active in all transduced cells. The chimeric vector allowed for gene transfer into a broader spectrum of CD34 + cells, including subsets with potential stem cell capacity. Fifty-five percent of CD34 + c-Kit + cells expressed GFP after infection with Ad5GFP/F35, whereas only 13% of CD34 + c-Kit + cells were GFP positive after infection with Ad5GFP. These findings represent the basis for studies aimed toward stable gene transfer into hematopoietic stem cells.
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Cited by
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Metadata
- Authors
- Shayakhmetov, D. M., Papayannopoulou, T., Stamatoyannopoulos, G., & Lieber, A.
- Venue
- Journal of Virology
- Publisher
- American Society for Microbiology
- Published
- 2000-03-15
- Volume
- 74
- Issue
- 6
- Pages
- 2567-2583
- ISSN
- 0022-538X (ISSN-L)
- 1098-5514
- OpenAlex
- W2162380590
- Cited by
- 367
- References
- 56
- FWCI
- 15.03
- Type
- journal article
- Language
- en
- Open access
- green
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- Shayakhmetov, D. M., Papayannopoulou, T., Stamatoyannopoulos, G., & Lieber, A. (2000). Efficient Gene Transfer into Human CD34 + Cells by a Retargeted Adenovirus Vector. Journal of Virology. https://doi.org/10.1128/jvi.74.6.2567-2583.2000
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