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Development of a novel adenovirus–alphavirus hybrid vector with RNA replicon features for malignant hematopoietic cell transduction

Cancer Gene Therapy volume 20, pages 429–436 (2013)Cite this article

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Abstract

To improve the expression levels of transgenes in malignant hematopoietic cells, we developed a novel adenoviral-alphavirus hybrid vector Ad5/F11p-SFV-GFP that contains a Semliki Forest Virus (SFV) replicon and chimeric fibers of Ad5 and Ad11p. Ad5/F11p-SFV-GFP infected >95% of K562, U937 or Jurkat cells and 23.65% of HL-60 cells, and led to moderate Enhanced Green Fluorescent Protein (EGFP) transgene expression intensity. The infection efficiency of Ad5/F11p-SFV-GFP in primary human leukemia cells ranged from 9.34–89.63% (median, 28.58%) at a multiplicity of infection (MOI) of 100, compared with only 3.37–44.54% (median, 10.42%) in cells infected by Ad5/F11p-GFP. Importantly, Ad5/F11p-SFV-GFP led to a significantly higher transgene expression level in primary leukemia cells, as indicated by the relative fluorescence intensity, compared to cells infected with Ad5/F11p-GFP. The increased expression of EGFP in Ad5/F11p-SFV-GFP-infected cells was associated with the accumulation of abundant subgenomic mRNA. Additionally, infection of K562, U937 or Jurkat cells by Ad5/F11p-SFV-GFP was significantly inhibited by blocking CD46 receptor; however, other factors may affect the gene-transfer efficiency of Ad5/F11p-SFV-GFP in primary leukemia cells. In conclusion, we successfully developed a novel adenoviral-alphavirus hybrid vector with RNA replicon features, which represents a promising vector for gene modifications during the production of cell-based vaccines for leukemia patients.

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Acknowledgements

This work was supported by grants from the Chinese National Basic Research and Development (973 Program) (no. 2012CB518205), the National Natural Science Foundation of China (nos. 30901379 and 81170460), and the National High Technology Research and Development Program of China (863 Program) (no. 2012AA02A211).

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  1. Y Yang and F Xiao: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing, People’s Republic of China

    Y Yang, F Xiao, Z Li, Q Zhang, Q Li, H Wang & L-S Wang

  2. Chinese Center for Disease Control and Prevention, Institute for Viral Disease Control and Prevention, Beijing, People’s Republic of China

    Z Lu

  3. Beijing 307 Hospital, The Academy of Military Medical Sciences, Beijing, People’s Republic of China

    H Zuo

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  1. Y Yang
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Correspondence to H Wang or L-S Wang.

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Yang, Y., Xiao, F., Lu, Z. et al. Development of a novel adenovirus–alphavirus hybrid vector with RNA replicon features for malignant hematopoietic cell transduction. Cancer Gene Ther 20, 429–436 (2013). https://doi.org/10.1038/cgt.2013.37

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  • DOI: https://doi.org/10.1038/cgt.2013.37

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