title: Development of a novel adenovirus-alphavirus hybrid vector with RNA replicon features for malignant hematopoietic cell transduction.
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.
authors.affiliation: Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing, People's Republic of China.
authors.forename: Y
authors.initials: Y
authors.lastname: Yang
authors.forename: F
authors.initials: F
authors.lastname: Xiao
authors.forename: Z
authors.initials: Z
authors.lastname: Lu
authors.forename: Z
authors.initials: Z
authors.lastname: Li
authors.forename: H
authors.initials: H
authors.lastname: Zuo
authors.forename: Q
authors.initials: Q
authors.lastname: Zhang
authors.forename: Q
authors.initials: Q
authors.lastname: Li
authors.forename: H
authors.initials: H
authors.lastname: Wang
authors.forename: L-S
authors.initials: LS
authors.lastname: Wang
mesh_terms: D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D000524:Alphavirus / Q000235:genetics* / Q000378:metabolism; D045744:Cell Line, Tumor; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D018922:HL-60 Cells; D006412:Hematopoietic Stem Cells / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D019169:Jurkat Cells; D020014:K562 Cells; D007938:Leukemia / Q000821:virology; D010957:Plasmids / Q000235:genetics; D012367:RNA, Viral / Q000235:genetics*; D012093:Replicon / Q000235:genetics*; D012672:Semliki forest virus / Q000235:genetics; D014161:Transduction, Genetic; D019076:Transgenes
chemical_list: D012367:RNA, Viral; D049452:Green Fluorescent Proteins