Efficient gene transfer into hematopoietic cells by a retargeting adenoviral vector system with a chimeric fiber of adenovirus serotype 5 and 11p.
OBJECTIVE
Adenoviral vectors (Ad) were widely used in gene therapy and study of gene function, but the commonly used serotype 5 adenovirus-based vectors (Ad5) could poorly transduce hematopoietic cells because of low expression of viral receptors on these cells. To overcome this limitation, we developed a retargeting adenovector with a chimeric fiber of Ad5 and Ad11p (Ad5F11p) and evaluated its gene transfer ability in hematopoietic cells.


METHODS
An Ad11p fiber pseudotyped Ad5 vector was generated by modifying the fiber gene of pAdEasy-1 backbone plasmid. Ad5F11p-GFP encoding enhanced green fluorescence protein (GFP) gene was transferred into human leukemic cell lines, primary leukemic cells, and CD34(+) hematopoietic cells. The gene transduction efficiency was determined by fluorescence-activated cell sorting assay.


RESULTS
More than 90% of U937 or K562 cells could be infected by Ad5F11p-GFP at a moderate multiplicity of infection (MOI). Ad5F11p-GFP is also significantly more effective than control Ad5-GFP in infection of primary myeloid leukemic cells. At 200 MOI, GFP-positive percentages of Ad5F11p-GFP transduced myeloid leukemic cells range from 10.58% to 92.63% with a median of 28.65%. Ad5F11p-GFP could transduce about 50% human hematopoietic stem/progenitor (CD34(+)) cells, while Ad5-GFP could transduce <15% at 200 MOI. CD46 was reported to be the receptor of Ad11p. Our data suggest that CD46 participates in the process of Ad5F11p-GFP infection but is not the unique molecule determining its gene transfer efficiency of host cells.


CONCLUSIONS
We established a retargeting adenovector system, which could infect hematopoietic cells effectively and would benefit research work on Ad tropism.
Lu
Zhuo-Zhuang
ZZ
Department of Experimental Hematology, Beijing Institute of Radiation Medicine, Beijing, P.R. China.
Ni
Fang
F
Hu
Ze-Bin
ZB
Wang
Lan
L
Wang
Hua
H
Zhang
Qun-Wei
QW
Huang
Wen-Rong
WR
Wu
Chu-Tse
CT
Wang
Li-Sheng
LS
D000256:Adenoviridae; D018952:Antigens, CD34; D045744:Cell Line, Tumor; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006412:Hematopoietic Stem Cells; D006801:Humans; D013045:Species Specificity; D014161:Transduction, Genetic / Q000379:methods; D019076:Transgenes
D018952:Antigens, CD34; D049452:Green Fluorescent Proteins