Development of an adenoviral vector system with adenovirus serotype 35 tropism; efficient transient gene transfer into primary malignant hematopoietic cells.
BACKGROUND
A paucity of coxsackie adenovirus receptor (CAR) hampers the adenovirus serotype 5 (Ad5)-based vector-mediated gene transfer into malignant hematopoietic cells. Fiber-retargeted adenoviral vectors with species B tropism can potentially bypass the CAR requirement and facilitate efficient gene transfer into malignant hematopoietic cells.


METHODS
For feasible generation of fiber-retargeted adenoviral vectors, we have modified the versatile AdEasy system with a chimeric fiber gene encoding the Ad5 fiber tail domain and Ad35 fiber shaft and knob domains. An Ad5-based vector encoding the green fluorescent protein (GFP) gene under the control of the PGK promoter with Ad35 fiber receptor specificity was generated (Ad5F35-GFP). The Ad5F35-GFP vector-mediated gene transfer efficiency was compared with a fiber non-modified Ad5-GFP vector, which also encodes the GFP gene under the control of the PGK promoter.


RESULTS
We demonstrated that a variety of Ad5-refractory malignant myeloid and B lymphoid cell lines were highly permissive to the Ad5F35-GFP vector infection. Importantly, primary chronic myeloid leukemic (CML) cells and chronic lymphocytic leukemia (CLL) B cells were superiorly transduced by the Ad5F35-GFP vector at a multiplicity of infection (MOI) of 100 compared with the Ad5-GFP vector.


CONCLUSIONS
Our study will facilitate the generation of fiber-retargeted adenoviral vectors and enable transient genetic manipulation of primary malignant hematopoietic cells.
Nilsson
Marcus
M
Department of Molecular Medicine and Gene Therapy, Lund University, SE-22184 Lund, Sweden.
Ljungberg
Johan
J
Richter
Johan
J
Kiefer
Thomas
T
Magnusson
Mattias
M
Lieber
André
A
Widegren
Bengt
B
Karlsson
Stefan
S
Fan
Xiaolong
X
D000256:Adenoviridae / Q000235:genetics*; D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006413:Hematopoietic System / Q000473:pathology*; D039564:Integrin alphaV / Q000378:metabolism; D015448:Leukemia, B-Cell / Q000235:genetics / Q000473:pathology / Q000628:therapy; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000235:genetics / Q000473:pathology / Q000628:therapy; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics / Q000473:pathology / Q000628:therapy; D010735:Phosphoglycerate Kinase / Q000235:genetics; D011991:Receptors, Virus / Q000235:genetics / Q000378:metabolism; D012703:Serotyping; D014407:Tumor Cells, Cultured; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
D062727:Coxsackie and Adenovirus Receptor-Like Membrane Protein; D039564:Integrin alphaV; D011991:Receptors, Virus; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; D010735:Phosphoglycerate Kinase