Transduction of CD34+ cells by a vesicular stomach virus protein G (VSV-G) pseudotyped HIV-1 vector. Stable gene expression in progeny cells, including dendritic cells.
OBJECTIVE
To use HIV-1 vectors to mediate stable gene transfer into hematopoietic stem/progenitor cells.


METHODS
Purified human CD34+ cells were transduced with HIV-1 vectors pseudotyped with VSV-G and subjected to colony-forming assays and differentiation in liquid culture. Transduction was determined by DNA-polymerase chain reaction (PCR) for the transgene. GFP reporter gene expression and phenotypes of progeny cells were assessed by microscopy and flow cytometry.


RESULTS
The HIV-1 vector transduced CD34+ cells with high efficiency. Transduction did not interfere with CD34+ cells differentiation in vitro. Transduced genes are expressed in different subsets of progeny cells, including those with normal dendritic cells (DC) morphology and phenotypes (HLADR+/CD1a+/CD86+/CD14-).


CONCLUSIONS
We have demonstrated efficient transduction of hematopoietic progenitor cells by HIV-1 vectors. The transgenes are expressed in different subsets of progeny cells, which suggests stable integration. The generation of DCs stably expressing HIV antigens provides a new approach for vaccine development.
Li
X
X
Department of Medicine, University of California, San Diego, La Jolla, USA. fwongstaal@ucsd.edu
Mukai
T
T
Young
D
D
Frankel
S
S
Law
P
P
Wong-Staal
F
F
D000818:Animals; D018952:Antigens, CD34 / Q000235:genetics*; D019556:COS Cells; D002454:Cell Differentiation; D003713:Dendritic Cells / Q000166:cytology / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000235:genetics*; D015870:Gene Expression / Q000235:genetics; D017930:Genes, Reporter / Q000235:genetics; D005814:Genes, Viral / Q000235:genetics; D005822:Genetic Vectors / Q000008:administration & dosage; D015497:HIV-1 / Q000235:genetics* / Q000276:immunology; D006412:Hematopoietic Stem Cells / Q000821:virology*; D006801:Humans; D007963:Leukocytes, Mononuclear; D014162:Transfection / Q000379:methods*; D014721:Vesicular stomatitis Indiana virus / Q000235:genetics*
D018952:Antigens, CD34; D019204:GTP-Binding Proteins