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Transduction of CD34+ cells by a vesicular stomach virus protein G (VSV-G) pseudotyped HIV-1 vector. Stable gene
1Department of Medicine, University of California, San Diego, La Jolla, USA. fwongstaal@ucsd.edu
Objective:
To use HIV-1 vectors to mediate stable gene transfer into hematopoietic stem/progenitor cells.
Study Design/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.