Retroviral transduction of a mutant erbB-2 gene into human CD34(+) deriveddendritic cells

Objective To successfully transduce a mutant erbB-2 gene into normal human CD34(+)-derived dendritic cells (DCs) and verify gene expression in these t ransduced dendritic cells. Methods The packaging cell line PA317 was transfected with mutant erbB-2 ge ne DNA and the virus produced was used to infect packaging cell line PG13. The virus produced by PG13 was used to infect CD34+-derived dendritic cells by the spinoculation method using flasks coated with fibronectin material to facilitate retrovirus gene transter efficiency and the mutant erbB-2 gen e expression was assessed by ABC staining and FACScan methods. Results A mutant erbB-2 gene packaging cell line was produced and this muta nt gene was transduced into human CD34(+)-derived DCs. It was verified that the relatively large numbers of the transduced DCs expressed the mutant er bB-2 protein which was eradicated of the ability to transform mouse NIH3T3 fibroblast cells. Conclusions Human DCs can be gene-modified and these gene-modified DCs may be useful in stimulating T lymphocytes for immunotherapy.