Objective To successfully transduce a mutant erbB 2 gene into normal human CD34 + derived dendritic cells (DCs) and verify gene expression in these transduced dendritic cells Methods The packaging cell line PA31...Objective To successfully transduce a mutant erbB 2 gene into normal human CD34 + derived dendritic cells (DCs) and verify gene expression in these transduced dendritic cells Methods The packaging cell line PA317 was transfected with mutant erbB 2 gene 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 gene expression was assessed by ABC staining and FACScan methods Results A mutant erbB 2 gene packaging cell line was produced and this mutant gene was transduced into human CD34 + derived DCs It was verified that the relatively large numbers of the transduced DCs expressed the mutant erbB 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展开更多
文摘Objective To successfully transduce a mutant erbB 2 gene into normal human CD34 + derived dendritic cells (DCs) and verify gene expression in these transduced dendritic cells Methods The packaging cell line PA317 was transfected with mutant erbB 2 gene 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 gene expression was assessed by ABC staining and FACScan methods Results A mutant erbB 2 gene packaging cell line was produced and this mutant gene was transduced into human CD34 + derived DCs It was verified that the relatively large numbers of the transduced DCs expressed the mutant erbB 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
