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[Retroviral transduction of a mutant erbB-2 gene into human CD34+ derived dendritic cells]
1Research Laboratory of Blood Physiology, Hunan Medical University, Changsha 410078, China.
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.