Retroviral transduction of a mutant erbB-2 gene into human CD34~+ derived dendritic cells

来源 :Chinese Medical Journal | 被引量 : 0次 | 上传用户:homking14
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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 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 transmitted 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 th ese gene modified DCs may be useful in stimulating T lymphocytes for immunotherapy
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