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Exposure to exogenous DNA can modify the sensitivity of the Fas apoptotic pathway
Author(s) -
de Carvalho Bittencourt Marcelo,
Saas Philippe,
Fresnay Stéphanie,
YerlyMotta Véronique,
Ferrand Christophe,
Perruche Sylvain,
Duperrier Anne,
Hervé Patrick,
Tiberghien Pierre,
Chalmers David E.
Publication year - 2001
Publication title -
the journal of gene medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.689
H-Index - 91
eISSN - 1521-2254
pISSN - 1099-498X
DOI - 10.1002/jgm.226
Subject(s) - fas ligand , apoptosis , transfection , biology , programmed cell death , cationic liposome , microbiology and biotechnology , cytotoxic t cell , dna fragmentation , fas receptor , naked dna , cell culture , chemistry , in vitro , biochemistry , genetics
Abstract Background Gene‐transfer techniques are commonly employed for both in vitro and in vivo studies. However, modifications of the target cell following the introduction of the gene of interest are not often examined. These modifications can alter the immunogenicity and/or the susceptibility of the target cell to apoptosis and may produce unwanted consequences in vivo . Methods Gene transfer into the murine fibroblastic Ψ‐CRIP packaging cellline was performed using calcium phosphate precipitation, cationic liposome–DNA complexes or a retroviral RNA‐mediated method. After gene transfer, Fas expression, cytokine production, and sensitivity to Fas ligand (FasL)‐mediated death were assessed. Results Following transfection of a FasL expression vector by calcium phosphate precipitation, an unexpected increase was observed in apoptotic cell death in previously Fas‐resistant Ψ‐CRIP cells. This apoptosis was due to Fas upregulation and an increase of sensitivity to FasL‐mediated death. Other plasmids coding non‐cytotoxic factors also modulated this apoptotic pathway. Theco‐stimulatory molecule CD80 was also upregulated. Exposure to naked DNA alone elicited the same response. The effect was not dependent on the methylation status of exogenous DNA, but was found to be dependent on the target cell type and might be avoided by the use of an RNA‐mediated retroviral system. Conclusions Plasmid transfection or simple exposure to naked DNA can increase sensitivity to apoptosis. The generation of FasL packaging cell lines is therefore limited by an increase in FasL/Fas‐mediated apoptosis. These findings should be considered when using genetically modified transplantable cells in order to prevent elimination by host cytotoxic cells and in particular when cells are engineered using FasL. Copyright © 2001 John Wiley & Sons, Ltd.