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Conditional cell suicide using dox‐dependent caspase‐2 expression
Author(s) -
Knott Anja,
Müller Yvonne,
Zunino Susan J.,
Berens Christian,
Hillen Wolfgang
Publication year - 2003
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.342
Subject(s) - programmed cell death , suicide gene , apoptosis , biology , thymidine kinase , transfection , cell , hela , cell culture , microbiology and biotechnology , cancer research , immunology , genetic enhancement , gene , genetics , herpes simplex virus , virus
Background Adoptive immune transfer is used as an efficient treatment modality to achieve a graft‐versus‐leukemia effect in persisting or relapsing residual leukemic disease. Safety considerations dictate the need for equipping the transferred cells with a conditional suicide mechanism to eliminate donor T cells when graft‐versus‐host disease occurs. We have examined in a model system using HeLa cells whether doxycycline (dox)‐dependent expression of pro‐apoptotic proteins could be used as a potential new strategy for conditional cell elimination. Methods Four constructs encoding pro‐apoptotic proteins were tested in transient transfections to identify suitable cell death inducers. Murine caspase‐2 placed under Tet‐control was chosen for stable transfection into cell lines carrying different dox‐dependent transregulators. The efficiency of cell death induction and the expression patterns of caspase‐2 were analyzed in the respective clones. Results Different levels of induced cell death were obtained depending on the properties of the transregulators used to control target gene expression. High expression levels of caspase‐2 in the presence of dox were required to achieve efficient induction of cell death, while tight repression in the absence of inducer was not necessary for cell survival. Dox treatment for 48 h resulted in 94% cell death indicating a very efficient conditional suicide mechanism. Conclusions We propose that the principle of using pro‐apoptotic cellular proteins placed under appropriate dox‐dependent regulation may represent an alternative conditional suicide mechanism to the frequently used herpes simplex virus thymidine kinase (HSV‐TK)/ganciclovir‐system, which harbors immunological and toxicological risks. Copyright © 2002 John Wiley & Sons, Ltd.