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Pastore et al. [124] injected C3/HeJ mice with first-generation E1-deleted Ad vectors expressing human α1-antitrypsin from either the nonspecific PGK promoter or the liver-specific albumin promoter. In mice that received the phosphoglycerate kinase (PGK) promoter vector, AAT expression declined precipitously within the first 3 weeks, and the mice developed antibodies to AAT. However, in mice that received the albumin promoter vector, AAT expression persisted for greater than 40 weeks, and the mice did not develop antiAAT antibodies [124].

Using this system, Yant et al. [19] con- 20 Yew and Cheng Figure 1 Strategies to increase the stability of transduced vector genomes. Boxes indicate different sequence elements, lines indicate vector backbone sequence, which could be either plasmid or virus. EBV, Epstein–Barr virus; Pr, promoter; pA, polyadenylation signal; FR, EBV family of repeats; SB, Sleeping Beauty; IR/DR, inverted repeats/direct repeats; ITR, inverted terminal repeats. structed a plasmid containing an α-1-antitrypsin (AAT) expression cassette flanked by two ITRs and a second plasmid that expressed the Sleeping Beauty transposase from the cytomegalovirus (CMV) promoter (Fig.

IFN-γ and TNF-α were also shown to act synergistically, further depressing activity [59]. Administration of either adenoviral vectors or cationic lipid–pDNA complexes induces a transient inflammatory response that includes the induction of IFN-γ and TNF-α [60]. These elevated cytokine levels precede the loss of transgene expression. However, IFN-γ and TNF-α levels fall to basal levels within a few days, whereas expression from the CMV promoter remains depressed. Thus, if cytokines were principally responsible for the inactivation of the CMV promoter, their effects would have to be irreversible.

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