Expression of pIX Gene Induced by Transgene Promoter: Possible Cause of Host Immune Response in First-Generation Adenoviral Vectors

Nakai, Michiko; Komiya, Kazuo; Murata, Masashi; Kimura, Tōru; Kanaoka, Masaharu; Kanegae, Yumi; Saito, Izumu

doi:10.1089/hum.2007.085

Cited by 42 publications

(54 citation statements)

References 51 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This virus (virus A, Table 1) induced a significantly greater innate inflammatory response than did the viruses deleted for all of E1 or containing the E1A promoter followed by a poly A site (Fig. 3; P <0.0002 for both pairwise comparisons), in agreement with the results of Nakai et al (2007).…”

Section: Resultssupporting

confidence: 83%

Strong foreign promoters contribute to innate inflammatory responses induced by adenovirus transducing vectors

et al. 2011

View full text Add to dashboard Cite

E1-deleted adenovirus (FG Ad) transducing vectors are limited for use in vivo by their induction of strong innate and adaptive inflammatory responses. We have examined the contribution of the transgene cassette, particularly the foreign promoter driving transgene expression, in the induction of innate inflammation using a mouse ear model in which swelling is measured as a sensitive surrogate marker of the total innate inflammatory response. The commonly used cytomegalovirus major immediate early (CMV) promoter led to high-level swelling that was independent of transgene expression, while the Rous sarcoma virus and human ubiquitin C promoters led to intermediate levels of swelling and the Ad E1A promoter or no promoter led to equally low levels of swelling. Significant swelling was induced by a virus in which the E1A promoter directed pIX expression, supporting the possibility that activation of expression of Ad genes retained in the vector plays an important role in the inflammatory response. Taken together, our findings support the idea that strong foreign promoters likely play the limiting role in the induction of innate and adaptive immune responses that limit the duration of transgene expression after transduction by FG Ad vectors.

show abstract

Section: Resultssupporting

confidence: 83%

Strong foreign promoters contribute to innate inflammatory responses induced by adenovirus transducing vectors

et al. 2011

View full text Add to dashboard Cite

show abstract

“…16 In addition to the forward orientation, the liver-specific expression cassette was inserted in the inverse orientation to eliminate the potential expression of AdV proteins from the LP1 promoter, for instance, due to aberrant splicing. 19 In the Gunn rat, the efficacy of the resulting vectors Ad-LP1-UGT1A1 and Ad-LP1-UGT1A1inv is compared with that of scAAV1-LP1-UGT1A1 and with that of scAAV8-LP1-UGT1A1. Tail vein injection of a dose 3 Â 10 10 ip kg À 1 of Ad-LP1-UGT1A1inv normalizes serum bilirubin levels (Figure 2a).…”

Section: Resultsmentioning

confidence: 99%

“…18,19 Using the weaker but liverspecific LP1 promoter reduces this expression and limits it to hepatocytes. This results in sustained correction.…”

Section: Discussionmentioning

confidence: 99%

“…Prolonged expression with first-generation liver-specific AdV vectors has been reported previously, for instance, in a mouse model for AAT deficiency and of hGH in C57BL/6 mice. 19,26 In the human situation, however, the presence of a memory response against AdV proteins may result in a more rapid and more potent immune response. The higher efficacy of a HD-Ad compared with a liver-specific first-generation AdV vector in an immune competent mouse model suggest that this mechanism also occurs in a naive animal model, albeit to a lesser extent.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

In the rat liver, Adenoviral gene transfer efficiency is comparable to AAV

et al. 2013

View full text Add to dashboard Cite

Adenoviral (AdV) and Adenovirus-associated viral (AAV) vectors both are used for in vivo gene therapy of inherited liver disorders, such as Crigler-Najjar syndrome type 1. In a relevant animal model, the Gunn rat, both vectors efficiently correct the severe hyperbilirubinemia characteristic of this liver disorder. Although the clinical use of AAV is more advanced, as demonstrated by the successful phase 1 trial in hemophilia B patients, because of its large cloning capacity AdV remains an attractive option. A direct comparison of the efficacy of these two vectors in the liver in a relevant disease model has not been reported. Aim of this study was to compare the efficiency of clinically applicable doses of both vectors in the Gunn rat. AdV or scAAV (self-complimentary AAV) ferrying identical liver-specific expression cassettes of the therapeutic gene, UGT1A1, were injected into the tail vein. As the titration methods of these two vectors are very different, a comparison based on vector titers is not valid. Therefore, their efficacy was compared by determining the amount of vector genomes delivered to the liver required for therapeutic correction of serum bilirubin. Like AAV, the liver-specific first-generation AdV also provided sustained correction in this relevant disease model. UGT1A1 mRNA expression provided per genome was comparable for both vectors. Flanking the expression cassette in AdV with AAV-ITRs (inverted terminal repeats), increased UGT1A1 mRNA expression eightfold which resulted in a significant improvement of efficacy. Compared with AAV, less AdV genomes were needed for complete correction of hyperbilirubinemia.

show abstract

“…Juxtaposition of the promoter with the enhancer has been shown to interfere with a controlled transcription of the transgene and is suspected to lead to a low-level expression of the remaining Ad early genes with detrimental effects. 19 Recently, Nakai et al 20 have verified readthrough from the transgene into the adjacent Ad pIX gene even with the production of aberrantly spliced transgene-pIX chimeras, effects that were absent in identical vectors but with leftward orientation of the expression cassette.…”

Section: Co-expression Of P21 In Adenoviral Vector a Schumacher Et Almentioning

confidence: 99%

Co-expression of p21Waf1/Cip1 in adenovirus vectors improves expression of a second transgene

et al. 2009

View full text Add to dashboard Cite

First-generation adenoviral (Ad) vectors are frequently used vectors for experimental and clinical gene transfer. Earlier it has been shown that parallel overexpression of the cell cycle regulator p21 Waf1/Cip1 (p21) or antiapoptotic bcl-2 from a second vector reduces cytotoxicity and improves transgene expression. Here, we investigate whether the coexpression of p21 and a 1 -antitrypsin from a single vector improves vector safety and a 1 -antitrypsin expression. Cell lines (A549 and HeLa) and primary cells (small airway epithelial cells and hepatocytes) were infected with adenovirus vectors transducing a 1 -antitrypsin with (AdCMV.p21-RSV.hAAT) or without (AdRSV.hAAT) p21. a 1 -Antitrypsin expression and cytotoxicity were analyzed using western blot/ELISA and LDH/ALT/AST assays, respectively. Cell cycle profiles were determined by flow cytometry. Coexpression of p21 strongly increased the a 1 -antitrypsin expression in all cell types and at all doses tested. No changes in ALT/AST from hepatocytes and only minor increases in the LDH release in A549 and HeLa were observed with either vector. Cell cycle profiles were also not affected adversely. Incorporation of p21 in Ad vectors together with a gene of interest improves the vector performance; such vectors will allow the application of lower doses and thereby reduce immunological side effects.

show abstract

Expression of pIX Gene Induced by Transgene Promoter: Possible Cause of Host Immune Response in First-Generation Adenoviral Vectors

Cited by 42 publications

References 51 publications

Strong foreign promoters contribute to innate inflammatory responses induced by adenovirus transducing vectors

Strong foreign promoters contribute to innate inflammatory responses induced by adenovirus transducing vectors

In the rat liver, Adenoviral gene transfer efficiency is comparable to AAV

Co-expression of p21Waf1/Cip1 in adenovirus vectors improves expression of a second transgene

Contact Info

Product

Resources

About