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The relationship between vector dose and AADC enzymatic activity in tissue extracts was linear.
We show that IgG expression level can be maximized as a function of vector dose and transgene ratio.
Hepatic insulin production resulted in the reduction of blood glucose in treated diabetic rats, the degree of blood glucose reduction correlated with both the vector dose and the level of hepatic insulin expression.
We report here the results of an investigation of the relationship between vector dose and a series of efficacy markers, such as PET, l-Dopa response, and AADC enzymatic activity.
The benchmarking highlights an agglomeration of the free PEI molecules into micron sized clusters and identifies the metric determining cell death as the total number of PEI molecules presented to cells, determined by the delivery vector dose and the surface density of the cargo.
Although our data show a mild activation of unfolded protein response markers following F8 gene delivery at a certain vector dose in C57BL/6 mice, it was not augmented upon further elevated dosing, did not induce liver pathology or apoptosis, and did not impact FVIII immunogenicity.
A reduction in vector dose resulted in reduced levels of transcripts (data not shown).
This strategy potentially allows a reduction of the injected vector dose as compared to an injection of AAV2 alone.
Increasing the vector dose or adopting a prime-boost regimen in order to overcome pre-existing immunity to the virus is a common approach [2].
In any of these approaches, predicting the vector dose that optimizes safety, particularly in vector-naïve individuals, yet provides optimal immunogenicity will facilitate the design of clinical trials.
We observed a direct correlation between vector dose and cytokine expression indicating that lentiviral based cytokine delivery can be easily adjusted to deliver a desired dose of cytokine.
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