IL12-Mediated Liver Inflammation Reduces the Formation of AAV Transcriptionally Active Forms but Has No Effect over Preexisting AAV Transgene Expression
Figure 2
IL-12 down-regulates AAV mediated gene expression without reducing the number of viral genomes.
C57BL/6 mice (n = 5) received 2.5×1011 vg/kg of AAV8-luc vector alone or in combination with 1.5×109 vg/kg of AAV8-IL12 vector. A) Luciferase expression levels were analyzed in both groups of animals at days 1, 4, 7, and 14 using a Xenogen in vivo luminometer. Luciferase expression was quantified and represented as photons/second. The data are shown as mean ± standard deviation (SD). The differences in luciferase expression were statistically evaluated by Student t test (**p<0.01). B) Animals were sacrificed at day 14. Total DNA and RNA were isolated from liver and vector genome copies and luciferase messenger RNA (mRNA) copies were determined using a q-PCR specific for luciferase, normalized to GADPH copy numbers. Values correspond to the mean copy number from five animals. C) The levels of luciferase activity (relative light units, RLUs) were determined in liver homogenates and were normalized to protein content (RLU/mg). The data are shown as mean ± SEM. D) Serum transaminases were analyzed 14 days after vector injection. The data are shown as mean ± standard deviation (SD). Each experiment was repeated three times.