Figure 9.

Liver untargeting of Ad infection using a bispecific adapter, sCAR-CXCL12, in SCID-bg mice in vivo.

Notes: Ad5-CMV-GFP-luc (1 × 10⁸ ifu/mouse) pre-incubated in the absence or presence of sCAR-CXCL12 (5 μg) was injected intravenously into SCID-bg mice. Ad-directed luc expression was monitored in living animals by bioluminescence imaging at 3 days after injection. (A) Examples of an untreated mouse, a mouse injected with Ad5-CMV-GFP-luc, and a mouse injected with Ad5-CMV-GFP-luc + sCAR-CXCL12. Luc expression was quantified from ROI of the liver regions. (B) Mean count ± SEM from ten animals. After bioluminescent imaging, the mice were sacrificed, liver extracts were prepared, and Ad was quantified by real-time PCR. (C) Mean E4 copy number ± SEM in livers from ten animals. The treatment results were compared using a Student’s t-test; the differences were considered statistically significant (*) if p<0.05. Abbreviations: Ad, adenovirus; Ad5, Ad serotype 5; GFP, green fluorescent protein; ifu, infectious unit; luc, luciferase; PCR, polymerase chain reaction; ROI, regions of interest; sCAR, soluble ectodomain form of the native Ad5 receptor; SCID-bg, severe combined immunodeficiency-beige; SEM, standard error of the mean.