Figure 4. AngII infusion causes a CC-chemokine dependent recruitment of monocytes to the aorta that is inhibited by R89A 35 K-Fc.

Mice were treated with AngII at 1 mg/kg/day by subcutaneous osmotic minipump. Aorta were harvested from mice following 3 days of infusion and RNA was prepared. A significant induction of CCL2 was measured in the mice following AngII infusion compared to baseline uninfused control animals (n = 4) (A). Digestion of isolated aortas showed CCR2+/CD11b+ leukocytes within the aorta (B) these cells were significantly increased in number following 7 days of Ang II infusion, vs saline control animals (n = 4–8) (C). Aortas harvested over a timecourse of AngII infusion showed a progressive recruitment of leuckoytes and in particular monocytes and macrophages to the aorta (n = 5–8) (D). To assess the efficacy of R89A 35K-Fc in inhibiting this recruitment, mice were injected with 1 μg R89A 35 K-Fc plasmid or GFP control plasmid 5 days prior to implanatation of an AngII minipump. Mice were sacrificed after 72 hours of AngII treatment (E). Analysis of plasma confirmed expression of 35 K-Fc by ELISA (F). Analysis of the recruited cells showed a significant blunting of the recruitment of total CD45+ leukocytes, CD11b+ myeloid cells and monocytes (inflammatory 7/4^HI and total 7/4^HI/INT) to the aorta, but no significant effect on neutrophil recruitment (G) (n = 12–13 AngII, n = 4 Saline). *p < 0.05. Statistical testing was performed by T-test (two groups only) or One-way ANOVA with Dunnett’s post testing.