SOD3 Derived Cell Migration Figure 1. Reduced macrophage infiltration in to ischemic muscle. Open bars represent LacZ animals and black bars represent SOD3 animals. CD68 staining showed significantly reduced inflammatory area and macrophage infiltration in SOD3 animals at all time points studied. Histological stainings show CD68+ macrophages around the femoral artery in ischemic muscles (206 magnification). doi:10.1371/journal.pone.0005786.g001 regulating the inflammatory cell migration into the injuries. In the present work we studied leukocyte migration in acute ischemia and peritonitis models and analyzed the contribution of SOD3 on inflammatory cytokine and adhesion molecule expression. We determined the effect of SOD3 overexpression on the degree of inflammation by analyzing the size of inflamed tissue and the number of infiltrated macrophages and T cells in acute ischemic injury model. In a mouse model of femoral artery ligation, macrophage infiltration into ischemic muscle reaches peak values 3 days after the injury [39]. Histological analysis of the rat hind limb ischemia showed 3-fold reduction in the inflamed tissue area as determined by the presence of CD68+ macrophages (p,0.001) in SOD3 vs. LacZ control animals 3 days after vessel ligation (Figure 1). The reduction became even more prominent in later time points reaching 12-fold difference 10 days after vessel ligation. Additionally, the number of infiltrated CD68+ macro- phages was 3–5 fold higher in LacZ control animals as compared to SOD3 animals (p,0.05). Maximal macrophage accumulation to the injured tissue was seen at 7-day time point in LacZ animals indicating that the inflammatory reaction was still developing in control animals at the initial phase of the follow-up period while the inflammation was decreasing in SOD3 animals. Throughout the experiment the number of macrophages remained higher in control animals than initially observed in SOD3 animals, which by the 10-day time point showed values close to the background levels further underlining the beneficial effect of SOD3. PLoS ONE | www.plosone.org 3 June 2009 | Volume 4 | Issue 6 | e5786