Abstract 372: CD16 Monocytes Stimulate Arteriogenesis to Salvage the Ischemic Limb

Introduction: Cell therapy using unselected mononuclear cell populations has had modest benefits in patients with critical limb ischemia (CLI). We hypothesized that tissue-remodeling monocytes, identified by their expression of CD16 (CD16+ Mo), may be a novel cellular therapy for CLI. Methods and Results: Flow cytometry showed that the proportion of circulating CD16+ Mo was greater in CLI patients (n=25) compared with matched controls (n=15, P<0.0001). Removal of ischemia following revascularization or amputation resulted in a fall in CD16+ Mo to control levels (P<0.05). CLI CD16+ Mo expressed higher levels of the adhesive proteins VLA, ICAM-1 and CD11c compared with controls (P<0.05). Conditioned media from these cells contained higher levels of HB-EGF, PlGF, endoglin, VEGF-C and VEGF-D (P<0.05) and induced greater endothelial cell tubule formation (P<0.05) compared with CD16- Mo from the same patients (n=9). CD16+ Mo preferentially migrated towards ischemic muscle supernatants isolated from CLI patients (n=7, P<0.02). CD16+ and CD16- Mo were isolated from 12 CLI patients and 1x10 6 cells injected into the adductor muscles of nude athymic mice following femoral artery excision. More ischemic hindlimbs were salvaged when treated with CD16+ compared with CD16- Mo (83% [10/12] vs 17% [2/12] limbs, P<0.05) and this was associated with enhanced arteriogenesis (αSMA-stained vessels, P<0.05). Conclusion: Circulating CD16+ Mo from CLI patients have increased expression of adhesion markers, are preferentially retained within ischemic muscle and promote robust arteriogenesis and limb salvage in experimental HLI. This monocyte subset may be an effective cellular therapy for CLI.