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Endothelial Progenitor Cells Are Mobilized In Critical Limb Ischemia

Julie C Mund, Dana L. Booth, MD, Shoaib Shafique, MD, Ryan D. Nachreiner, MD, Joseph L. Unthank, PhD, Michael C. Dalsing, MD, Michael P. Murphy, MD ∙ Indiana University School of Medicine, Indianapolis, IN

Objectives: There is little known as to the cellular processes which lead to the development of peripheral vascular disease. Circulating endothelial progenitor cells (EPC) are thought to play a role in the development and maintenance of vessel walls. These cells are identified by the cell surface markers CD34, CD133, and VEGF-R2. We report our preliminary findings comparing EPCs in patients with CLI and healthy donors.
Methods: We collected 100 mL samples of peripheral blood from patients with CLI, as determined by an ankle-brachial index of <0.5 and healthy donors (ABI >0.90). Bone marrow (50mL.) was also obtained from some donors in each group. After isolation, the mononuclear cell fraction was labeled with CD14, CD34, CD45, CD105, CD133, and VEGF-R2 antibodies for fluorescent activated cell sorting (FACS) analysis. Results: The patients with CLI had significantly down regulated expression of CD105 when compared to healthy donors both in the peripheral circulation (1.51 +/- 1.30 vs. 5.38 +/- 8.25) and in the bone marrow cells (3.27 +/- 2.26 vs. 7.95 +/- 9.07). The patients with CLI grew more early endothelial progenitor cell colonies than healthy donors (63.6 compared to 154.6 per 5X10^6 cells plated). This trend also persisted in the late outgrowth endothelial progenitor cell colonies with healthy donors growing 1.2 colonies and CLI patients 7.53 colonies (per 50X106 cells plated). Conclusion: These preliminary results indicate that CD105 is down regulated in patients with severe peripheral vascular disease, however the most novel find is that patients with CLI are still able to mobilize EPCs,contrary to earlier published reports by other investigators. More data is needed to confirm this finding and to test the functionality of the cells in both the healthy donors for comparison to the CLI patients.