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a Division of Medical Oncology, Netherlands Cancer Institute, Amsterdam, The Netherlands;
b CLB, Sanquin Blood Supply Foundation, and Laboratory for Experimental and Clinical Immunology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands;
c Gene Therapy Program, Department of Medical Oncology, Academic Hospital of the Free University, Amsterdam, The Netherlands;
d Department of Hematology, Academic Medical Center, Amsterdam, The Netherlands
Key Words. Migration • SDF-1 • Hematopoietic progenitor cells • Bone marrow endothelial cells • Adhesion molecules • Homing
C.E. van der Schoot, M.D., Ph.D., Department of Immunohematology, Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Plesmanlaan 125, 1066 CX Amsterdam, The Netherlands. Telephone: 31-20-5123377; Fax: 31-20-5123474; e-mail: Schoot{at}clb.nl
In the process of homing, CD34+ hematopoietic progenitor cells migrate across the bone marrow endothelium in response to stromal cell-derived factor (SDF)-1. To develop more efficient stem cell transplantation procedures, it is important to define the adhesion molecules involved in the homing process. Here, we identified the adhesion molecules that control the migration of primary human CD34+ cells across human bone marrow endothelial cells.
Migration of CD34+ cells is enhanced across interleukin 1ß prestimulated bone marrow endothelium, suggesting an important role for the endothelium in adhesion and formation of the chemotactic gradient. Under these conditions, 30-100 ng/ml SDF-1 induced a rapid and efficient migration of CD34+ cells (± 46% migration in 4 h). In contrast, 600-1,000 ng/ml SDF-1 were required for optimal migration across fibronectin-coated filters. Subsequent studies revealed that transendothelial migration of CD34+ cells is mediated by ß1- and ß2-integrins and PECAM-1 (CD31) but not by CD34 or E-selectin. Whereas these antibodies individually blocked migration for 25%-35%, migration was reduced by 68% when the antibodies were combined. Thus, these adhesion molecules play specific and independent roles in the transmigration process. Finally, O-glycosylated proteins appeared to play a role, since SDF-1-induced migration of CD34+ cells (treated with a glycoprotease from Pasteurella haemolytica) across endothelial cells was clearly inhibited.
In conclusion, we show that efficient SDF-1-induced migration of primary human CD34+ cells across bone marrow endothelium is mediated by ß1-integrins, ß2-integrins, CD31 and O-glycosylated proteins.
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