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Stem Cells, Vol. 18, No. 2, 128-138, March 2000
© 2000 AlphaMed Press

The Role of HIV-Related Chemokine Receptors and Chemokines in Human Erythropoiesis in Vitro

Marcin Majkaa, Janina Ratajczaka, Benhur Leeb, Marek Honczarenkob, Ray Douglasb, M. Anna Kowalskac, Leslie Silbersteinb, Alan M. Gewirtza, Mariusz Z. Ratajczaka

a Department of Pathology & Laboratory Medicine;
b Department of Internal Medicine, Division of Hematology/Oncology, University of Pennsylvania School of Medicine;
c Department of Pediatrics, Children's Hospital of Philadelphia, Philadelphia, Pennsylvania, USA

Key Words. Hematopoiesis • Chemokines • Chemokine receptors • CXCR4 • CCR5 • Stromal derived factor-1 (SDF-1) • HIV

Mariusz Z. Ratajczak, M.D., Ph.D., Department of Pathology and Laboratory Medicine, University of Pennsylvania, 405A Stellar Chance Labs, 422 Curie Boulevard, Philadelphia, Pennsylvania 19104, USA. e-mail: mariusz{at}mail.med.upenn.edu

In order to better define the role of HIV-related chemokines in human erythropoiesis we studied: A) the expression of chemokine receptors, both on human CD34+ cells which include erythroid progenitors and on more mature erythroid cells; B) the functionality of these receptors by calcium flux, chemotaxis assay and phosphorylation of mitogen-activated protein kinases (MAPK) p42/44 (ERK1/ERK2) and AKT, and finally C) the influence of chemokines on BFU-E formation. We found that HIV-related chemokine receptor CXCR4, but not CCR5, is detectable on human CD34+ BFU-E cells. CXCR4 surface expression decreased during erythroid maturation, although CXCR4 mRNA was still present in cells isolated from differentiated erythroid colonies. SDF-1, a CXCR4 ligand, induced calcium flux and phosphorylation of MAPK (p42/44) and AKT in CD34+KIT+ bone marrow mononuclear cells which contain BFU-E, as well as chemotactic activity of both human CD34+ BFU-E progenitors and erythroid cells isolated from day 2-6 BFU-E colonies. Responsiveness to SDF-1 decreased when the cells differentiated to the point of surface expression of the erythroid-specific marker Glycophorin-A. In contrast, the CCR5 ligands (macrophage inflammatory protein-1{alpha} [MIP-1{alpha}], MIP-1ß, and RANTES) did not activate calcium flux, MAPK and AKT phosphorylation or chemotaxis of CD34+KIT+ cells or cells isolated from the BFU-E colonies. Interestingly, none of the chemokines tested in this study had any effect on BFU-E colony formation. In conclusion, only CXCR4 is functional, and its specific ligand SDF-1 may therefore play an important role in the homing and/or retention of early erythroid precursors in the bone marrow environment.




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