title: CXCR4 chemokine receptors (CD184) and alpha4beta1 integrins mediate spontaneous migration of human CD34+ progenitors and acute myeloid leukaemia cells beneath marrow stromal cells (pseudoemperipolesis).
abstract: Marrow stromal cells play an important role in regulating the development and proliferation of haematopoietic stem cells (HSC) within the marrow microenvironment. However, the molecular mechanisms of stem cell-stromal cell interactions are not fully understood. We observed that mobilized peripheral blood and cord-blood-derived CD34+ progenitor cells, or CD34+ acute myeloid leukaemia (AML) cells spontaneously migrated beneath marrow stromal cells, an in vitro migration phenomenon termed pseudoemperipolesis. In contrast, the CD34+ myeloid leukaemia cell line, Kasumi-1, did not display pseudoemperipolesis. Cord blood CD34+ cells had a higher capacity than granulocyte-colony-stimulating-factor-mobilized CD34+ cells for pseudoemperipolesis (28.7 +/- 12%vs 18.1 +/- 6.1% of input cells within 24 h, mean +/- SD, n = 8), whereas 9.4 +/- 12.6% (mean +/- SD, n = 10) of input AML cells displayed this phenomenon. Pseudoemperipolesis of CD34+ progenitor and AML cells was significantly inhibited by pertussis toxin and antibodies to the CXCR4 chemokine receptor (CXCR4, CD184), but not control antibodies. Moreover, CD34+ and AML cell migration was significantly inhibited by a CS1 peptide that blocks alpha4beta1 integrin binding, but not by a control peptide, in which the fibronectin binding motif was scrambled. Pseudoemperipolesis was associated with an increased proliferation of migrated CD34+ progenitor cells but not AML cells within the stromal layer, demonstrated by cell cycle analysis and cell division tracking. We conclude that alpha4beta1 integrin binding and CXCR4 chemokine receptor activation are prerequisites for the migration of CD34+ haematopoietic progenitors and AML cells beneath marrow stromal cells. These observations suggest a central role of marrow stromal cells for HSC trafficking and homing within the marrow microenvironment.
authors.affiliation: Division of Haematology/Oncology, Department of Medicine, Freiburg University Hospital, Hugstetterstrasse 55, D-79106 Freiburg, Germany. burger@mm11.ukl.uni-freiburg.de
authors.forename: Jan A
authors.initials: JA
authors.lastname: Burger
authors.forename: Anke
authors.initials: A
authors.lastname: Spoo
authors.forename: Anne
authors.initials: A
authors.lastname: Dwenger
authors.forename: Meike
authors.initials: M
authors.lastname: Burger
authors.forename: Dirk
authors.initials: D
authors.lastname: Behringer
mesh_terms: D018952:Antigens, CD34 / Q000032:analysis; D001854:Bone Marrow Cells / Q000502:physiology*; D002453:Cell Cycle / Q000502:physiology; D002455:Cell Division / Q000502:physiology; D002465:Cell Movement; D018920:Coculture Techniques; D005434:Flow Cytometry / Q000379:methods; D006412:Hematopoietic Stem Cells / Q000502:physiology*; D006801:Humans; D039041:Integrin alpha4beta1 / Q000502:physiology*; D007951:Leukemia, Myeloid / Q000473:pathology*; D008024:Ligands; D019718:Receptors, CXCR4 / Q000502:physiology*; D017154:Stromal Cells / Q000502:physiology; D014407:Tumor Cells, Cultured
chemical_list: D018952:Antigens, CD34; D039041:Integrin alpha4beta1; D008024:Ligands; D019718:Receptors, CXCR4