CD44 mediates hyaluronan binding by human myeloid KG1A and KG1 cells.
Hyaluronan-binding function of the CD44 molecule has not been so far detected in myeloid cells. To study pure populations of primitive myeloid cells, we investigated the hyaluronan-binding function of the CD44 molecule from three myeloid cell lines: KG1a, KG1, and HL60. Both KG1a and KG1 cells express the CD34 antigen characteristic of the hematopoietic stem cells and HL60 cells do not; accordingly, KG1a and KG1 cells are generally considered as the most primitive and HL60 cells as the most mature of these cell lines. Measurement of cell adhesion to hyaluronan-coated surfaces (using 51Cr-labeled cells) and of aggregate formation in hyaluronan-containing solutions, showed that 45% of KG1 cells and 22% to 24% of KG1a spontaneously bind to hyaluronan, whereas HL60 cells do not either spontaneously or after treatment with a phorbol ester. Hyaluronan binding by KG1a and KG1 cells is mediated by CD44, because it is specifically abolished by monoclonal antibodies (MoAbs) to this molecule. The binding might require phosphorylation by protein kinase C and perhaps also by protein kinase A, because it is prevented by staurosporine, which inhibits these enzymes. 12-O-tetradecanoylphorbol-13-acetate (TPA) which activates protein kinase C, rises to 80% the proportion of KG1 and KG1a cells that bind hyaluronan; this activation is dependent on protein synthesis, for it is abrogated by cyclophosphamide, a protein synthesis inhibitor. Binding of TPA-treated cells to hyaluronan is only partly inhibited by MoAb to CD44: this suggests that TPA may induce synthesis of a hyaluronan-binding protein distinct from CD44. Considering the abundance of hyaluronan in human bone marrow, these results suggest that CD44 may be involved in mediating precursor-stroma interaction.
Morimoto
K
K
Unité d'Oncogénèse Appliquée, Inserm U268, Hopital Paul Brousse, Villejuif, France.
Robin
E
E
Le Bousse-Kerdiles
M C
MC
Li
Y
Y
Clay
D
D
Jasmin
C
C
Smadja-Joffe
F
F
D000470:Alkaloids / Q000494:pharmacology; D001853:Bone Marrow / Q000378:metabolism; D001854:Bone Marrow Cells; D002352:Carrier Proteins / Q000032:analysis / Q000502:physiology*; D002448:Cell Adhesion; D002449:Cell Aggregation; D002460:Cell Line; D006801:Humans; D018960:Hyaluronan Receptors; D006820:Hyaluronic Acid / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000032:analysis / Q000502:physiology*; D016201:Receptors, Lymphocyte Homing / Q000032:analysis / Q000502:physiology*; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D000470:Alkaloids; D002352:Carrier Proteins; D018960:Hyaluronan Receptors; D011956:Receptors, Cell Surface; D016201:Receptors, Lymphocyte Homing; D006820:Hyaluronic Acid; D019311:Staurosporine; D013755:Tetradecanoylphorbol Acetate