title: Heparan sulfate and heparin enhance ERK phosphorylation and mediate preBCR-dependent events during B lymphopoiesis.
abstract: As B lineage cells develop, they interact with cells, proteins, and extracellular matrix components of the surrounding microenvironment. In vitro, one critical checkpoint for developing cells occurs as they lose responsiveness to IL-7. These cells require contact with either stromal cells or other B lineage cells to mature. Our results demonstrate that heparan sulfate and heparin are able to promote this transition when added exogenously to the culture system or when heparan sulfate-bearing cell lines are cocultured with primary B cell progenitors. Addition of heparan sulfate or heparin to LPS-stimulated cultures of primary B cell progenitors resulted in more IgM secreted compared with untreated cultures. Heparan sulfate has been reported to be a ligand for the pre-B cell receptor (preBCR). Extending this observation, we found that treatment of preBCR+ cells with heparan sulfate before anti-micro stimulation leads to increased phosphorylation of ERK1/2. Consequently, preBCR+ cells proliferate more in the presence of IL-7 and heparan sulfate, whereas preBCR- cells are unaffected, suggesting that in these experiments, heparan sulfate is not directly affecting IL-7 activity. Heparin treatment of cultures induces many of the same biological effects as treatment with heparan sulfate, including elevated pERK levels in preBCR+ cells. However, heparin reduces the proliferation of cells expressing only the preBCR (opposed to both the preBCR and BCR) possibly due to internalization of the preBCR. Heparan sulfates are present on stromal cells and B lineage cells present in hemopoietic tissues and may provide stimulation to preB cells testing the signaling capacity of the preBCR.
authors.affiliation: Ontario Cancer Institute, University Health Network, Department of Immunology, University of Toronto, and Princess Margaret Hospital, Toronto, Ontario, Canada. cdmilne@ualberta.ca
authors.forename: Craig D
authors.initials: CD
authors.lastname: Milne
authors.forename: Steven A
authors.initials: SA
authors.lastname: Corfe
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Paige
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D002417:Cattle; D002450:Cell Communication / Q000276:immunology; D002454:Cell Differentiation / Q000276:immunology; D002460:Cell Line; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D006493:Heparin / Q000502:physiology*; D006497:Heparan Sulfate / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D007075:Immunoglobulin M / Q000096:biosynthesis; D038041:Lymphopoiesis / Q000276:immunology*; D051379:Mice; D016513:Mice, SCID; D008822:Mice, Transgenic; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D010766:Phosphorylation; D054420:Pre-B Cell Receptors / Q000378:metabolism / Q000502:physiology*; D013234:Stem Cells / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D013552:Swine
chemical_list: D006493:Heparin; D006497:Heparan Sulfate; D007075:Immunoglobulin M; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D054420:Pre-B Cell Receptors