title: 1 alpha,25-Dihydroxyvitamin D3 promotes monocytopoiesis and suppresses granulocytopoiesis in cultures of normal human myeloid blast cells.
abstract: Primitive myeloid blast cells (2-10 x 10(6)) were purified from 18-22-week fetal liver-derived mononuclear cell preparations by negative selection followed by counterflow cell elutriation. The cells, when maintained in liquid culture in the presence of 100 U/ml interleukin-3 (IL-3) for the first 5 days and 10 U/ml IL-3 and 30 ng/ml granulocyte colony-stimulating factor thereafter, underwent considerable proliferation resulting in an approximately 30-fold increase in cell number by day 14. Analyses of cell morphology and of the numbers of cells that expressed the neutrophil-associated antigen CD15, the monocyte-associated antigen 61D3, and enzymes alpha-naphthyl acetate esterase (ANAE), human leukocyte elastase, and cathepsin G revealed that proliferation of the cells was associated with their concomitant differentiation toward neutrophils and monocytes. The cultures generated predominantly neutrophils; by day 14, wells seeded with 2 x 10(5) cells produced approximately 5 x 10(6) neutrophils as opposed to only approximately 3.5 x 10(5) cells with a monocytoid morphology. This predominance of granulocytopoiesis over monocytopoiesis was confirmed by the numbers of cells that had acquired expression of the CD15 antigen and ANAE, which were approximately 2 x 10(6) and 1 x 10(5), respectively. By contrast, parallel cultures containing 100 nM 1 alpha,25-dihydroxyvitamin D3 (VitD3) generated more monocytes than neutrophils. At day 14, VitD3-treated cultures contained approximately 2 x 10(6) cells with morphologies consistent with their differentiation toward monocytes and approximately 1 x 10(6) ANAE-positive cells, compared with approximately 9.5 x 10(5) cells having morphologies of granulocyte-series cells and approximately 4.5 x 10(4) CD15-positive cells. In both control and VitD3-treated cultures, the enzymes cathepsin G and human leukocyte elastase were expressed almost exclusively by cells that were differentiating toward neutrophils. These data reveal that VitD3 promotes monocytopoiesis and suppresses granulocytopoiesis of primitive blast cells.
authors.affiliation: Department of Immunology, Medical School, University of Birmingham, England.
authors.forename: A E
authors.initials: AE
authors.lastname: Barton
authors.forename: C M
authors.initials: CM
authors.lastname: Bunce
authors.forename: R A
authors.initials: RA
authors.lastname: Stockley
authors.forename: P
authors.initials: P
authors.lastname: Harrison
authors.forename: G
authors.initials: G
authors.lastname: Brown
mesh_terms: D002117:Calcitriol / Q000494:pharmacology*; D056649:Cathepsin G; D002403:Cathepsins / Q000502:physiology; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology; D002454:Cell Differentiation / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D006098:Granulocytes / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D006410:Hematopoiesis / Q000187:drug effects*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects; D006801:Humans; D019272:Leukocyte Elastase; D008213:Lymphocyte Activation / Q000187:drug effects; D008954:Models, Biological; D009000:Monocytes / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology; D010196:Pancreatic Elastase / Q000502:physiology; D010586:Phagocytes / Q000187:drug effects; D012697:Serine Endopeptidases
chemical_list: D002403:Cathepsins; D012697:Serine Endopeptidases; C534094:CTSG protein, human; D056649:Cathepsin G; D010196:Pancreatic Elastase; D019272:Leukocyte Elastase; D002117:Calcitriol