Expression of LTC4 synthase during the development of eosinophils in vitro from cord blood progenitors
| Autor: | J A, Boyce, B K, Lam, J F, Penrose, D S, Friend, S, Parsons, W F, Owen, K F, Austen |
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| Rok vydání: | 1996 |
| Předmět: |
Leukotrienes
5-Lipoxygenase-Activating Proteins Lipoxygenase Immunology Biochemistry Phospholipases A Humans RNA Messenger Calcimycin Cells Cultured Glutathione Transferase Ionophores Infant Newborn Membrane Proteins Cell Biology Hematology Fetal Blood Hematopoietic Stem Cells Recombinant Proteins Eosinophils Drug Combinations Phospholipases A2 Phenotype Enzyme Induction Calcium Interleukin-3 Proteoglycans Collagen Laminin Interleukin-5 Carrier Proteins |
| Zdroj: | Blood. 88:4338-4347 |
| ISSN: | 1528-0020 0006-4971 |
| DOI: | 10.1182/blood.v88.11.4338.bloodjournal88114338 |
| Popis: | The expression of leukotriene C4 synthase (LTC4S) was examined during the development of eosinophils in vitro from cord blood mononuclear cells. At 7 days, the cells contained mRNA and sodium dodecyl sulfate-polyacrylamide gel electrophoresis immunoblot signals for cytosolic phospholipase A2 (cPLA2), 5-lipoxygenase (5-LO), and 5-lipoxygenase-activating protein (FLAP), but lacked LTC4S and did not generate cysteinyl leukotrienes when stimulated with 20 mumol/L calcium ionophore. At 14 days, 94% of the cells were of eosinophil lineage, both LTC4S mRNA transcript and protein were present, and ionophore stimulation resulted in the generation of 23.9 +/- 6.0 pmol cysteinyl leukotrienes/10(6) eosinophil-lineage cells (mean +/- SEM, n = 6). At 28 days, progressive eosinophil maturation was accompanied by further increments in 5-LO, FLAP, and LTC4S proteins, and by the ionophore- induced production of 94.6 +/- 9.0 pmol cysteinyl leukotrienes/10(6) eosinophil-lineage cells (n = 6). Cells selected for CD34 expression lacked detectable 5-LO/LTC4S pathway proteins, and with culture generally expressed immunodetectable cPLA2 and 5-LO proteins by 3 days, FLAP protein by 7 days, and LTC4S protein by 10 days. Thus, during the development of eosinophils in vitro, cPLA2, 5-LO, and FLAP are expressed before LTC4S. Once the lineage is established by morphologic criteria, the eosinophilopoietic cytokines mediate upregulation of FLAP and LTC4S, members of a newly recognized gene family, and of 5-LO, during ongoing cell maturation. |
| Databáze: | OpenAIRE |
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