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Expression of LTC4 synthase during the development of eosinophils in vitro
from cord blood progenitors
JA Boyce, BK Lam, JF Penrose, DS Friend, S Parsons, WF Owen and KF Austen
Department of Medicine, Harvard Medical School, Boston, MA, USA.
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.
Volume 88,
Issue 11,
pp. 4338-4347,
12/01/1996
Copyright © 1996 by The American Society of Hematology
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