Interleukin-3 regulates development of the 5-lipoxygenase/leukotriene C4 synthase pathway in mouse mast cells.
To study cytokine regulation of the 5-lipoxygenase (5-LO)/leukotriene (LT) synthase pathway we have developed mouse bone marrow-derived mast cells (BMMC) that minimally express each protein of the pathway by using a novel culture system, lacking interleukin (IL)-3. When mouse bone marrow cells were cultured for 5 weeks with 100 ng/ml c-kit ligand (KL) and 10 units/ml IL-10, a population of > 95% mast cells was obtained. These cells generated 8.3 +/- 4.5 ng of LTC4/10(6) cells and 8.1 +/- 2.4 ng of prostaglandin (PG) D2/10(6) cells after IgE-dependent activation. When these BMMC were cultured for 2-5 weeks more with 100 units/ml IL-3 in the continued presence of KL and IL-10, the IgE-dependent generation of LTC4 and PGD2 increased to 212 +/- 36 and 25.5 +/- 8.6 ng/10(6) cells, respectively. The dramatic increase in the IgE-dependent generation of LTC4 in response to IL-3 was accompanied by a concomitant increase in expression of 5-LO and 5-LO-activating protein and preceded the increased expression of cytosolic phospholipase A2 and LTC4 synthase. The recognition that IL-3 up-regulates the expression of each protein of the 5-LO pathway for the generation of LTC4 contrasts with our recent finding that KL up-regulates the expression of cytosolic phospholipase A2, prostaglandin endoperoxide synthase-1, and hematopoietic PGD2 synthase and increases the IgE-dependent generation of PGD2 in BMMC developed from bone marrow with IL-3. Thus, developmentally segregated regulation of the prostanoid and cysteinyl leukotriene pathways in lineage-related committed mast cell progenitors reveals the pleiotropism of this effector cell of allergic inflammation, a cytokine/growth factor basis for preferential expression of pathways of eicosanoid biosynthesis, and the particular role of IL-3 in regulating the expression of the proteins of the 5-LO/LTC4 synthase pathway.
Murakami
M
M
Department of Medicine, Harvard Medical School, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA.
Austen
K F
KF
Bingham
C O
CO
Friend
D S
DS
Penrose
J F
JF
Arm
J P
JP
D000818:Animals; D001094:Arachidonate 5-Lipoxygenase / Q000096:biosynthesis*; D001854:Bone Marrow Cells; D002478:Cells, Cultured; D015870:Gene Expression / Q000187:drug effects*; D005982:Glutathione Transferase / Q000096:biosynthesis*; D006133:Growth Substances / Q000494:pharmacology*; D006412:Hematopoietic Stem Cells / Q000166:cytology; D006801:Humans; D007073:Immunoglobulin E / Q000494:pharmacology; D007377:Interleukin-3 / Q000494:pharmacology*; D007700:Kinetics; D017997:Leukotriene C4 / Q000096:biosynthesis; D008407:Mast Cells / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D051379:Mice; D010741:Phospholipases A / Q000096:biosynthesis; D054467:Phospholipases A2; D015230:Prostaglandin D2 / Q000494:pharmacology; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000494:pharmacology; D019089:Stem Cell Factor / Q000494:pharmacology; D013997:Time Factors
D006133:Growth Substances; D007377:Interleukin-3; D011994:Recombinant Proteins; D019089:Stem Cell Factor; D017997:Leukotriene C4; D007073:Immunoglobulin E; D001094:Arachidonate 5-Lipoxygenase; D011451:Prostaglandin-Endoperoxide Synthases; D005982:Glutathione Transferase; D010741:Phospholipases A; D054467:Phospholipases A2; C041116:leukotriene-C4 synthase; D015230:Prostaglandin D2