T helper cell type 2 cytokines coordinately regulate immunoglobulin E-dependent cysteinyl leukotriene production by human cord blood-derived mast cells: profound induction of leukotriene C(4) synthase expression by interleukin 4.
Human mast cells (hMCs) derived in vitro from cord blood mononuclear cells exhibit stem cell factor (SCF)-dependent comitogenic responses to T helper cell type 2 (Th2) cytokines. As cysteinyl leukotriene (cys-LT) biosynthesis is a characteristic of immunoglobulin (Ig)E-activated mucosal hMCs, we speculated that Th2 cytokines might regulate eicosanoid generation by hMCs. After passive sensitization for 5 d with IgE in the presence of SCF, anti-IgE-stimulated hMCs elaborated minimal cys-LT (0.1 +/- 0.1 ng/10(6) hMCs) and abundant prostaglandin (PG)D(2) (16.2 +/- 10.3 ng/10(6) hMCs). Priming of hMCs by interleukin (IL)-4 with SCF during passive sensitization enhanced their anti-IgE-dependent histamine exocytosis and increased their generation of both cys-LT (by 27-fold) and PGD(2) (by 2. 5-fold). Although priming with IL-3 or IL-5 alone for 5 d with SCF minimally enhanced anti-IgE-mediated cys-LT generation, these cytokines induced further six- and fourfold increases, respectively, in IgE-dependent cys-LT generation when provided with IL-4 and SCF; this occurred without changes in PGD(2) generation or histamine exocytosis relative to hMCs primed with IL-4 alone. None of these cytokines, either alone or in combination, substantially altered the levels of cytosolic phospholipase A(2) (cPLA(2)), 5-lipoxygenase (5-LO), or 5-LO activating protein (FLAP) protein expression by hMCs. In contrast, IL-4 priming dramatically induced the steady-state expression of leukotriene C(4) synthase (LTC(4)S) mRNA within 6 h, and increased the expression of LTC(4)S protein and functional activity in a dose- and time-dependent manner, with plateaus at 10 ng/ml and 5 d, respectively. Priming by either IL-3 or IL-5, with or without IL-4, supported the localization of 5-LO to the nucleus of hMCs. Thus, different Th2-derived cytokines target distinct steps in the 5-LO/LTC(4)S biosynthetic pathway (induction of LTC(4)S expression and nuclear import of 5-LO, respectively), each of which is necessary for a full integrated functional response to IgE-dependent activation, thus modulating the effector phenotype of mature hMCs.
Hsieh
F H
FH
Department of Medicine, Harvard Medical School, Boston, Massachusetts 02115, USA.
Lam
B K
BK
Penrose
J F
JF
Austen
K F
KF
Boyce
J A
JA
D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism; D002467:Cell Nucleus / Q000201:enzymology; D003545:Cysteine / Q000096:biosynthesis*; D016207:Cytokines / Q000494:pharmacology* / Q000502:physiology; D005312:Fetal Blood / Q000166:cytology / Q000276:immunology; D005982:Glutathione Transferase / Q000096:biosynthesis*; D006801:Humans; D007073:Immunoglobulin E / Q000378:metabolism*; D066298:In Vitro Techniques; D007231:Infant, Newborn; D007377:Interleukin-3 / Q000494:pharmacology; D015848:Interleukin-5 / Q000494:pharmacology; D015289:Leukotrienes / Q000096:biosynthesis*; D008407:Mast Cells / Q000187:drug effects / Q000201:enzymology / Q000276:immunology*; D015230:Prostaglandin D2 / Q000096:biosynthesis; D019089:Stem Cell Factor / Q000494:pharmacology; D018418:Th2 Cells / Q000276:immunology*
D016207:Cytokines; D007377:Interleukin-3; D015848:Interleukin-5; D015289:Leukotrienes; D019089:Stem Cell Factor; C112381:cysteinyl-leukotriene; D007073:Immunoglobulin E; D001094:Arachidonate 5-Lipoxygenase; D005982:Glutathione Transferase; C041116:leukotriene-C4 synthase; D003545:Cysteine; D015230:Prostaglandin D2