title: Essential roles of PKA, iNOS, CD95/CD95L, and terminal caspases in suppression of eosinopoiesis by PGE2 and other cAMP-elevating agents.
abstract: Up- and downregulation of eosinopoiesis control pulmonary eosinophilia in human asthma. In mice, eosinopoiesis is suppressed in vitro by prostaglandin E2 (PGE2) and in vivo by diethylcarbamazine, through a proapoptotic mechanism sequentially requiring inducible NO synthase (iNOS) and the ligand for death receptor CD95 (CD95L). We examined the roles of iNOS, cAMP-mediated signaling, caspases, and CD95L/CD95 in suppression of eosinopoiesis by PGE2 and other agents signaling through cAMP. Bone-marrow collected from BALB/c mice, or from iNOS-, CD95-, or CD95L-deficient mutants (and wild-type controls), was cultured with interleukin-5 (IL-5), alone or associated with PGE2, cAMP-inducing/mimetic agents, caspase inhibitor zVAD-fmk, iNOS inhibitor aminoguanidine, or combinations thereof, and eosinopoiesis was evaluated at various times. PGE2, added up to 24 hours of culture, dose-dependently suppressed eosinopoiesis, by inducing apoptosis. This effect was (a) paralleled by induction of iNOS in eosinophils; (b) duplicated by sodium nitroprusside, isoproterenol, and cAMP-inducing/mimetic agents; (c) prevented by protein kinase A inhibition. NO was produced through iNOS by dibutyryl-cAMP-stimulated bone-marrow. Overall, PGE2 and isoproterenol shared a requirement for four effector elements (iNOS, CD95L, CD95, and terminal caspases), which together define a pathway targeted by several soluble up- and downmodulators of eosinopoiesis, including drugs, mediators of inflammation, and cytokines.
authors.affiliation: Department of Pediatrics, Instituto Nacional de Saúde da Mulher, da Criança e do Adolescente Fernandes Figueira, FIOCRUZ, Avenue Rui Barbosa 716, 22250-020 Rio de Janeiro, RJ, Brazil.
authors.forename: Bianca
authors.initials: B
authors.lastname: de Luca
authors.affiliation: Department of Immunology, Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, CCS, Bloco I, Room I-2-066, 22205-020, Rio de Janeiro, Brazil.
authors.forename: Pedro
authors.initials: P
authors.lastname: Xavier-Elsas
authors.affiliation: Department of Immunology, Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, CCS, Bloco I, Room I-2-066, 22205-020, Rio de Janeiro, Brazil.
authors.forename: Mônica
authors.initials: M
authors.lastname: Barradas
authors.affiliation: Department of Immunology, Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, CCS, Bloco I, Room I-2-066, 22205-020, Rio de Janeiro, Brazil.
authors.forename: Ricardo A
authors.initials: RA
authors.lastname: Luz
authors.affiliation: Department of Pediatrics, Instituto Nacional de Saúde da Mulher, da Criança e do Adolescente Fernandes Figueira, FIOCRUZ, Avenue Rui Barbosa 716, 22250-020 Rio de Janeiro, RJ, Brazil.
authors.forename: Túlio
authors.initials: T
authors.lastname: Queto
authors.affiliation: Department of Immunology, Instituto de Microbiologia Professor Paulo de Góes, Universidade Federal do Rio de Janeiro, CCS, Bloco I, Room I-2-066, 22205-020, Rio de Janeiro, Brazil.
authors.forename: Carla
authors.initials: C
authors.lastname: Jones
authors.affiliation: Farmanguinhos, FIOCRUZ, Avenue Comandante Guaranys No. 447, Jacarepaguá, 22775-903 Rio de Janeiro, RJ, Brazil.
authors.forename: Maria Augusta
authors.initials: MA
authors.lastname: Arruda
authors.affiliation: Department of Pharmacology, Faculdade de Medicina da USP, Avenue Bandeirantes 3900, Monte Alegre, 14049-900 Ribeirão Preto, SP, Brazil.
authors.forename: Thiago Mattar
authors.initials: TM
authors.lastname: Cunha
authors.affiliation: Department of Pharmacology, Faculdade de Medicina da USP, Avenue Bandeirantes 3900, Monte Alegre, 14049-900 Ribeirão Preto, SP, Brazil.
authors.forename: Fernando Queiroz
authors.initials: FQ
authors.lastname: Cunha
authors.affiliation: Department of Pediatrics, Instituto Nacional de Saúde da Mulher, da Criança e do Adolescente Fernandes Figueira, FIOCRUZ, Avenue Rui Barbosa 716, 22250-020 Rio de Janeiro, RJ, Brazil.
authors.forename: Maria Ignez
authors.initials: MI
authors.lastname: Gaspar-Elsas
mesh_terms: D000262:Adenylyl Cyclases / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells / Q000187:drug effects; D061945:Caspase Inhibitors / Q000494:pharmacology; D020169:Caspases / Q000378:metabolism*; D002478:Cells, Cultured; D000242:Cyclic AMP / Q000378:metabolism; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D003907:Dexamethasone / Q000494:pharmacology; D015232:Dinoprostone / Q000494:pharmacology*; D004804:Eosinophils / Q000378:metabolism*; D053222:Fas Ligand Protein / Q000235:genetics / Q000378:metabolism*; D015848:Interleukin-5 / Q000494:pharmacology; D007545:Isoproterenol / Q000494:pharmacology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008817:Mice, Mutant Strains; D052247:Nitric Oxide Synthase Type II / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D019014:fas Receptor / Q000235:genetics / Q000378:metabolism*
chemical_list: D061945:Caspase Inhibitors; D053222:Fas Ligand Protein; D015848:Interleukin-5; D019014:fas Receptor; D003907:Dexamethasone; D000242:Cyclic AMP; D052247:Nitric Oxide Synthase Type II; C496317:Nos2 protein, mouse; D017868:Cyclic AMP-Dependent Protein Kinases; D020169:Caspases; D000262:Adenylyl Cyclases; D015232:Dinoprostone; D007545:Isoproterenol