title: Role of p38 MAPK and NF-kB for chemokine release in coculture of human eosinophils and bronchial epithelial cells.
abstract: Eosinophils are principal effector cells of inflammation in allergic asthma, characterized by their accumulation and infiltration at inflammatory sites mediated by the chemokine eotaxin and their interaction with adhesion molecules expressed on bronchial epithelial cells. We investigated the modulation of nuclear factor-kappaB (NF-kappaB) and the mitogen-activated protein kinase (MAPK) pathway on the in vitro release of chemokines including regulated upon activation normal T cell expressed and secreted (RANTES), monokine induced by interferon-gamma (MIG), monocyte chemoattractant protein-1 (MCP-1), interleukin (IL)-8, and interferon-inducible protein-10 (IP-10) upon the interaction of human bronchial epithelial BEAS-2B cells and eosinophils. Gene expression of chemokines was evaluated by RT-PCR and the induction amount of chemokines quantified by cytometric bead array. NF-kappaB and p38 MAPK activities were assessed by electrophoretic mobility shift assay and Western blot, respectively. The interaction of eosinophils and BEAS-2B cells was found to up-regulate the gene expression of the chemokines IL-8, MCP-1, MIG, RANTES and IP-10 expression in BEAS-2B cells, and to significantly elevate the release of the aforementioned chemokines except RANTES in a coculture of BEAS-2B cells and eosinophils. IkappaB-alpha phosphorylation inhibitor, BAY 11-7082, and p38 MAPK inhibitor, SB 203580 could decrease the release of IL-8, IP-10 and MCP-1 in the coculture. Together, the above results show that the induction of the release of chemokines in a coculture of epithelial cells and eosinophils are regulated by p38 MAPK and NF-kappaB activities of BEAS-2B cells, at least partly, through intercellular contact. Our findings therefore shed light on the future development of more effective agents for allergic and inflammatory diseases.
authors.affiliation: Department of Chemical Pathology, The Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, NT, Hong Kong.
authors.forename: C K
authors.initials: CK
authors.lastname: Wong
authors.forename: C B
authors.initials: CB
authors.lastname: Wang
authors.forename: W K
authors.initials: WK
authors.lastname: Ip
authors.forename: Y P
authors.initials: YP
authors.lastname: Tian
authors.forename: C W K
authors.initials: CW
authors.lastname: Lam
mesh_terms: D001980:Bronchi / Q000276:immunology*; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000276:immunology; D018946:Chemokine CCL5 / Q000276:immunology; D054357:Chemokine CXCL10; D054370:Chemokine CXCL9; D018925:Chemokines / Q000276:immunology*; D019743:Chemokines, CXC / Q000276:immunology; D018920:Coculture Techniques / Q000379:methods; D004791:Enzyme Inhibitors / Q000276:immunology; D004804:Eosinophils / Q000276:immunology*; D004847:Epithelial Cells / Q000276:immunology; D005404:Fixatives; D005557:Formaldehyde; D005786:Gene Expression Regulation / Q000276:immunology; D006801:Humans; D007093:Imidazoles / Q000276:immunology; D036341:Intercellular Signaling Peptides and Proteins / Q000276:immunology; D016209:Interleukin-8 / Q000276:immunology; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000276:immunology*; D009570:Nitriles / Q000276:immunology; D011108:Polymers; D011725:Pyridines / Q000276:immunology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013450:Sulfones / Q000276:immunology; D048051:p38 Mitogen-Activated Protein Kinases / Q000276:immunology*
chemical_list: C434003:3-(4-methylphenylsulfonyl)-2-propenenitrile; C111835:CCL2 protein, human; C114753:CXCL10 protein, human; C080810:CXCL9 protein, human; D018932:Chemokine CCL2; D018946:Chemokine CCL5; D054357:Chemokine CXCL10; D054370:Chemokine CXCL9; D018925:Chemokines; D019743:Chemokines, CXC; D004791:Enzyme Inhibitors; D005404:Fixatives; D007093:Imidazoles; D036341:Intercellular Signaling Peptides and Proteins; D016209:Interleukin-8; D016328:NF-kappa B; D009570:Nitriles; D011108:Polymers; D011725:Pyridines; D013450:Sulfones; D005557:Formaldehyde; D048051:p38 Mitogen-Activated Protein Kinases; C093642:SB 203580; C003043:paraform