title: IL-35 Suppresses Lipopolysaccharide-Induced Airway Eosinophilia in EBI3-Deficient Mice.
abstract: EBI3 functions as the subunit of immune-regulatory cytokines, such as IL-27 and IL-35, by pairing with p28 and p35, respectively. We treated wild-type and EBI3-deficient mice with intratracheal administration of LPS and obtained bronchoalveolar lavage fluid (BALF) 24 h later. Although neutrophils were the predominant cells in BALF from both groups of mice, eosinophils were highly enriched and there was increased production of eosinophil-attracting chemokines CCL11 and CCL24 in BALF of EBI3-deficient mice. The bronchial epithelial cells and alveolar macrophages were the major producers of CCL11 and CCL24. Because no such increases in eosinophils were seen in BALF of p28/IL-27-deficient mice or WSX-1/IL-27Rα subunit-deficient mice upon intratracheal stimulation with LPS, we considered that the lack of IL-35 was responsible for the enhanced airway eosinophilia in EBI3-deficient mice. In vitro, IL-35 potently suppressed production of CCL11 and CCL24 by human lung epithelial cell lines treated with TNF-α and IL-1β. IL-35 also suppressed phosphorylation of STAT1 and STAT3 and induced suppressor of cytokine signaling 3. In vivo, rIL-35 dramatically reduced LPS-induced airway eosinophilia in EBI3-deficient mice, with concomitant reduction of CCL11 and CCL24, whereas neutralization of IL-35 significantly increased airway eosinophils in LPS-treated wild-type mice. Collectively, our results suggest that IL-35 negatively regulates airway eosinophilia, at least in part by reducing the production of CCL11 and CCL24.
authors.affiliation: Department of Microbiology, Kindai University Faculty of Medicine, Osaka 589-8511, Japan; and.
authors.forename: Kyosuke
authors.initials: K
authors.lastname: Kanai
authors.affiliation: Department of Microbiology, Kindai University Faculty of Medicine, Osaka 589-8511, Japan; and.
authors.forename: Ah-Mee
authors.initials: AM
authors.lastname: Park
authors.affiliation: Division of Molecular and Cellular Immunoscience, Department of Biomolecular Sciences, Saga University Faculty of Medicine, Saga 840-8502, Japan.
authors.forename: Hiroki
authors.initials: H
authors.lastname: Yoshida
authors.affiliation: Department of Microbiology, Kindai University Faculty of Medicine, Osaka 589-8511, Japan; and.
authors.forename: Ikuo
authors.identifier: 0000-0003-1798-714X
authors.initials: I
authors.lastname: Tsunoda
authors.affiliation: Department of Microbiology, Kindai University Faculty of Medicine, Osaka 589-8511, Japan; and qqud7p89k@siren.ocn.ne.jp.
authors.forename: Osamu
authors.initials: O
authors.lastname: Yoshie
mesh_terms: D000818:Animals; D002460:Cell Line; D054413:Chemokine CCL11 / Q000096:biosynthesis; D054423:Chemokine CCL24 / Q000096:biosynthesis; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D007378:Interleukins / Q000276:immunology* / Q000378:metabolism; D008070:Lipopolysaccharides / Q000276:immunology / Q000633:toxicity; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D015778:Minor Histocompatibility Antigens / Q000276:immunology*; D016133:Polymerase Chain Reaction; D011657:Pulmonary Eosinophilia / Q000276:immunology* / Q000473:pathology; D018121:Receptors, Cytokine / Q000172:deficiency / Q000276:immunology*
chemical_list: D054413:Chemokine CCL11; D054423:Chemokine CCL24; C499601:Ebi3 protein, mouse; D007378:Interleukins; D008070:Lipopolysaccharides; D015778:Minor Histocompatibility Antigens; D018121:Receptors, Cytokine; C569375:interleukin-35, mouse