title: IL-33 markedly activates murine eosinophils by an NF-κB-dependent mechanism differentially dependent upon an IL-4-driven autoinflammatory loop.
abstract: Eosinophils are major effector cells in type 2 inflammatory responses and become activated in response to IL-4 and IL-33, yet the molecular mechanisms and cooperative interaction between these cytokines remain unclear. Our objective was to investigate the molecular mechanism and cooperation of IL-4 and IL-33 in eosinophil activation. Eosinophils derived from bone marrow or isolated from Il5-transgenic mice were activated in the presence of IL-4 or IL-33 for 1 or 4 h, and the transcriptome was analyzed by RNA sequencing. The candidate genes were validated by quantitative PCR and ELISA. We demonstrated that murine-cultured eosinophils respond to IL-4 and IL-33 by phosphorylation of STAT-6 and NF-κB, respectively. RNA sequence analysis of murine-cultured eosinophils indicated that IL-33 induced 519 genes, whereas IL-4 induced only 28 genes, including 19 IL-33-regulated genes. Interestingly, IL-33 induced eosinophil activation via two distinct mechanisms, IL-4 independent and IL-4 secretion/autostimulation dependent. Anti-IL-4 or anti-IL-4Rα Ab-treated cultured and mature eosinophils, as well as Il4- or Stat6-deficient cultured eosinophils, had attenuated protein secretion of a subset of IL-33-induced genes, including Retnla and Ccl17. Additionally, IL-33 induced the rapid release of preformed IL-4 protein from eosinophils by a NF-κB-dependent mechanism. However, the induction of most IL-33-regulated transcripts (e.g., Il6 and Il13) was IL-4 independent and blocked by NF-κB inhibition. In conclusion, we have identified a novel activation pathway in murine eosinophils that is induced by IL-33 and differentially dependent upon an IL-4 auto-amplification loop.
authors.affiliation: Division of Allergy and Immunology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229.
authors.forename: Carine
authors.initials: C
authors.lastname: Bouffi
authors.forename: Mark
authors.initials: M
authors.lastname: Rochman
authors.forename: Christopher B
authors.initials: CB
authors.lastname: Zust
authors.forename: Emily M
authors.initials: EM
authors.lastname: Stucke
authors.forename: Andrey
authors.initials: A
authors.lastname: Kartashov
authors.forename: Patricia C
authors.initials: PC
authors.lastname: Fulkerson
authors.forename: Artem
authors.initials: A
authors.lastname: Barski
authors.forename: Marc E
authors.initials: ME
authors.lastname: Rothenberg
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology; D001483:Base Sequence; D001854:Bone Marrow Cells / Q000276:immunology*; D002478:Cells, Cultured; D004804:Eosinophils / Q000276:immunology*; D007249:Inflammation / Q000276:immunology; D000067596:Interleukin-33; D015847:Interleukin-4 / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D007378:Interleukins / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism; D010766:Phosphorylation; D019948:Receptors, Interleukin-4 / Q000276:immunology; D050800:STAT6 Transcription Factor / Q000235:genetics / Q000378:metabolism; D017423:Sequence Analysis, RNA; D015398:Signal Transduction / Q000276:immunology
chemical_list: D000906:Antibodies; C506535:Il33 protein, mouse; D000067596:Interleukin-33; D007378:Interleukins; D016328:NF-kappa B; D019948:Receptors, Interleukin-4; D050800:STAT6 Transcription Factor; C494100:Stat6 protein, mouse; D015847:Interleukin-4
references.citation: J Immunol. 2006 Jun 15;176(12):7456-61
references.pmid: 16751391
references.citation: Am J Pathol. 2011 Aug;179(2):733-44
references.pmid: 21699881
references.citation: Int Immunol. 2010 Jun;22(6):479-89
references.pmid: 20501612
references.citation: J Immunol. 2009 Nov 15;183(10):6469-77
references.pmid: 19841166
references.citation: Nat Protoc. 2012 Mar 01;7(3):562-78
references.pmid: 22383036
references.citation: Proc Natl Acad Sci U S A. 2008 May 20;105(20):7240-5
references.pmid: 18480254
references.citation: J Immunol. 2008 Feb 15;180(4):2443-9
references.pmid: 18250453
references.citation: Nature. 2002 Dec 19-26;420(6917):825-9
references.pmid: 12490951
references.citation: Traffic. 2005 Nov;6(11):1047-57
references.pmid: 16190985
references.citation: J Immunol. 2005 Jun 1;174(11):7111-22
references.pmid: 15908341
references.citation: J Immunol. 2010 Sep 15;185(6):3472-80
references.pmid: 20693421
references.citation: Bioinformatics. 2004 Jun 12;20(9):1453-4
references.pmid: 14871861
references.citation: J Immunol. 2007 Aug 15;179(4):2051-4
references.pmid: 17675461
references.citation: J Exp Med. 1986 May 1;163(5):1085-99
references.pmid: 3486243
references.citation: Immunity. 1996 Mar;4(3):313-9
references.pmid: 8624821
references.citation: Allergy. 2012 Sep;67(9):1118-26
references.pmid: 22702477
references.citation: Proc Natl Acad Sci U S A. 2013 Jun 11;110(24):9914-9
references.pmid: 23716700
references.citation: J Immunol. 1998 Sep 15;161(6):3128-35
references.pmid: 9743380
references.citation: J Immunol. 1992 Feb 15;148(4):1086-92
references.pmid: 1371130
references.citation: J Immunol. 2008 Sep 15;181(6):4004-9
references.pmid: 18768855
references.citation: Science. 1998 Dec 18;282(5397):2258-61
references.pmid: 9856949
references.citation: Lab Invest. 2008 Nov;88(11):1245-53
references.pmid: 18762778
references.citation: Nat Rev Immunol. 2010 Feb;10(2):103-10
references.pmid: 20081870
references.citation: Circ Res. 2010 Sep 3;107(5):650-8
references.pmid: 20634488
references.citation: J Allergy Clin Immunol. 2010 Mar;125(3):752-4
references.pmid: 20153038
references.citation: Immunity. 2004 Mar;20(3):267-77
references.pmid: 15030771
references.citation: Annu Rev Immunol. 2006;24:147-74
references.pmid: 16551246
references.citation: Am J Pathol. 2004 Apr;164(4):1315-26
references.pmid: 15039219
references.citation: PLoS One. 2012;7(6):e39673
references.pmid: 22761864
references.citation: J Immunol. 2012 Mar 15;188(6):2622-9
references.pmid: 22327070
references.citation: J Immunol. 2008 Nov 1;181(9):5981-9
references.pmid: 18941187
references.citation: Bioinformatics. 2004 Nov 22;20(17):3246-8
references.pmid: 15180930
references.citation: J Allergy Clin Immunol. 1998 Nov;102(5):783-8
references.pmid: 9819295
references.citation: J Exp Med. 1991 Sep 1;174(3):745-8
references.pmid: 1875171
references.citation: J Immunol. 2009 Oct 15;183(8):5094-103
references.pmid: 19801525
references.citation: J Exp Med. 2006 Jun 12;203(6):1435-46
references.pmid: 16702603
references.citation: Proc Natl Acad Sci U S A. 2006 Feb 28;103(9):3333-8
references.pmid: 16492782
references.citation: Proc Natl Acad Sci U S A. 1999 Aug 17;96(17):9660-5
references.pmid: 10449750
references.citation: J Asthma. 2008 Oct;45(8):648-53
references.pmid: 18951255
references.citation: Cell Signal. 2008 Sep;20(9):1679-86
references.pmid: 18603409
references.citation: Cell. 2013 Apr 11;153(2):376-88
references.pmid: 23582327
references.citation: Blood. 2009 Feb 12;113(7):1526-34
references.pmid: 18955562
references.citation: Cell Mol Immunol. 2010 Jan;7(1):26-34
references.pmid: 20029461
references.citation: J Immunol. 2009 Oct 1;183(7):4403-14
references.pmid: 19734227
references.citation: J Allergy Clin Immunol. 2006 Aug;118(2):410-9
references.pmid: 16890766
references.citation: Science. 2011 Apr 8;332(6026):243-7
references.pmid: 21436399
references.citation: J Allergy Clin Immunol. 2008 Jun;121(6):1484-90
references.pmid: 18539196
references.citation: J Leukoc Biol. 2008 Apr;83(4):817-21
references.pmid: 18192490
references.citation: J Allergy Clin Immunol. 2008 Dec;122(6):1200-1207.e1
references.pmid: 19084112
references.citation: J Immunol. 2009 Feb 15;182(4):2357-63
references.pmid: 19201890
references.citation: Trends Mol Med. 2004 Oct;10(10):493-9
references.pmid: 15464449
references.citation: J Allergy Clin Immunol. 2001 Feb;107(2):295-301
references.pmid: 11174196
references.citation: Immunity. 2005 Nov;23(5):479-90
references.pmid: 16286016
references.citation: Immunology. 2013 Sep;140(1):70-7
references.pmid: 23582173
references.citation: J Immunol. 2012 Feb 1;188(3):1075-82
references.pmid: 22190185