title: A crucial role for TNF-alpha in mediating neutrophil influx induced by endogenously generated or exogenous chemokines, KC/CXCL1 and LIX/CXCL5.
abstract: OBJECTIVE
  Chemokines orchestrate neutrophil recruitment to inflammatory foci. In the present study, we evaluated the participation of three chemokines, KC/CXCL1, MIP-2/CXCL2 and LIX/CXCL5, which are ligands for chemokine receptor 2 (CXCR2), in mediating neutrophil recruitment in immune inflammation induced by antigen in immunized mice.
  
  
  METHODS
  Neutrophil recruitment was assessed in immunized mice challenged with methylated bovine serum albumin, KC/CXCL1, LIX/CXCL5 or tumour necrosis factor (TNF)-alpha. Cytokine and chemokine levels were determined in peritoneal exudates and in supernatants of macrophages and mast cells by elisa. CXCR2 and intercellular adhesion molecule 1 (ICAM-1) expression was determined using immunohistochemistry and confocal microscopy.
  
  
  RESULTS
  Antigen challenge induced dose- and time-dependent neutrophil recruitment and production of KC/CXCL1, LIX/CXCL5 and TNF-alpha, but not MIP-2/CXCL2, in peritoneal exudates. Neutrophil recruitment was inhibited by treatment with reparixin (CXCR1/2 antagonist), anti-KC/CXCL1, anti-LIX/CXCL5 or anti-TNF-alpha antibodies and in tumour necrosis factor receptor 1-deficient mice. Intraperitoneal injection of KC/CXCL1 and LIX/CXCL5 induced dose- and time-dependent neutrophil recruitment and TNF-alpha production, which were inhibited by reparixin or anti-TNF-alpha treatment. Macrophages and mast cells expressed CXCR2 receptors. Increased macrophage numbers enhanced, while cromolyn sodium (mast cell stabilizer) diminished, LIX/CXCL5-induced neutrophil recruitment. Macrophages and mast cells from immunized mice produced TNF-alpha upon LIX/CXCL5 stimulation. Methylated bovine serum albumin induced expression of ICAM-1 on mesenteric vascular endothelium, which was inhibited by anti-TNF-alpha or anti-LIX/CXCL5.
  
  
  CONCLUSIONS
  Following antigen challenge, CXCR2 ligands are produced and act on macrophages and mast cells triggering the production of TNF-alpha, which synergistically contribute to neutrophil recruitment through induction of the expression of ICAM-1.
authors.affiliation: Department of Pharmacology, School of Medicine of Ribeirão Preto, University of São Paulo, São Paulo, Brazil.
authors.forename: S M
authors.initials: SM
authors.lastname: Vieira
authors.forename: H P
authors.initials: HP
authors.lastname: Lemos
authors.forename: R
authors.initials: R
authors.lastname: Grespan
authors.forename: M H
authors.initials: MH
authors.lastname: Napimoga
authors.forename: D
authors.initials: D
authors.lastname: Dal-Secco
authors.forename: A
authors.initials: A
authors.lastname: Freitas
authors.forename: T M
authors.initials: TM
authors.lastname: Cunha
authors.forename: W A
authors.initials: WA
authors.lastname: Verri
authors.forename: D A
authors.initials: DA
authors.lastname: Souza-Junior
authors.forename: M C
authors.initials: MC
authors.lastname: Jamur
authors.forename: K S
authors.initials: KS
authors.lastname: Fernandes
authors.forename: C
authors.initials: C
authors.lastname: Oliver
authors.forename: J S
authors.initials: JS
authors.lastname: Silva
authors.forename: M M
authors.initials: MM
authors.lastname: Teixeira
authors.forename: F Q
authors.initials: FQ
authors.lastname: Cunha
mesh_terms: D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D002417:Cattle; D054360:Chemokine CXCL1 / Q000276:immunology* / Q000494:pharmacology; D054365:Chemokine CXCL5 / Q000276:immunology* / Q000494:pharmacology; D018799:Intercellular Adhesion Molecule-1 / Q000096:biosynthesis; D017737:Macrophages, Peritoneal / Q000276:immunology / Q000378:metabolism; D008407:Mast Cells / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009504:Neutrophils / Q000276:immunology*; D010538:Peritonitis / Q000276:immunology* / Q000378:metabolism; D023062:Receptors, Interleukin-8A / Q000037:antagonists & inhibitors; D023063:Receptors, Interleukin-8B / Q000037:antagonists & inhibitors / Q000096:biosynthesis; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000235:genetics; D012709:Serum Albumin / Q000276:immunology; D013449:Sulfonamides / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000276:immunology / Q000502:physiology*
chemical_list: D000906:Antibodies; D054360:Chemokine CXCL1; D054365:Chemokine CXCL5; C475895:Cxcl1 protein, mouse; C094739:Cxcl5 protein, mouse; D023062:Receptors, Interleukin-8A; D023063:Receptors, Interleukin-8B; D047888:Receptors, Tumor Necrosis Factor, Type I; D012709:Serum Albumin; D013449:Sulfonamides; C506236:Tnfrsf1a protein, mouse; D014409:Tumor Necrosis Factor-alpha; D018799:Intercellular Adhesion Molecule-1; C490707:reparixin
references.citation: Immunology. 1991 Jan;72(1):65-72
references.pmid: 1997402
references.citation: J Clin Invest. 2006 Mar;116(3):695-702
references.pmid: 16485040
references.citation: J Immunol. 1997 Oct 1;159(7):3595-602
references.pmid: 9317159
references.citation: J Immunol. 2007 Dec 1;179(11):7891-8
references.pmid: 18025236
references.citation: Eur Respir J. 2008 Apr;31(4):773-82
references.pmid: 18094004
references.citation: J Biol Chem. 2007 Apr 20;282(16):11658-66
references.pmid: 17197447
references.citation: Immunology. 1991 Aug;73(4):472-7
references.pmid: 1916898
references.citation: J Allergy Clin Immunol. 2009 Feb;123(2):486-92
references.pmid: 18986691
references.citation: J Clin Invest. 1996 Apr 15;97(8):1952-9
references.pmid: 8621780
references.citation: Cytokine. 2008 Feb;41(2):174-81
references.pmid: 18180165
references.citation: J Leukoc Biol. 2005 Sep;78(3):716-24
references.pmid: 15961577
references.citation: Inflamm Res. 2007 Jun;56(6):230-9
references.pmid: 17607547
references.citation: Proc Am Thorac Soc. 2006 May;3(3):245-51
references.pmid: 16636093
references.citation: Arthritis Rheum. 2008 Aug;58(8):2329-37
references.pmid: 18668539
references.citation: Neoplasia. 2008 Mar;10(3):244-54
references.pmid: 18320069
references.citation: J Histochem Cytochem. 1997 Dec;45(12):1715-22
references.pmid: 9389775
references.citation: Regul Pept. 2007 Jan 10;138(1):40-8
references.pmid: 17014919
references.citation: Eur J Immunol. 2004 Nov;34(11):2955-63
references.pmid: 15376193
references.citation: Int Arch Allergy Immunol. 1997 Jan;112(1):27-35
references.pmid: 8980461
references.citation: BMC Cancer. 2008 Jun 25;8:178
references.pmid: 18578857
references.citation: J Exp Med. 2000 Nov 20;192(10):1441-52
references.pmid: 11085746
references.citation: J Leukoc Biol. 1999 Feb;65(2):249-55
references.pmid: 10088608
references.citation: Cytokine Growth Factor Rev. 1998 Jun;9(2):167-73
references.pmid: 9754710
references.citation: Proc Natl Acad Sci U S A. 2005 Feb 1;102(5):1755-60
references.pmid: 15665080
references.citation: J Immunol. 2002 Dec 1;169(11):6467-73
references.pmid: 12444156
references.citation: Int J Immunopharmacol. 1994 Sep;16(9):703-10
references.pmid: 7806429
references.citation: Br J Pharmacol. 2004 Sep;143(1):132-42
references.pmid: 15302676
references.citation: Cytokine. 2005 Jul 7;31(1):9-17
references.pmid: 15967374
references.citation: Inflammation. 2008 Feb;31(1):36-46
references.pmid: 17874178
references.citation: Immunobiology. 2008;213(7):533-44
references.pmid: 18656701
references.citation: J Histochem Cytochem. 2001 Feb;49(2):219-28
references.pmid: 11156690
references.citation: Mol Immunol. 2006 Mar;43(7):897-914
references.pmid: 16084593
references.citation: Inflammation. 1984 Jun;8 Suppl:S3-14
references.pmid: 6090313
references.citation: N Engl J Med. 1998 Feb 12;338(7):436-45
references.pmid: 9459648
references.citation: J Immunol Methods. 1991 Jan 24;136(1):11-6
references.pmid: 1995703
references.citation: Eur J Immunol. 2002 Feb;32(2):393-403
references.pmid: 11813158
references.citation: Arthritis Rheum. 2001 Jan;44(1):31-40
references.pmid: 11212173
references.citation: Immunogenetics. 2002 Nov;54(8):599-603
references.pmid: 12439624
references.citation: Immunology. 2008 Dec;125(4):591-600
references.pmid: 18710405
references.citation: Blood. 2005 Jun 1;105(11):4282-9
references.pmid: 15718418
references.citation: Blood. 1997 Aug 1;90(3):909-28
references.pmid: 9242519
references.citation: J Biol Chem. 1989 Aug 5;264(22):13267-72
references.pmid: 2526814
references.citation: Int Arch Allergy Immunol. 2002 Jun;128(2):142-50
references.pmid: 12065915
references.citation: Arthritis Rheum. 2008 Jul;58(7):1968-73
references.pmid: 18576313
references.citation: Eur J Immunol. 2008 May;38(5):1215-24
references.pmid: 18412166
references.citation: Br J Pharmacol. 2001 Dec;134(8):1619-28
references.pmid: 11739237
references.citation: J Exp Med. 1977 Sep 1;146(3):857-68
references.pmid: 894190
references.citation: Immunology. 2008 Jul;124(3):295-303
references.pmid: 18217950
references.citation: Curr Drug Targets Inflamm Allergy. 2005 Jun;4(3):273-9
references.pmid: 16101533
references.citation: Arthritis Rheum. 2008 Jul;58(7):2030-40
references.pmid: 18576322
references.citation: J Biol Chem. 2003 Feb 14;278(7):4675-86
references.pmid: 12468547
references.citation: J Diabetes Complications. 2009 May-Jun;23(3):178-84
references.pmid: 18413205
references.citation: Pancreatology. 2008;8(4-5):488-97
references.pmid: 18765953
references.citation: J Clin Invest. 1989 Oct;84(4):1045-9
references.pmid: 2677047
references.citation: Am J Physiol Heart Circ Physiol. 2004 May;286(5):H1750-6
references.pmid: 14704234