A crucial role for TNF-alpha in mediating neutrophil influx induced by endogenously generated or exogenous chemokines, KC/CXCL1 and LIX/CXCL5.
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.
Vieira
S M
SM
Department of Pharmacology, School of Medicine of Ribeirão Preto, University of São Paulo, São Paulo, Brazil.
Lemos
H P
HP
Grespan
R
R
Napimoga
M H
MH
Dal-Secco
D
D
Freitas
A
A
Cunha
T M
TM
Verri
W A
WA
Souza-Junior
D A
DA
Jamur
M C
MC
Fernandes
K S
KS
Oliver
C
C
Silva
J S
JS
Teixeira
M M
MM
Cunha
F Q
FQ
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*
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
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