Contribution of cytokine-induced neutrophil chemoattractant CINC-2 and CINC-3 to neutrophil recruitment in lipopolysaccharide-induced inflammation in rats.
OBJECTIVE
We have estimated the contribution of three types of cytokine-induced neutrophil chemoattractants (CINC-1, -2 and -3) and rat macrophage inflammatory protein (MIP)-1alpha to neutrophil recruitment in the air pouch/ lipopolysaccharide (LPS)-induced inflammation in rats.


METHODS
Excess amounts of anti-CINC-2, CINC-3 and MIP-1alpha antibodies (Abs), together with LPS (1 microg/ml), were injected into the preformed air pouch of rats. Chemokine levels and chemotactic activity in the pouch fluid were measured using enzyme-linked immunosorbent assay and multiwell-type Boyden chambers in vitro, respectively.


RESULTS
Excess amounts of the Abs significantly neutralized CINC-1 and almost completely neutralized CINC-2, CINC-3 and MIP-1 alpha in the pouch fluid, and a significant suppression (about 60% inhibition) of neutrophil infiltration by the Abs was found. In agreement with the in vivo results, in vitro neutralization experiments demonstrated that complete neutralization of CINCs and MIP-1alpha by the Abs resulted in a marked suppression (73% inhibition) of chemotactic potency of 8-h pouch fluid (exudate) from LPS-treated rats. On the other hand, treatment with anti-CINC-2, CINC-3 or MIP-1alpha Ab alone resulted in 48%, 34% or 10% inhibition, respectively, of chemotactic activity of the 8-h exudate.


CONCLUSIONS
Our results suggest that CINC-2, a novel rat CXC chemokine and CINC-3 play an important role in neutrophil recruitment in the rat air pouch/LPS-induced inflammation.
Takano
K
K
Department of Physiological Chemistry, Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, Japan. takanok@ms.toyama-mpu.ac.jp
Nakagawa
H
H
D000388:Air; D000818:Animals; D019138:Antibodies, Blocking / Q000494:pharmacology; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; D002630:Chemotactic Factors / Q000037:antagonists & inhibitors / Q000276:immunology / Q000502:physiology*; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D006133:Growth Substances / Q000276:immunology / Q000502:physiology*; D007074:Immunoglobulin G / Q000276:immunology; D007249:Inflammation / Q000139:chemically induced / Q000473:pathology*; D036341:Intercellular Signaling Peptides and Proteins; D008070:Lipopolysaccharides; D019402:Macrophage Inflammatory Proteins / Q000502:physiology; D008297:Male; D009504:Neutrophils / Q000502:physiology*; D051381:Rats; D017208:Rats, Wistar
D019138:Antibodies, Blocking; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D054360:Chemokine CXCL1; D019743:Chemokines, CXC; D002630:Chemotactic Factors; C513297:Cxcl1 protein, rat; C513294:Cxcl3 protein, rat; D006133:Growth Substances; D007074:Immunoglobulin G; D036341:Intercellular Signaling Peptides and Proteins; D008070:Lipopolysaccharides; D019402:Macrophage Inflammatory Proteins