Intratracheal administration of endotoxin and cytokines. VI. Antiserum to CINC inhibits acute inflammation

Abstract

Cytokine-induced neutrophil chemoattractant (CINC), a chemotactic molecule of the interleukin (IL)-8 family, is known to be induced in the rat in response to tumor necrosis factor (TNF), IL-1, and lipopolysaccharide (LPS). Intratracheal injection of endotoxin (LPS) is shown to cause CINC mRNA expression in pulmonary tissue, peaking after 2 h, and CINC protein expression in bronchoalveolar lavage (BAL) fluid, peaking after 2-4 h. Intratracheal injection of synthetic CINC causes acute inflammation that is abrogated by coinjection of antiserum to purified natural rat CINC. Intratracheal injection of antiserum to CINC inhibits intratracheal LPS- and IL-1-induced neutrophil emigration into BAL fluid by approximately 60-70%. Despite the anti-inflammatory activity of anti-CINC antiserum, TNF is elevated in the lavage fluid of rats receiving anti-CINC, suggesting that CINC may act in a negative feedback loop to downregulate TNF expression. Intratracheal injection of antiserum to CINC combined with intravenous injection of anti-E-selectin antibody inhibits intratracheal LPS- and IL-1-induced neutrophil emigration into BAL fluid by approximately 75-85%. CINC-mediated chemotactic activity and E-selectin-mediated adherence of neutrophils to endothelium contribute significantly to the pathogenesis of LPS-initiated acute inflammation.

The Effects of Granulocyte Colony-Stimulating Factor and Neutrophil Recruitment on the Pulmonary Chemokine Response to Intratracheal Endotoxin

“…AMs were cultured for 6 h in the presence of LPS (50 ng/ml). Pulmonary recruited PMNs were isolated from G-CSF-treated rats challenged with intratracheal LPS for 4 h. CINC, two members from the C-X-C chemokine family, have been shown to be responsible for the major chemotactic activity for PMNs in the alveolar space in rats (15,16). In this study, we determined the effects of G-CSF pretreatment on the expression of these chemokines in the lung following intratracheal challenge with LPS.…”

“…MIP-2 and CINC are potent C-X-C chemokines accounting for the major chemotactic activity of the rat lung for PMNs (15,16). We determined the pulmonary chemokine responses following an intratracheal challenge with LPS in rats pretreated with or without G-CSF.…”

Differential Effects of Ebselen on Neutrophil Recruitment, Chemokine, and Inflammatory Mediator Expression in a Rat Model of Lipopolysaccharide-Induced Pulmonary Inflammation

“…Reverse transcription. A 1-g portion of total RNA was subjected to first-strand cDNA synthesis in a 25-l reaction mixture containing avian myeloblastosis virus reverse transcriptase (10 U), dNTP mixture (2 mM concentrations of each dNTP), oligo(dT) [12][13][14][15][16][17][18] primers (10 M), and reaction buffer as supplied with the enzyme (50 mM Tris-HCl (pH 8.3), 50 mM KCl, 10 mM MgCl 2 , 0.5 mM spermidine, and 10 mM DDT). The samples were incubated in a PerkinElmer 480 thermal cycler (PerkinElmer, Wellesley, MA) at 42°C for 60 min followed by enzyme denaturation step at 94°C for 2 min.…”

“…TNF-␣ has been implicated as a mediator of LPS-induced airway inflammation (12,13,15,19). TNF-␣ was shown to initiate and amplify pulmonary inflammatory responses by stimulating the release of chemotactic factors by up-regulating the expression of leukocyte and endothelial adhesion molecules (16,23).…”

“…The mechanism of pulmonary PMN migration in rats given intratracheal LPS has been partially characterized and involves production by airway cells of inflammatory cytokines such as TNF-␣ and IL-1␤ as well as neutrophil-specific chemoattractants known as CXC chemokines, which in rats include the family of cytokineinduced neutrophil chemoattractants (CINCs) and macrophage-inflammatory protein-2 (MIP-2) also known as CINC-3 (12)(13)(14)(15)(16)(17)(18)(19). CD18 integrins have also been shown to play a role for full polymorphonuclear cell (PMN) migration.…”