Appearance of cytokine-induced neutrophil chemoattractant isoforms and immunolocalization of them in lipopolysaccharide-induced acute lung inflammation in rats

Abstract.

Objective and Design: Recently, rat cytokine-induced neutrophil chemoattractant (CINC), which belongs to the interleukin-8 family, was grouped into four isoforms, CINC-1, CINC-2α, CINC-2β, and CINC-3. To determine the major component and the source of CINC in airways, we investigated the change in appearance of CINC isoforms after exposure of rats to lipopolysaccharide. ¶Methods: Male Sprague-Dawley rats, 8-10 weeks old, were used in the present study. Bronchoalveolar lavage (BAL) was performed at 1, 2, 4, 6, 12, and 24 h after lipopolysaccharide inhalation (4 mg/ml for 30 min). The concentrations of each specific rat CINC in the BAL supernatant were measured by use of commercially available kits. Furthermore, lung tissue was employed for immunohistochemical staining of CINCs (CINC-1, -2α, -2β, and -3) using the streptavidin-biotin technique. ¶Results: Inhalation of lipopolysaccharide caused increases in CINC-1, CINC-2α, and CINC-3 in BAL fluids, whereas CINC-2β was not detected. The increases in CINC-2α and CINC-3 were less than the increase in CINC-1. Positive immunohistochemical staining for CINC-1 was detected in bronchial noncilliated cells and in certain neutrophils that had infiltrated into the submucosa. ¶Conclusions: These findings suggest that CINC-1 is the major isoform among the four CINCs in lipopolysaccharide-induced acute lung inflammation in rats. Its sources are likely to be bronchial noncilliated cells and certain infiltrating neutrophils.