Characterization of Leukocyte-Derived Neutrophil Chemotactic Factor-2 and Its Possible Roles in Neutrophil Infiltration in Allergic Inflammation in Rats

Abstract

This study sought to clarify the responsible chemotactic factor for neutrophils in allergic inflammation in rats. When the leukocytes collected from the pouch fluid 4 h after injection of the antigen solution into the air pouch were incubated, the neutrophil chemotactic activity in the conditioned medium increased time-dependently with higher levels for the leukocytes from the immunized rats than the nonimmunized ones. The chemotactic activity did not result from cytokine-induced neutrophil chemoattractant (CINC) because CINC concentrations in the conditioned medium were low. Neutrophil chemotactic factors in the conditioned medium were separated by isoelectric focusing into two factors, leukocyte-derived neutrophil chemotactic factor (LDNCF)-1 and LDNCF-2. The activity of LDNCF-2 was more than 75% of the total chemotactic activity in the conditioned medium. LDNCF-2 was purified by gel chromatography and reverse-phase HPLC. The N-terminal amino acid sequence (1–20) of the purified LDNCF-2 was identical to the 32-51 amino acid sequence of the pro-form of rat macrophage inflammatory protein (MIP)-2. Higher levels of MIP-2 mRNA in the leukocytes from the immunized rats than that from the non-immunized rats were proved by the reverse transcription-polymerase chain reaction. In vivo, concentrations of CINC in the pouch fluid were low, and did not represent the chemotactic activity in the pouch fluid. These results suggest that LDNCF-2 (MIP-2) is an important chemotactic factor for neutrophils in the allergic inflammation in rats.

Differential Production of Chemokines and Their Role in Neutrophil Infiltration in Rat: Allergic Inflammation

“…The results do not rule out the possibility that another CINC is the major chemokine in a different type of rat inflammation model, because we demonstrated that CINC-2α is the major chemoattractant produced by granulation tissue [5] or lipopolysaccharide-stimulated rat macrophages [6] in culture. In contrast to our results, Tanabe et al [16] demonstrated that CINC-1 was a minor chemokine, and CINC-3/rMIP-2 was the major one in the pouch fluid (exudate) of azobenzenearsonate-conjugated acetyl bovine serum albumin (ABA-AcBSA)-induced allergic inflammation in rats. The differences between the ABA-AcBSA- and FITC-OVA-induced allergic inflammation models may be due to the intensity of inflammation and cell types surrounding air pouch (inflammatory site).…”

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

“…Nakagawa demonstrated that CINC2a was the major chemokine produced by granulation tissus [8] or LPS-stimulated rat macrophages in vitro [25]. In an allergic inflammation model utilizing azobenzenearsonateconjugated acetyl bovine serum albumin, Tanabe et al found that CINC-3 was the major chemokine and CINC-1 was a minor one [30]; whereas Nakagawa reported CINC-1 to be the major chemokine and CINC-3 to be a minor one in a fluorescein isothiocyanate-labeled ovalbumin-induced inflammation model [31]. These reports suggest that the major CINC isoform derived from an inflammatory response might depend on the stimulator.…”

Analysis of the mechanism regulating the stability of rat macrophage inflammatory protein‐2 mRNA in RBL‐2H3 cells

“…4). Therefore, it is possible that the IgE-sensitized mast cells in the air pouch-type allergic inflammation model participate in the production of MIP-2 and contribute to neutrophil infiltration in this model [Tanabe et al, 1994[Tanabe et al, , 1995aXiao et al, 1997]. Furthermore, we suggested that p38 MAPK and p44/42 MAPK play important roles in mediating the antigen-induced MIP-2 mRNA stabilization, because the p38 MAPK inhibitor SB203580 and the MEK inhibitor PD98059 lowered the antigen-induced retardation of MIP-2 mRNA decay (Fig.…”

“…CINC-3 is also known as macrophage inflammatory protein-2 (MIP-2) and leucocyte-derived neutrophil chemotactic factor-2 (LDNCF-2) [Tanabe et al, 1994[Tanabe et al, , 1995c. In the rat air pouch-type allergic inflammation model, MIP-2 (CINC-3, LDNCF-2) plays an important role in neutrophil infiltration [Tanabe et al, 1994[Tanabe et al, , 1995aXiao et al, 1997]. Previously, we reported that staurosporine, generally used as a non-specific protein kinase inhibitor [Tamaoki et al, 1986], stimulates MIP-2 production in rat peritoneal neutrophils [Xiao et al, 1999].…”