Possible roles of protein kinases in neutrophil chemotactic factor production by leucocytes in allergic inflammation in rats

Abstract

1 In an air pouch-type allergic inflammation model in rats, leucocytes that had infiltrated into the pouch fluid collected 4 h after the antigen challenge produced proteinaceous chemotactic factors for neutrophils when they were incubated in the medium. 2 To clarify the mechanism of activation of the infiltrated leucocytes in producing these factors, the effects of protein kinase inhibitors on neutrophil chemotactic factor production were examined. 3 When the infiltrated leucocytes were incubated for 4 h in medium containing the non-selective protein kinase inhibitor K-252a (1-100 ng ml-', 2.14-214 nM), the tyrosine kinase inhibitor genistein (1-50 1tgml', 3.7-185 p M), and the more selective protein kinase C inhibitor H-7 (5-100 A gml', 13.7-274 gM); neutrophil chemotactic activity in the conditioned medium was decreased in a concentration-dependent manner, but the adenosine 3':5'-cyclic monophosphate (cAMP)-dependent protein kinase inhibitor H-89 (1-10Ongml1', 2.24-2240nM) showed no effect.4 Isoelectric focusing of the conditioned medium revealed that the leucocytes produced two neutrophil chemotactic factors, leucocyte-derived neutrophil chemotactic factor (LDNCF) 1 and LDNCF-2. Treatment of the leucocytes with K-252a, genistein, and H-7, but not H-89, inhibited production of both LDNCF-1 and LDNCF-2. 5 These results suggest that activation of tyrosine kinase and protein kinase C, but not cAMPdependent protein kinase, is responsible for the production of LDNCF-1 and LDNCF-2. 6 The steroidal anti-inflammatory drug dexamethasone and the protein synthesis inhibitor cycloheximide inhibited neutrophil chemotactic factor production in a concentration-dependent manner. Timecourse experiments showed that the inhibitory effect by dexamethasone was apparent even 30 min after the incubation. 7 Mechanism for inhibiting the production of LDNCF-1 and LDNCF-2 by dexamethasone is also discussed.

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

“…These results indicate that the leukocytes from the immunized rats pro duce higher amounts of MIP-2 than the leukocytes from the nonimmunized rats. Recently we found that the leukocytes infiltrated into the pouch fluid of the immunized rats pro duce LDNCF-1 and LDNCF-2 much more actively than do the leukocytes from the nonimmunized rats through the mechanism of activation of tyrosine kinase and protein ki nase C but not cAMP-dependent protein kinase [16], Be cause LDNCF-2 has been identified as rat MIP-2 in this study, the mechanism of action of the protein kinase inhib itors on LDNCF-2 production by the leukocytes might be able to analyze in the gene expression level.…”

Neuronal and Glial Apoptosis after Traumatic Spinal Cord Injury

“…However, the possibility cannot be presently excluded that the beneficial effects of cycloheximide observed here were mediated by mechanisms not related to direct inhibition of programmed cell death. Some alternative mechanisms might be enhancement of cellular glutathione levels because of reduced cysteine use (Ratan et al, 1994) or suppression of neutrophil chemotaxis (Tanabe et al, 1994). Yet another possible mechanism is raised by the interesting finding that low concentrations of cycloheximide that produce only limited inhibition of protein synthesis can induce the production of Bcl2 and antioxidant enzymes (Furukawa et al, 1997).…”

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.…”

“…Rat cytokine-induced neutrophil chemoattractants (CINCs) are the gro-gene products consisting of CINC-1, CINC-2a, CINC-2b, and CINC-3 [Nakagawa et al, 1994]. 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].…”

“…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].…”