The mechanism of neutrophil chemotactic factor production and the characteristics of the neutrophil chemotactic factor produced by the infiltrated leukocytes in allergic inflammation.
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Leukocytes collected from the pouch fluid 4 h after the antigen challenge in the air pouch-type allergic inflammation model in rats produced proteinaceous chemotactic factors for neutrophils when they were incubated. The chemotactic factor production by the leukocytes from the immunized rats was significantly higher than that from the non-immunized rats. The production of the neutrophil chemotactic factors was inhibited by the nonspecific protein kinase inhibitor K-252a, the protein kinase C inhibitor H-7, and the tyrosine kinase inhibitor genistein, but was not inhibited by the cAMP-dependent protein kinase inhibitor H-89, suggesting that protein kinase C and tyrosine kinase but not cAMP-dependent protein kinase are activated in the leukocytes infiltrated into the inflammatory site to produce the neutrophil chemotactic factors.<BR>Isoelectric focusing of the neutrophil chemotactic factors in the conditioned medium revealed that the leukocytes produce two factors, leukocyte derived neutrophil chemotactic factor (LDNCF) -1 and LDNCF-2, of which the pI values are 4-5 and >8, respectively. LDNCF-2, a major component of the neutrophil chemotactic factors, was purified by gel chromatography and reverse-phase HPLC, and the Nterminal amino acid sequence of the purified LDNCF-2 was determined.<BR>It was demonstrated that the sequence (1 to 20) was identical to the 32 to 51 amino acid sequence of the proform of the rat macrophage inflammatory protein (MIP) -2. Furthermore, RT-PCR revealed that the level of MIP-2 mRNA was higher in the leukocytes from the immunized rats than that from the non-immunized rats.
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