PROTEIN-TYROSINE KINASE BUT NOT PROTEIN-KINASE-C INHIBITION BLOCKS RECEPTOR-INDUCED ALVEOLAR MACROPHAGE ACTIVATION

THE selective enzyme inhibitors genistein and Ro 31-8220 were used to assess the importance of protein tyrosine kinase (PTR) and protein kinase C (PKC), respectively, in N-formyl-methionyl-leucyl-phenylalanine (FMLP) induced generation of superoxide anion and thromboxane B-2 (TXB(2)) in guinea-pig alveolar macrophages (AM). Genistein (3-100 mu M) dose dependently inhibited FMLP (3 nM) induced superoxide generation in non-primed AM and TXB(2) release in non-primed or in lipopolysaccharide (LPS) (10 ng/ml) primed AM to a level > 80% but had little effect up to 100 mu M on phorbol myristate acetate (PMA) (10 nM) induced superoxide release. Ro 31-8220 inhibited PMA induced superoxide generation (IC50 0.21 +/- 0.10 mu M) but had no effect on or potentiated (at 3 and 10 mu M) FMLP responses in non-primed AM. In contrast, when present during LPS priming as well as during FMLP challenge Ro 31-8220 (10 mu M) inhibited primed TXB(2) release by > 80%. The results indicate that PTK activation is required for the generation of these inflammatory mediators by FMLP in AM. PRC activation appears to be required for LPS priming but not for transducing the FMLP signal; rather, PKC activation may modulate the signal by a negative feedback mechanism.