Role of intracellular kinases in the regulation of equine eosinophil migration and actin polymerization.
Inappropriately activated eosinophils can contribute to disease pathogenesis and intracellular signalling pathways that regulate functional responses may represent a therapeutic target. Little is known about intracellular signalling in equine eosinophils and this study examined the role of phospholipase C (PLC) and a range of protein kinases on responses to histamine and CCL11. Histamine (10(-4) M) or CCL11 (5.6 x 10(-9) M)-induced actin polymerization, migration and superoxide production by eosinophils from healthy horses were compared in the presence and absence of selective kinase inhibitors. Inhibition of phosphatidylinositol-3 kinase (PI3K) significantly reduced the response in each assay. In contrast, whilst inhibition of PLC decreased actin polymerization and superoxide production, an increase in migration was observed; the latter effect was also seen when protein kinase C (PKC) was inhibited. With the exception of histamine-induced migration, which was significantly reduced by blocking extracellular regulated kinase (ERK)1/2, activation of ERK1/2, p38 MAPK and tyrosine kinase did not appear to play an important role in the responses studied. These results suggest that equine eosinophil activation by histamine and CCL11 is mediated through PI3K. Whilst PLC activation is required for actin polymerization and superoxide production, migration may be negatively regulated by PLC and PKC. These kinases represent potential targets for modulating eosinophil activation by multiple stimuli.
Weston
M C
MC
Department of Veterinary Basic Sciences and Pathology and Infectious Diseases, The Royal Veterinary College, Hertfordshire, UK. marie.weston@piramed.com
Collins
M E
ME
Cunningham
F M
FM
D000199:Actins / Q000187:drug effects*; D000818:Animals; D002465:Cell Movement / Q000187:drug effects; D002478:Cells, Cultured; D054413:Chemokine CCL11 / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D004804:Eosinophils / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D004963:Estrenes / Q000494:pharmacology; D006632:Histamine / Q000494:pharmacology; D006736:Horses / Q000502:physiology*; D007211:Indoles / Q000494:pharmacology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D011760:Pyrrolidinones / Q000494:pharmacology; D015398:Signal Transduction / Q000502:physiology; D010738:Type C Phospholipases / Q000037:antagonists & inhibitors* / Q000494:pharmacology
D000199:Actins; D054413:Chemokine CCL11; D004791:Enzyme Inhibitors; D004963:Estrenes; D007211:Indoles; D011760:Pyrrolidinones; C060229:1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione; D006632:Histamine; D011493:Protein Kinase C; D010738:Type C Phospholipases; C064758:Ro 31-8220