title: Bacterial phagocytosis activates extracellular signal-regulated kinase and p38 mitogen-activated protein kinase cascades in human neutrophils.
abstract: The hypothesis that bacterial phagocytosis by human polymorphonuclear neutrophils (PMNs) stimulates MAPK cascades that regulate respiratory burst activation was tested. Extracellular response kinase (ERK) and p38 kinase, but not c-Jun NH2-terminal kinase, activities were increased within 5 min of phagocytosis of plasma-opsonized Staphylococcus aureus (S-SA), reached maximum at 20-30 min, and remained elevated through 60 min. The role of Fcy receptors was examined using gamma globulin-opsonized SA (IgG-SA), whereas CR3 receptors were activated by particulate beta-glucan. IgG-SA stimulated a maximal ERK activity at 30 min, whereas p38 activity was maximal at 5 min. Beta-glucan stimulated maximal ERK activity at 5 min and maximal p38 activity at 2 min. Non-opsonized bacteria were ingested at 10% of the level of S-SA and stimulated a minimal increase in ERK and p38 activity at 60 min. S-SA stimulation of ERK was inhibited by wortmannin, LY294002, and genistein, but not calphostin C; whereas p38 stimulation was inhibited by calphostin C and genistein, but not wortmannin and LY294002. Simultaneous measurement of phagocytosis and H2O2 production by flow cytometry was used to assess the role of ERKs and p38 kinase in phagocytosis. The MEK inhibitor PD098059 had no significant effect on phagocytosis or H2O2 production. The p38 kinase inhibitor SB203580 significantly attenuated H2O2 production, whereas phagocytosis was unaffected. In conclusion, bacterial phagocytosis stimulates ERK and p38 activation by distinct signal transduction pathways. Phagocytosis-stimulated p38 kinase activity is necessary for optimal H2O2 production.
authors.affiliation: Department of Medicine, University of Louisville Health Sciences Center, Veterans Affairs Medical Center, Kentucky, USA. kmcleish@e-mail.kdp-baptist.louisville.edu
authors.forename: K R
authors.initials: KR
authors.lastname: McLeish
authors.forename: J B
authors.initials: JB
authors.lastname: Klein
authors.forename: P Y
authors.initials: PY
authors.lastname: Coxon
authors.forename: K Z
authors.initials: KZ
authors.lastname: Head
authors.forename: R A
authors.initials: RA
authors.lastname: Ward
mesh_terms: D015703:Antigens, CD / Q000502:physiology; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism* / Q000502:physiology; D004789:Enzyme Activation; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases; D016177:Macrophage-1 Antigen / Q000502:physiology; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism / Q000502:physiology; D009504:Neutrophils / Q000201:enzymology* / Q000382:microbiology* / Q000502:physiology; D010587:Phagocytosis / Q000502:physiology*; D017452:Receptors, IgG / Q000502:physiology; D016897:Respiratory Burst / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology*; D013211:Staphylococcus aureus / Q000502:physiology*; D048051:p38 Mitogen-Activated Protein Kinases
chemical_list: D015703:Antigens, CD; C080568:Fc gamma receptor IIA; D016177:Macrophage-1 Antigen; D017452:Receptors, IgG; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D048031:JNK Mitogen-Activated Protein Kinases; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases