title: Activation of mitogen-activated protein kinases during granulocyte apoptosis in patients with severe sepsis.
abstract: Reduction of neutrophil apoptosis represents a major cause for granulocytosis and increases the destructive potential of theses cells during systemic inflammatory response syndrome (SIRS) and sepsis. In this light, the role of protein kinases for the regulation of altered neutrophil apoptosis under infectious conditions was investigated. Neutrophils, obtained from patients with severe sepsis (n = 18), were incubated ex vivowith either LPS (1 microg/mL) or interferon-gamma (IFN-gamma; 10 ng/mL) for 16 h. Apoptosis was determined by propidium iodine (PI) staining of DNA fragments and was compared with the rate of spontaneous apoptosis. Tyrosine kinases were inhibited by herbimycin (1 microM), the mitogen-activated protein (MAP) kinase ERK was inhibited with PD98059 (50 microM), and p38 MAP kinase was inhibited with SB203580 (5 microM). Herbimycin reconstituted LPS-reduced apoptosis in neutrophils from controls (39.9 +/- 3.8%) and patients (20.8 +/- 2.8%) to levels seen in spontaneous apoptosis (70.9 +/- 2.8% and 40.7 +/- 3.7%, respectively). Inhibition of the ERK kinase yielded similar results, whereas SB203580 had no effect on LPS-reduced apoptosis. However, inhibition of p38 partially reconstituted IFN-gamma-reduced apoptosis (51.3 +/- 7.7% and 25.6 +/- 5.8%) and increased spontaneous apoptosis (82.4 +/- 3.3% and 42.0 +/- 5.8%) in controls and patients, respectively. Western blot analysis revealed phosphorylation of both MAP kinases by LPS, but not by IFN-gamma. Inhibition of MAP kinases did not augment neutrophil apoptosis in patients to the level seen in controls, indicating that other mechanisms must be involved in the regulation of neutrophil apoptosis. Although the ERK kinase regulates LPS-induced reduction of apoptosis, the p38 MAP kinase might be involved in IFN-gamma signaling and the feedback regulation of neutrophil apoptosis.
authors.affiliation: Division of Trauma Surgery, University Hospital Zürich, Switzerland.
authors.forename: Luc
authors.initials: L
authors.lastname: Härter
authors.forename: Marius
authors.initials: M
authors.lastname: Keel
authors.forename: Ursula
authors.initials: U
authors.lastname: Steckholzer
authors.forename: Udo
authors.initials: U
authors.lastname: Ungethuem
authors.forename: Otmar
authors.initials: O
authors.lastname: Trentz
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Ertel
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D017209:Apoptosis / Q000187:drug effects; D016227:Benzoquinones; D016022:Case-Control Studies; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D006098:Granulocytes / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology*; D006801:Humans; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000494:pharmacology; D047029:Lactams, Macrocyclic; D008070:Lipopolysaccharides / Q000494:pharmacology; D020935:MAP Kinase Signaling System; D008297:Male; D008875:Middle Aged; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010766:Phosphorylation; D011809:Quinones / Q000494:pharmacology; D011994:Recombinant Proteins; D017828:Rifabutin / Q000031:analogs & derivatives; D018805:Sepsis / Q000201:enzymology* / Q000473:pathology*; D018746:Systemic Inflammatory Response Syndrome / Q000201:enzymology / Q000473:pathology; D048051:p38 Mitogen-Activated Protein Kinases
chemical_list: D016227:Benzoquinones; D004791:Enzyme Inhibitors; D047029:Lactams, Macrocyclic; D008070:Lipopolysaccharides; D011809:Quinones; D011994:Recombinant Proteins; D017828:Rifabutin; C020754:herbimycin; D007371:Interferon-gamma; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases