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| Item Type: | Article |
|---|---|
| Title: | Phosphatidylinositol 3-kinase controls antineutrophil cytoplasmic antibodies - Induced respiratory burst in human neutrophils |
| Creators Name: | Kettritz, R., Choi, M., Butt, W., Rane, M., Rolle, S., Luft, F.C. and Klein, J.B. |
| Abstract: | Antineutrophil cytoplasmic antibodies (ANCA) activate human polymorphonuclear neutrophils (PMN) primed with tumor necrosis factor alpha (TNF-{alpha}) in vitro. Phosphatidylinositol 3-kinase (PI3-K) and the protein-serine/threonine kinase Akt have been implicated in the control of the phagocyte respiratory burst. The hypothesis that PI3-K controls the ANCA-induced respiratory burst was tested. TNF-{alpha}-primed PMN were stimulated with a monoclonal antibody to myeloperoxidase (MPO) and with PR3- and MPO-ANCA, respectively. Akt activation was assessed with phospho-specific antibodies. Superoxide release was measured with ferricytochrome. ANCA antigen translocation was assessed by fluorescence-activated cell sorter. The effect of TNF-{alpha} and MPO-ANCA on Akt signaling was studied with immunoprecipitation and glutathione S-transferase pull-down assays. Western blotting revealed rapid transient Akt phosphorylation during TNF-{alpha} priming and a second phosphorylation after ANCA. PI3-K inhibition by LY294002 blocked both Akt phosphorylation and superoxide generation. A total of 20 +/- 3 nmol O(2)(-)/0.75 x 10(6) PMN/45 min was released after stimulation with PR3-ANCA. LY294002 (5 microM) decreased this amount to 0.3 +/- 2.6 nmol (n = 10, P < 0.05); the MPO-ANCA values were 23 +/- 3 versus 1.6 +/- 3.6 (n = 10, P < 0.05). p38 MAPK inhibition with 10 microM SB202190 that also decreased ANCA-induced superoxide generation prevented S473 phosphorylation of Akt in response to TNF-{alpha} and to ANCA. However, SB202190 but not LY294002 abrogated TNF-{alpha}-mediated ANCA antigen surface translocation, demonstrating that superoxide generation and ANCA antigen translocation proceed by separate mechanisms. Akt, PAK1, and Rac1 existed as cytosolic complex in resting PMN. TNF-alpha stimulation increased association of PAK1 with Akt. An MPO monoclonal antibody did not alter the Akt signaling complex further. The data demonstrate the importance of PI3-K for the ANCA-induced PMN oxidant production. |
| Keywords: | Antigens, Antineutrophil Cytoplasmic Antibodies, Biological Transport, Chromones, Enzyme Inhibitors, Mitogen-Activated Protein Kinases, Morpholines, Neutrophils, Phosphatidylinositol 3-Kinases, Protein-Serine-Threonine Kinases, Proto-Oncogene Proteins, Proto-Oncogene Proteins c-akt, Respiratory Burst, Signal Transduction, Tumor Necrosis Factor-alpha, p38 Mitogen-Activated Protein Kinases |
| Source: | Journal of the American Society of Nephrology |
| ISSN: | 1046-6673 |
| Publisher: | American Society of Nephrology |
| Volume: | 13 |
| Number: | 7 |
| Page Range: | 1740-1749 |
| Date: | 1 July 2002 |
| Official Publication: | https://doi.org/10.1097/01.ASN.0000019411.36000.06 |
| PubMed: | View item in PubMed |
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