| Induction of apoptosis in macrophage cell line, J774, by the cell-free supernatant from Pseudomonas aeruginosa. | |
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MedLine Citation:
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PMID: 12725289 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Pseudomonas aeruginosa is able to secrete many virulence factors that are cytotoxic towards eukaryotic cells. To investigate the effect of the bacterium on macrophages, we obtained cell-free supernatants from P. aeruginosa (Pa) IID1117 (elastase-positive and protease-positive) and Pa IID1130 (elastase-positive and protease-negative). After 6 hr of incubation with the cell-free supernatant from the Pa IID1117 strain, the viability of J774 macrophages was shown to be significantly reduced (47.5+/-11%), but not Pa IID1130 (96.4+/-1.6%) at a concentration of 10% (v/v) compared to control J774 macrophages without any supernatant (97.2+/-1.7%) by the detection of trypan blue dye exclusion. The death of cells was further demonstrated to be due to apoptosis characterized by chromatin condensation and apoptotic bodies by Hoechst 33258 staining, DNA fragmentation by agarose gel electrophoresis and terminal deoxynucleotidyl transferase-mediated d-UTP nick end labeling (TUNEL). An activated subunit was found to be released from procaspase-3 in cell lysate. But in the presence of protease inhibitor, the apoptosis was completely blocked. The findings indicate that the Pa IID1117 strain is capable of inducing apoptosis in J774 macrophages. The apoptosis induced by the cell-free supernatant from Pa IID1117 strain is suggested to be dependent on protease, but not elastase. |
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Authors:
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Jianling Zhang; Hisao Takayama; Takashi Matsuba; Ru Jiang; Yoshinori Tanaka |
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Publication Detail:
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Type: Journal Article |
Journal Detail:
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Title: Microbiology and immunology Volume: 47 ISSN: 0385-5600 ISO Abbreviation: Microbiol. Immunol. Publication Date: 2003 |
Date Detail:
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Created Date: 2003-05-02 Completed Date: 2003-07-11 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 7703966 Medline TA: Microbiol Immunol Country: Japan |
Other Details:
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Languages: eng Pagination: 199-206 Citation Subset: IM |
Affiliation:
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Department of Microbiology and Pathology, School of Life Science, Faculty of Medicine, Tottori University, Yonago, Tottori 683-8503, Japan. |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Apoptosis* Caspase 3 Caspases / metabolism Cell Line Cells, Cultured Culture Media Macrophages / cytology, drug effects*, metabolism Protease Inhibitors / pharmacology Pseudomonas aeruginosa / metabolism, pathogenicity* |
| Chemical | |
Reg. No./Substance:
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0/Culture Media; 0/Protease Inhibitors; EC 3.4.22.-/Caspase 3; EC 3.4.22.-/Caspases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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