Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cells
<p>Abstract</p> <p>Background</p> <p>Cytotoxic T lymphocytes (CTL) and natural killer (NK) cells play an essential role in the host defence against intracellular pathogens such as <it>Listeria</it>, and <it>Mycobacteria</it>. The key mediator of...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2007-08-01
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| Series: | BMC Immunology |
| Online Access: | http://www.biomedcentral.com/1471-2172/8/14 |
| _version_ | 1819054803701989376 |
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| author | Wagner Carsten A Dumrese Claudia Sundstrom Hanna Velic Ana Latinovic-Golic Sonja Walch Michael Groscurth Peter Ziegler Urs |
| author_facet | Wagner Carsten A Dumrese Claudia Sundstrom Hanna Velic Ana Latinovic-Golic Sonja Walch Michael Groscurth Peter Ziegler Urs |
| author_sort | Wagner Carsten A |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>Cytotoxic T lymphocytes (CTL) and natural killer (NK) cells play an essential role in the host defence against intracellular pathogens such as <it>Listeria</it>, and <it>Mycobacteria</it>. The key mediator of bacteria-directed cytotoxicity is granulysin, a 9 kDa protein stored in cytolytic granules together with perforin and granzymes. Granulysin binds to cell membranes and is subsequently taken up via a lipid raft-associated mechanism. In dendritic cells (DC) granulysin is further transferred via early endosomes to <it>L. innocua</it>-containing phagosomes were bacteriolysis is induced. In the present study we analysed the role of perforin in granulysin-induced intracellular bacteriolysis in DC.</p> <p>Results</p> <p>We found granulysin-induced lysis of intracellular <it>Listeria </it>significantly increased when perforin was simultaneously present. In pulse-chase experiments enhanced bacteriolysis was observed when perforin was added up to 25 minutes after loading the cells with granulysin demonstrating no ultimate need for simultaneous uptake of granulysin and perforin. The perforin concentration sufficient to enhance granulysin-induced intracellular bacteriolysis did not cause permanent membrane pores in <it>Listeria</it>-challenged DC as shown by dye exclusion test and LDH release. This was in contrast to non challenged DC that were more susceptible to perforin lysis. For Listeria-challenged DC, there was clear evidence for an Ca<sup>2+ </sup>influx in response to sublytic perforin demonstrating a short-lived change in the plasma membrane permeability. Perforin treatment did not affect granulysin binding, initial uptake or intracellular trafficking to early endosomes. However, enhanced colocalization of granulysin with listerial DNA in presence of perforin was found by confocal laser scanning microscopy.</p> <p>Conclusion</p> <p>The results provide evidence that perforin increases granulysin-mediated killing of intracellular <it>Listeria </it>by enhanced phagosome-endosome fusion triggered by a transient Ca<sup>2+ </sup>flux.</p> |
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| format | Article |
| id | doaj.art-265f78c963264e2cb1c29e8c134d1e94 |
| institution | Directory Open Access Journal |
| issn | 1471-2172 |
| language | English |
| last_indexed | 2024-12-21T12:57:26Z |
| publishDate | 2007-08-01 |
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| series | BMC Immunology |
| spelling | doaj.art-265f78c963264e2cb1c29e8c134d1e942022-12-21T19:03:17ZengBMCBMC Immunology1471-21722007-08-01811410.1186/1471-2172-8-14Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cellsWagner Carsten ADumrese ClaudiaSundstrom HannaVelic AnaLatinovic-Golic SonjaWalch MichaelGroscurth PeterZiegler Urs<p>Abstract</p> <p>Background</p> <p>Cytotoxic T lymphocytes (CTL) and natural killer (NK) cells play an essential role in the host defence against intracellular pathogens such as <it>Listeria</it>, and <it>Mycobacteria</it>. The key mediator of bacteria-directed cytotoxicity is granulysin, a 9 kDa protein stored in cytolytic granules together with perforin and granzymes. Granulysin binds to cell membranes and is subsequently taken up via a lipid raft-associated mechanism. In dendritic cells (DC) granulysin is further transferred via early endosomes to <it>L. innocua</it>-containing phagosomes were bacteriolysis is induced. In the present study we analysed the role of perforin in granulysin-induced intracellular bacteriolysis in DC.</p> <p>Results</p> <p>We found granulysin-induced lysis of intracellular <it>Listeria </it>significantly increased when perforin was simultaneously present. In pulse-chase experiments enhanced bacteriolysis was observed when perforin was added up to 25 minutes after loading the cells with granulysin demonstrating no ultimate need for simultaneous uptake of granulysin and perforin. The perforin concentration sufficient to enhance granulysin-induced intracellular bacteriolysis did not cause permanent membrane pores in <it>Listeria</it>-challenged DC as shown by dye exclusion test and LDH release. This was in contrast to non challenged DC that were more susceptible to perforin lysis. For Listeria-challenged DC, there was clear evidence for an Ca<sup>2+ </sup>influx in response to sublytic perforin demonstrating a short-lived change in the plasma membrane permeability. Perforin treatment did not affect granulysin binding, initial uptake or intracellular trafficking to early endosomes. However, enhanced colocalization of granulysin with listerial DNA in presence of perforin was found by confocal laser scanning microscopy.</p> <p>Conclusion</p> <p>The results provide evidence that perforin increases granulysin-mediated killing of intracellular <it>Listeria </it>by enhanced phagosome-endosome fusion triggered by a transient Ca<sup>2+ </sup>flux.</p>http://www.biomedcentral.com/1471-2172/8/14 |
| spellingShingle | Wagner Carsten A Dumrese Claudia Sundstrom Hanna Velic Ana Latinovic-Golic Sonja Walch Michael Groscurth Peter Ziegler Urs Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cells BMC Immunology |
| title | Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cells |
| title_full | Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cells |
| title_fullStr | Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cells |
| title_full_unstemmed | Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cells |
| title_short | Perforin enhances the granulysin-induced lysis of Listeria innocua in human dendritic cells |
| title_sort | perforin enhances the granulysin induced lysis of listeria innocua in human dendritic cells |
| url | http://www.biomedcentral.com/1471-2172/8/14 |
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