The pore-forming toxin listeriolysin O (LLO) is a major virulence factor implicated in escape of Listeria monocytogenes from phagocytic vacuoles. Here we describe the pH-dependence of vacuolar perforation by LLO, using the membrane-impermeant fluorophore 8-hydroxypyrene-1,3,6-trisulfonic acid (HPTS) to monitor the pH and integrity of vacuoles in mouse bone marrow–derived macrophages. Perforation was observed when acidic vacuoles containing wild-type L. monocytogenes displayed sudden increases in pH and release of HPTS into the cytosol. These changes were not seen with LLO-deficient mutants. Perforation occurred at acidic vacuolar pH (4.9–6.7) and was reduced in frequency or prevented completely when macrophages were treated with the lysosomotropic agents ammonium chloride or bafilomycin A1. We conclude that acidic pH facilitates LLO activity in vivo.
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6 October 1997
Brief Definitive Report|
October 06 1997
pH-dependent Perforation of Macrophage Phagosomes by Listeriolysin O from Listeria monocytogenes
Kathryn E. Beauregard,
Kathryn E. Beauregard
From the *Department of Cell Biology and ‡Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115
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Kyung-Dall Lee,
Kyung-Dall Lee
From the *Department of Cell Biology and ‡Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115
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R. John Collier,
R. John Collier
From the *Department of Cell Biology and ‡Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115
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Joel A. Swanson
Joel A. Swanson
From the *Department of Cell Biology and ‡Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115
Search for other works by this author on:
Kathryn E. Beauregard
From the *Department of Cell Biology and ‡Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115
Kyung-Dall Lee
From the *Department of Cell Biology and ‡Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115
R. John Collier
From the *Department of Cell Biology and ‡Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115
Joel A. Swanson
From the *Department of Cell Biology and ‡Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115
Address correspondence to Dr. Joel A. Swanson, Department of Anatomy and Cell Biology, University of Michigan Medical School, 1335 Catherine Ave., Ann Arbor, MI 48109-0616. Phone: 313-647-6339; FAX: 313-763-1166; E-mail: [email protected]. K.-D. Lee's current address is the College of Pharmacy, University of Michigan, Ann Arbor, MI 48109.
Received:
April 09 1997
Revision Received:
August 08 1997
Online ISSN: 1540-9538
Print ISSN: 0022-1007
1997
J Exp Med (1997) 186 (7): 1159–1163.
Article history
Received:
April 09 1997
Revision Received:
August 08 1997
Citation
Kathryn E. Beauregard, Kyung-Dall Lee, R. John Collier, Joel A. Swanson; pH-dependent Perforation of Macrophage Phagosomes by Listeriolysin O from Listeria monocytogenes . J Exp Med 6 October 1997; 186 (7): 1159–1163. doi: https://doi.org/10.1084/jem.186.7.1159
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