The Journal of Experimental Medicine
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© The Rockefeller University Press, 0022-1007/1997/10/1159/ $5.00
The Journal of Experimental Medicine, Volume 186, Number 7, October 6, 1997 1159-1163

Brief Definitive Reports

pH-dependent Perforation of Macrophage Phagosomes by Listeriolysin O from Listeria monocytogenes

Kathryn E. Beauregard*,{ddagger}, Kyung-Dall Lee*, R. John Collier{ddagger}, and Joel A. Swanson*

From the * Department of Cell Biology and {ddagger} Department of Microbiology and Molecular Genetics, Harvard Medical School, Boston, Massachusetts 02115

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.

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: jswan{at}umich.edu. K.-D. Lee's current address is the College of Pharmacy, University of Michigan, Ann Arbor, MI 48109.


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