Open AccessAccumulation of staphylococcal P anton– V alentine leukocidin in the detergent‐resistant membrane microdomains on the target cells is essential for its cytotoxicity
Author(s) -
Nishiyama Akihito,
Isobe Hirokazu,
Iwao Yasuhisa,
Takano Tomomi,
Hung WeiChun,
Taneike Ikue,
Nakagawa Saori,
Dohmae Soshi,
Iwakura Nobuhiro,
Yamamoto Tatsuo
Publication year - 2012
Publication title -
fems immunology & medical microbiology
Language(s) - English
Resource type - Journals
eISSN - 1574-695X
pISSN - 0928-8244
DOI - 10.1111/j.1574-695x.2012.01027.x
Subject(s) - cytotoxicity , leukocidin , microbiology and biotechnology , panton–valentine leukocidin , biology , lysis , incubation , immune system , staphylococcus aureus , immunology , biochemistry , bacteria , in vitro , methicillin resistant staphylococcus aureus , genetics
The mechanisms for the cytotoxicity of staphylococcal P anton– V alentine leukocidin ( PVL ), a pore‐forming toxin consisting of L uk S ‐ PV and L uk F ‐ PV , in human immune cells are still unclear. Because L uk S ‐ PV binds to ganglioside GM 1, a constituent of detergent‐resistant membrane microdomains ( DRM s) of the plasma membrane, the role of DRM s in PVL cytotoxicity was examined in human polymorphonuclear neutrophils ( PMN s), monocytes, HL ‐60 cells, and THP ‐1 cells. PVL binding capacities in HL ‐60 and THP ‐1 cells were higher than those in PMN s and monocytes; however, the PVL concentration to obtain more than 80% cell lysis in HL ‐60 cells was 10 times higher than that in PMN s and PVL even at such concentration induced < 10% cell lysis in THP ‐1 cells. After incubation of PMN s with L uk S ‐ PV , more than 90% of L uk S ‐ PV bound to the detergent‐soluble membranes. Subsequent incubation with L uk F ‐ PV at 4 °C induced the accumulation of more than 70% of PVL components and 170‐ to 220‐kDa complex formation in DRM s in an actin‐independent manner. However, only 30% of PVL was found, and complex formation was under detectable level in DRM s in HL ‐60 cells. PVL did not accumulate in DRM s in THP ‐1 cells. Our observations strongly indicate that PVL accumulation in DRM s is essential for PVL cytotoxicity.