Effect of purified staphylococcal leukocidal toxins on isolated blood polymorphonuclear leukocytes and peritoneal macrophages in vitro

Abstract

Bicomponent (fractions S and F) staphylococcal leukocidal toxins (Panton-Valentine leukocidin-Luk and haemolysin gamma-Hlg) were tested for in vitro activity against isolated polymorphonuclear leukocytes (PMNL) and peritoneal macrophages (PMF). For assessment of membrane permeability at subcytolytic concentrations of leukocidin (Luk-S + Luk-F) and haemolysin gamma (HlgA + HlgB) (8-1000 ng/ml), PMNL and PMF were radiolabelled (86Rb, 14C-amino-isobutyric acid (AIB) or 51Cr). All toxins tested caused lysis of human PMNL, although considerable differences were noted in the sensitivity of these cells to Luk and Hlg. Release of 51Cr (at 1000-5000 ng/ml), being a sign of irreversible cell damage and lysis, was preceded, at lower concentrations of the toxins (40 and 200 ng/ml), by the release of large amounts of low-molecular labels--86Rb and 14C-AIB. In another experiment, it was found that release of 86Rb from PMNL incubated with low concentrations of Luk (50 ng/ml) took place after 15-30 minutes of incubation, when no significant amounts of 14C-AIB or 51Cr were released. These findings support the concept of pore formation by staphylococcal leukocidal toxins in membranes of sensitive cells and indicate that a relatively short time is needed for the formation of these pores after binding of the Luk-S and Luk-F components to the membrane.