ABSTRACT Pneumolysin is a thiol-activated membrane-damaging toxin produced by Streptococcus pneumoniae . The toxin plays a role in virulence of the pneumococcus in animal models of infection. Pneumolysin has a range of biological activity including the ability to lyse eukaryotic cells and to interfere with the function of cells and soluble molecules of the immune system. The use of purified native and mutant toxin and of isogenic mutants of the pneumococcus expressing altered versions of the toxin has allowed the contribution of the various activities of this multifunctional toxin to virulence to be defined.