Role of superoxide dismutase and catalase as determinants of pathogenicity of Nocardia asteroides: importance in resistance to microbicidal activities of human polymorphonuclear neutrophils

Abstract

The roles of nocardial superoxide dismutase (SOD) and catalase in the resistance of Nocardia asteroides to the microbicidal properties of human polymorphonuclear leukocytes were determined in vitro. The neutrophils killed ca. 80% of the cells of the less virulent N. asteroides 10905 and ca. 50% of the log phase of the more virulent N. asteroides GUH-2 after 180 min of incubation. These phagocytes were not able to kill early-stationary-phase cells of strain GUH-2 that contained 10 times more intracytoplasmic catalase than log-phase cells of the same culture. However, the polymorphonuclear leukocytes were able to kill more than 50% of the cells of early-stationary-phase strain GUH-2 after treatment with purified antibody specific for surface-associated SOD. No killing was observed when the bacteria were treated with normal rabbit immunoglobulin G or with serum obtained from rabbits immunized against whole nocardial cells (containing little or no activity against SOD). These phagocytes killed more than 99% of Listeria monocytogenes used as a control. Chlorpromazine-treated polymorphonuclear leukocytes killed L. monocytogenes (70%) but they were not able to kill antibody-treated cells of N. asteroides GUH-2. Exogenously added SOD partially protected strain 10905, which lacked surface-associated enzyme, but it had no effect on the killing of strain GUH-2, which already possessed significant amounts of surface-bound SOD. In contrast, catalase added to the nocardiae provided almost complete protection to the log-phase cells of strain GUH-2, but strain 10905 was only partially protected. SOD combined with catalase had additive activity which completely protected the cells of strain 10905. A mutant of N. asteroides GUH-2 (SCII-C) is more virulent during the log phase than is the parental strain. This mutant contained at least 7 times more catalase at this stage of growth than did the parent. No other differences between these two strains were observed during the log phase. In sharp contrast to those of the parent, log-phase cells of this high-catalase mutant were not killed by polymorphonuclear phagocytes. These data indicate a role for both SOD and catalase in the resistance of Nocardia spp. to human neutrophils, and they represent at least two factors associated with virulence.