We previously reported that treatment of mice with a neutralizing mAb against listeriolysin O (LLO), the pore-forming toxin of Listeria monocytogenes, provided resistance to this intracellular bacterium. We evaluated whether anti-LLO mAb would affect Listeria handling by macrophages, essential cells in Listeria resistance. Macrophages infected in the presence of anti-LLO mAb showed a marked reduction in intracellular Listeria growth, with a concomitant block in LLO-dependent Listeria passage from phagosome to cytosol. Anti-LLO mAb did not opsonize Listeria but, rather, acted within macrophages to neutralize LLO. Importantly, anti-LLO mAb effects on Listeria growth were independent of Fc gamma receptor expression, IFN gamma signaling, and production of nitric oxide and superoxide. These results identify a novel mechanism for antibody control of bacteria within macrophages.