An in-vitro model of intracellular bacterial infection using the murine macrophage cell line J774.2

A simple model of intracellular bacterial infection based on the ability of the macrophage cell line J774.2 to phagocytose Escherichia coli in a reproducible manner is described. Viable counting of intracellular bacteria and microscopic examination of infected macrophage cultures after treatment with fluorescent E. coli antibody showed that the bacteria multiplied within the J774.2 cells. Viable intracellular bacteria may be used to study the activity of bactericidal and bacteriostatic drugs within the macrophage. The low apparent intracellular bactericidal activity of streptomycin, which was time- and concentration-dependent, accords with a low permeability of the J774.2 macrophages to this antibiotic. An exclusively intracellular infection could be achieved by inactivation of non-phagocytosed extracellular bacteria by streptomycin treatment of infected macrophage cultures. Under appropriate conditions intracellular bacterial viability was unaffected.