Effect of growth condition on in-vitro susceptibility of Shigella dysenteriae type 1 killing by murine peritoneal macrophages

Abstract

The intracellular fate of Shigella dysenteriae type 1 strains grown in casamino acid-yeast extract (CYE) broth and nutrient broth (NB) was studied in casein-elicited mouse peritoneal macrophages. Virulent strains 14731 and W30864 cultured in NB and opsonised with normal mouse serum were susceptible to killing by peritoneal macrophages (66 SEM 1.7% killing by 2 h). In contrast, both strains grown in CYE broth and opsonised with normal mouse serum showed resistance to killing by peritoneal macrophages (76 SEM 1.4% survival by 2 h). Electronmicroscopy demonstrated that the bacteria escaped from the phagosome compartment by lysing the phagocytic vacuole and remained within the cytoplasm. Lipopolysaccharide (LPS) stimulated peritoneal macrophages to kill the opsonised strains 14731 and W30864 grown in CYE broth (85.4 SEM 1.6% killing by 2 h). Recombinant murine gamma interferon (rIFN-gamma) also stimulated macrophages to kill CYE-grown bacteria (52.1 SEM 1.3% killing by 2 h). However, an avirulent rough mutant strain W30864-22 grown in either NB or CYE broth showed marked susceptibility to killing by peritoneal macrophages, which was similar to that of NB-grown strain 14731 or W30864. The results of the present study suggest that in-vitro growth conditions may modulate the susceptibility of S. dysenteriae type 1 to killing by phagocytes