COMPLEMENTATION OF A DNAK-DEFICIENT ESCHERICHIA-COLI STRAIN WITH THE DNAK/DNAJ OPERON OF BRUCELLA-OVIS REDUCES THE RATE OF INITIAL INTRACELLULAR KILLING WITHIN THE MONOCYTIC CELL-LINE U937

Facultatively intracellular bacteria express heat shock proteins after phagocytosis by macrophages. Using non-pathogenic Escherichia coli strains and the human monocytic cell line U937, we showed that deletion of the dnaK gene significantly increased the rate of initial intracellular killing of bacteria. Trans-complementation of the deletion mutant with the dnaK/dnaJ operon of Brucella ovis restored the pattern of intracellular elimination of the control strain expressing dnaK. These differences were not observed using antibody-opsonized bacteria and activated cells. In vitro, strains expressing dnaK resisted hydrogen peroxide better than the deletion mutant; in contrast, the mutant complemented by dnaK/dnaJ of B. ovis tolerated low pH and low H2O2 better than the wild-type strain and the deletion mutant. Our results suggested the participation of DnaK in protection of intracellular bacteria against antimicrobial macrophage factors.