Salmonella enterica serovar Typhimurium periplasmic superoxide dismutases SodCI and SodCII are required for protection against the phagocyte oxidative burst - PubMed (original) (raw)

Salmonella enterica serovar Typhimurium periplasmic superoxide dismutases SodCI and SodCII are required for protection against the phagocyte oxidative burst

Laura M Sly et al. Infect Immun. 2002 Sep.

Abstract

Vitamin D(3) (1,25-dihydroxycholecalciferol) induced the phagocyte oxidative burst and intracellular killing of Salmonella enterica serovar Typhimurium in a phosphatidylinositol 3-kinase-dependent manner. The antimicrobial effect was more pronounced for Salmonella SodCI and SodCII mutants, confirming the role of the phagocyte oxidase in the vitamin D(3) effect. The results for an in vitro system with human THP-1 cells correlate with in vivo virulence data for mice and show that both the SodCI and SodCII enzymes are required to protect against the oxidative burst.

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Figures

FIG. 1.

FIG. 1.

Vitamin D3 restricts the intracellular growth of wild-type S. enterica serovar Typhimurium and induces the phagocyte oxidative burst in a PI 3-kinase-dependent manner. (A) Intracellular S. enterica serovar Typhimurium quantitated by serial dilution in Luria-Bertani broth and plating on Luria-Bertani agar 0, 2, 4, and 8 h after infection of THP-1 cells. Cells were either not treated or treated with the PI 3-K inhibitor Wm (50 nM) or Ly (15 μM) and then were left untreated or treated with vitamin D3 (100 nM). The data are averages from three independent experiments performed in triplicate. An asterisk indicates that the P value was <0.03 when the growth in vitamin D3-treated cells was compared to the growth in control cells or when the growth in cells treated with Ly or Wm plus vitamin D3 was compared to the growth in cells treated only with vitamin D3, as determined by analysis of variance at each time point. (B) Superoxide secreted by macrophages in 1 h determined by measuring the Sod-inhibitable reduction of ferricytochrome c in the presence and absence of Salmonella infection, vitamin D3 treatment, and the inhibitor Ly or Wm. An asterisk indicates that the P value was <0.02 when the data for Salmonella infection and vitamin D3 treatment was compared to the data for either treatment alone or when the data for pretreatment with Ly or Wm followed by Salmonella infection and vitamin D3 treatment was compared to the data for no pretreatment, as determined by an unpaired Student's t test.

FIG. 2.

FIG. 2.

S. enterica serovar Typhimurium Sod mutants are more susceptible to the antimicrobial activity of vitamin D3 than the wild-type strain. Differentiated THP-1 cells were infected with wild-type S. enterica serovar Typhimuriu_m_ (wt) or with a SodCI, SodCII, or double SodCI-SodCII S. enterica serovar Typhimurium mutant and then were left untreated or were treated with vitamin D3. The CFU of intracellular bacteria were enumerated 0, 4, and 8 h after infection. The data are averages from three independent experiments performed in triplicate. An asterisk indicates that the P value was <0.05 when the growth of each mutant in vitamin D3-treated cells was compared to the growth in control cells, as determined by analysis of variance at each time point.

FIG. 3.

FIG. 3.

Killing of intracellular S. enterica serovar Typhimurium Sod mutants by vitamin D3 is PI 3-K dependent. Differentiated THP-1 cells were infected with the SodCI mutant (A), the SodCII mutant (B), and the SodCI-SodCII double mutant (C). Cells either were left untreated or were treated with Wm or Ly and then were either left untreated or treated with vitamin D3. The CFU of intracellular bacteria were enumerated 0, 4, and 8 h after infection. The data are from three independent experiments performed in triplicate. An asterisk indicates that the P value was <0.04 (A), <0.05 (B), or <0.01 (C) when growth in vitamin D3-treated cells was compared to growth in control cells or when the data for pretreatment with Ly or Wm followed by Salmonella infection and vitamin D3 treatment of cells was compared to the data for no pretreatment of cells. Comparisons were made by performing analysis of variance at each time point.

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