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Regulation of mammalian siderophore 2,5-DHBA in the innate immune response to infection.

Liu Z, Reba S, Chen WD, Porwal SK, Boom WH, Petersen RB, Rojas R, Viswanathan R, Devireddy L - J. Exp. Med. (2014)

Bottom Line: Pathogens secrete small iron-binding moieties, siderophores, to acquire host iron.In support of this idea, supplementation with mammalian siderophore enhances bacterial growth in vitro.Thus, reciprocal regulation of 24p3 and mammalian siderophore is a protective mechanism limiting microbial access to iron.

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Affiliation: Case Comprehensive Cancer Center; Department of Pathology; Department of Medicine, Tuberculosis Research Institute and Division of Infectious Diseases; and Department of Chemistry, Case Western Reserve University, Cleveland, OH 44106Case Comprehensive Cancer Center; Department of Pathology; Department of Medicine, Tuberculosis Research Institute and Division of Infectious Diseases; and Department of Chemistry, Case Western Reserve University, Cleveland, OH 44106.

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2,5-DHBA supplementation confers enhanced sensitivity to E. coli infection. (A) Schematic representation for derivation of 13C-labeled 2,5-DHBA from 13C salicylic acid. (B) Kinetic analysis of 13C-2,5-DHBA clearance from mice. Mice were injected intraperitoneally with 13C-2,5-DHBA and its elimination from the body was assessed by subjecting urine samples to GC-MS analysis. Data are representative of two independent experiments, each with three mice per group. (C) Quantification of 2,5-DHBA levels in urine samples from control and bdh2- mice. 2,5-DHBA levels were quantitated by comparing to known amounts of 2,5-DHBA. Results show pooled data from two independent experiments, each with at least three mice per group. Statistical analysis by two-tailed unpaired Student’s t test: ***, P < 0.001. (D) Serum transferrin saturation in 8-wk old female C57BL/6 (WT) mice injected with 2,5-DHBA. Results show pooled data from two independent experiments, each with at least three mice per group. Statistical analysis by two-tailed unpaired Student’s t test: ns, not significant. (E) Iron levels were measured in nonhemolyzed serum of control and 2,5-DHBA–injected mice. Symbols represent individual mice. Bars represent the mean values. Results show pooled data from two independent experiments, each with at least three mice per group. Statistical analysis by two-tailed unpaired t test: ns, not significant. (F–H) Survival curve comparing WT and bdh2- mice supplemented with 2,5-DHBA or iron or 2,5-DHBA + iron and challenged with 2 × 108 CFU of E. coli H9049 strain. Data are representative of two independent experiments with 4–14 mice per group. Statistical analysis by log-rank test: *, P < 0.05. (I) 2,5-DHBA supplementation augments bacterial growth in mice. WT and bdh2- mice were infected with 0.6 × 108 CFU of E. coli H9049 strain and bacterial loads in blood and parenchymal tissues after infection were determined. Symbols represent individual mice. Bars represent the mean CFU. Data are representative of two independent experiments, each with at least 5 mice per group. Statistical analysis by two-tailed unpaired Student’s t test with Welch’s correction: *, P < 0.05; **, P < 0.01.
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fig4: 2,5-DHBA supplementation confers enhanced sensitivity to E. coli infection. (A) Schematic representation for derivation of 13C-labeled 2,5-DHBA from 13C salicylic acid. (B) Kinetic analysis of 13C-2,5-DHBA clearance from mice. Mice were injected intraperitoneally with 13C-2,5-DHBA and its elimination from the body was assessed by subjecting urine samples to GC-MS analysis. Data are representative of two independent experiments, each with three mice per group. (C) Quantification of 2,5-DHBA levels in urine samples from control and bdh2- mice. 2,5-DHBA levels were quantitated by comparing to known amounts of 2,5-DHBA. Results show pooled data from two independent experiments, each with at least three mice per group. Statistical analysis by two-tailed unpaired Student’s t test: ***, P < 0.001. (D) Serum transferrin saturation in 8-wk old female C57BL/6 (WT) mice injected with 2,5-DHBA. Results show pooled data from two independent experiments, each with at least three mice per group. Statistical analysis by two-tailed unpaired Student’s t test: ns, not significant. (E) Iron levels were measured in nonhemolyzed serum of control and 2,5-DHBA–injected mice. Symbols represent individual mice. Bars represent the mean values. Results show pooled data from two independent experiments, each with at least three mice per group. Statistical analysis by two-tailed unpaired t test: ns, not significant. (F–H) Survival curve comparing WT and bdh2- mice supplemented with 2,5-DHBA or iron or 2,5-DHBA + iron and challenged with 2 × 108 CFU of E. coli H9049 strain. Data are representative of two independent experiments with 4–14 mice per group. Statistical analysis by log-rank test: *, P < 0.05. (I) 2,5-DHBA supplementation augments bacterial growth in mice. WT and bdh2- mice were infected with 0.6 × 108 CFU of E. coli H9049 strain and bacterial loads in blood and parenchymal tissues after infection were determined. Symbols represent individual mice. Bars represent the mean CFU. Data are representative of two independent experiments, each with at least 5 mice per group. Statistical analysis by two-tailed unpaired Student’s t test with Welch’s correction: *, P < 0.05; **, P < 0.01.

Mentions: To determine whether exogenous supplementation with 2,5-DHBA alters the course of E. coli infection in bdh2- mice, we first assessed the kinetics of 2,5-DHBA absorption, distribution, and clearance using 13C-labeled 2,5-DHBA and mass spectrometry. Fig. 4 A shows the derivation of 13C 2,5-DHBA from 13C salicylic acid. Proton (1H) and 13C NMR analyses were used to assess the purity of the synthesized compound. We injected 13C-2,5-DHBA at 20 µg/kg i.p. and studied the kinetics of clearance by analyzing urine samples collected at 4, 8, 12, 24, 48, 72, and 96 h after injection. By 12 h after injection, half of the initial dose of 13C-2,5-DHBA was cleared from the plasma (Fig. 4 B).


Regulation of mammalian siderophore 2,5-DHBA in the innate immune response to infection.

Liu Z, Reba S, Chen WD, Porwal SK, Boom WH, Petersen RB, Rojas R, Viswanathan R, Devireddy L - J. Exp. Med. (2014)

2,5-DHBA supplementation confers enhanced sensitivity to E. coli infection. (A) Schematic representation for derivation of 13C-labeled 2,5-DHBA from 13C salicylic acid. (B) Kinetic analysis of 13C-2,5-DHBA clearance from mice. Mice were injected intraperitoneally with 13C-2,5-DHBA and its elimination from the body was assessed by subjecting urine samples to GC-MS analysis. Data are representative of two independent experiments, each with three mice per group. (C) Quantification of 2,5-DHBA levels in urine samples from control and bdh2- mice. 2,5-DHBA levels were quantitated by comparing to known amounts of 2,5-DHBA. Results show pooled data from two independent experiments, each with at least three mice per group. Statistical analysis by two-tailed unpaired Student’s t test: ***, P < 0.001. (D) Serum transferrin saturation in 8-wk old female C57BL/6 (WT) mice injected with 2,5-DHBA. Results show pooled data from two independent experiments, each with at least three mice per group. Statistical analysis by two-tailed unpaired Student’s t test: ns, not significant. (E) Iron levels were measured in nonhemolyzed serum of control and 2,5-DHBA–injected mice. Symbols represent individual mice. Bars represent the mean values. Results show pooled data from two independent experiments, each with at least three mice per group. Statistical analysis by two-tailed unpaired t test: ns, not significant. (F–H) Survival curve comparing WT and bdh2- mice supplemented with 2,5-DHBA or iron or 2,5-DHBA + iron and challenged with 2 × 108 CFU of E. coli H9049 strain. Data are representative of two independent experiments with 4–14 mice per group. Statistical analysis by log-rank test: *, P < 0.05. (I) 2,5-DHBA supplementation augments bacterial growth in mice. WT and bdh2- mice were infected with 0.6 × 108 CFU of E. coli H9049 strain and bacterial loads in blood and parenchymal tissues after infection were determined. Symbols represent individual mice. Bars represent the mean CFU. Data are representative of two independent experiments, each with at least 5 mice per group. Statistical analysis by two-tailed unpaired Student’s t test with Welch’s correction: *, P < 0.05; **, P < 0.01.
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fig4: 2,5-DHBA supplementation confers enhanced sensitivity to E. coli infection. (A) Schematic representation for derivation of 13C-labeled 2,5-DHBA from 13C salicylic acid. (B) Kinetic analysis of 13C-2,5-DHBA clearance from mice. Mice were injected intraperitoneally with 13C-2,5-DHBA and its elimination from the body was assessed by subjecting urine samples to GC-MS analysis. Data are representative of two independent experiments, each with three mice per group. (C) Quantification of 2,5-DHBA levels in urine samples from control and bdh2- mice. 2,5-DHBA levels were quantitated by comparing to known amounts of 2,5-DHBA. Results show pooled data from two independent experiments, each with at least three mice per group. Statistical analysis by two-tailed unpaired Student’s t test: ***, P < 0.001. (D) Serum transferrin saturation in 8-wk old female C57BL/6 (WT) mice injected with 2,5-DHBA. Results show pooled data from two independent experiments, each with at least three mice per group. Statistical analysis by two-tailed unpaired Student’s t test: ns, not significant. (E) Iron levels were measured in nonhemolyzed serum of control and 2,5-DHBA–injected mice. Symbols represent individual mice. Bars represent the mean values. Results show pooled data from two independent experiments, each with at least three mice per group. Statistical analysis by two-tailed unpaired t test: ns, not significant. (F–H) Survival curve comparing WT and bdh2- mice supplemented with 2,5-DHBA or iron or 2,5-DHBA + iron and challenged with 2 × 108 CFU of E. coli H9049 strain. Data are representative of two independent experiments with 4–14 mice per group. Statistical analysis by log-rank test: *, P < 0.05. (I) 2,5-DHBA supplementation augments bacterial growth in mice. WT and bdh2- mice were infected with 0.6 × 108 CFU of E. coli H9049 strain and bacterial loads in blood and parenchymal tissues after infection were determined. Symbols represent individual mice. Bars represent the mean CFU. Data are representative of two independent experiments, each with at least 5 mice per group. Statistical analysis by two-tailed unpaired Student’s t test with Welch’s correction: *, P < 0.05; **, P < 0.01.
Mentions: To determine whether exogenous supplementation with 2,5-DHBA alters the course of E. coli infection in bdh2- mice, we first assessed the kinetics of 2,5-DHBA absorption, distribution, and clearance using 13C-labeled 2,5-DHBA and mass spectrometry. Fig. 4 A shows the derivation of 13C 2,5-DHBA from 13C salicylic acid. Proton (1H) and 13C NMR analyses were used to assess the purity of the synthesized compound. We injected 13C-2,5-DHBA at 20 µg/kg i.p. and studied the kinetics of clearance by analyzing urine samples collected at 4, 8, 12, 24, 48, 72, and 96 h after injection. By 12 h after injection, half of the initial dose of 13C-2,5-DHBA was cleared from the plasma (Fig. 4 B).

Bottom Line: Pathogens secrete small iron-binding moieties, siderophores, to acquire host iron.In support of this idea, supplementation with mammalian siderophore enhances bacterial growth in vitro.Thus, reciprocal regulation of 24p3 and mammalian siderophore is a protective mechanism limiting microbial access to iron.

View Article: PubMed Central - HTML - PubMed

Affiliation: Case Comprehensive Cancer Center; Department of Pathology; Department of Medicine, Tuberculosis Research Institute and Division of Infectious Diseases; and Department of Chemistry, Case Western Reserve University, Cleveland, OH 44106Case Comprehensive Cancer Center; Department of Pathology; Department of Medicine, Tuberculosis Research Institute and Division of Infectious Diseases; and Department of Chemistry, Case Western Reserve University, Cleveland, OH 44106.

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Related in: MedlinePlus