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Bacterial Infections

H2S, a Bacterial Defense Mechanism against the Host Immune Response

Tracy Toliver-Kinsky, Weihua Cui, Gabor Törö, Seung-Jin Lee, Konstantin Shatalin, Evgeny Nudler, Csaba Szabo
Nancy E. Freitag, Editor
Tracy Toliver-Kinsky
aDepartment of Anesthesiology, University of Texas Medical Branch, Galveston, Texas, USA
bShriners Hospitals for Children, Galveston, Texas, USA
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Weihua Cui
aDepartment of Anesthesiology, University of Texas Medical Branch, Galveston, Texas, USA
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Gabor Törö
aDepartment of Anesthesiology, University of Texas Medical Branch, Galveston, Texas, USA
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Seung-Jin Lee
aDepartment of Anesthesiology, University of Texas Medical Branch, Galveston, Texas, USA
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Konstantin Shatalin
cDepartment of Biochemistry and Molecular Pharmacology, Howard Hughes Medical Institute, New York University School of Medicine, New York, New York, USA
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Evgeny Nudler
cDepartment of Biochemistry and Molecular Pharmacology, Howard Hughes Medical Institute, New York University School of Medicine, New York, New York, USA
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Csaba Szabo
aDepartment of Anesthesiology, University of Texas Medical Branch, Galveston, Texas, USA
bShriners Hospitals for Children, Galveston, Texas, USA
dChair of Pharmacology, Department of Medicine, University of Fribourg, Fribourg, Switzerland
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Nancy E. Freitag
University of Illinois at Chicago
Roles: Editor
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DOI: 10.1128/IAI.00272-18
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    FIG 1

    E. coli and S. aureus were cultured with increasing concentrations of up to 1 mM GYY4137 prior to inoculation of mouse leukocyte cultures. Graphs show percentages of inoculum CFU killed after coculture with leukocytes. *, significantly different from 0 mM (n = 4 to 5 replicates per group).

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    FIG 2

    (A) Image showing brown lead sulfide staining produced by reaction of lead acetate with H2S produced by E. coli cultured with or without the 3-MST inhibitor. The graph on the left shows the activity of bacterial 3MST in the presence of increasing concentrations of the 3-MST inhibitor, expressed as a percentage of 3MST activity in the control (no inhibitor) (n = 3 replicates/group). The graph on the right shows in vitro elimination of E. coli by leukocytes after 45 min. WT E. coli bacteria were cultured in the presence of the 3-MST inhibitor prior to inoculation of leukocytes. *, significantly different from 0 μM; #, significantly different from 1 μM (n = 5/group). (B) Brown lead sulfide stain produced by reaction of lead acetate with H2S from S. aureus cultured with or without increasing concentrations of AOAA or PAG, in the presence (+Cys) or absence (−Cys) of 200 µM cysteine supplementation. Graphs show in vitro elimination of S. aureus by leukocytes after 60 min. WT S. aureus bacteria were cultured with inhibitors prior to inoculation of leukocytes. *, significantly different from 0 mM; #, significantly different from 3 mM (n = 5/group).

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    (A) Images showing brown lead sulfide staining produced by reaction of lead acetate with H2S produced by bacterial cultures. WT, wild-type E. coli; ΔsseA, sseA-deficient E. coli. (Left) Bacterial elimination by leukocytes after 45 min in vitro. *, significantly different from the WT (n = 6/group). (Right) Densities of E. coli WT and ΔsseA bacteria in liquid cultures over 5 h, measured by the OD600, and viability in cell culture medium, measured as CFU per milliliter (n = 3/group). (B) E. coli ΔsseA bacteria were cultured in the presence of GYY4137 prior to inoculation of mouse leukocytes. The graph shows bacterial elimination after 45 min of coculture. *, significantly different from 0 mM (n = 4 per group). (C, left) Total killing of E. coli bacteria by RAW 264.7 macrophages after 1 and 2 h in vitro. *, significantly different from all other groups (n = 7/group). (Right) Levels (CFU per milliliter) of viable intracellular E. coli bacteria recovered from RAW 264.7 cells. *, significantly different from the ΔsseA group at the corresponding time point (n = 6/group and time point).

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    (A) Images showing brown lead sulfide staining produced by reaction of lead acetate with H2S produced by bacteria grown with or without cysteine (Cys) (200 μM) supplementation. WT, wild-type S. aureus; Δcbs Δcse, S. aureus lacking the cbs and cse genes. The graph on the left shows bacterial clearance by leukocytes after 60 min in vitro. *, significantly different from the WT (n = 6/group). The graphs on the right show densities of S. aureus WT and Δcbs Δcse bacteria in liquid cultures over 5 h, measured by the OD600, and viability in cell culture medium, measured as CFU per milliliter (n = 3/group). (B) S. aureus Δcbs Δcse bacteria were cultured in the presence of GYY4137 prior to inoculation of mouse leukocytes. The graph shows the percentage of the inoculum cleared after coculture with leukocytes. *, significantly different from 0 and 0.1 mM (n = 5 per group).

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    FIG 5

    (A, left) Bar graphs showing bacterial counts in spleen and IL-6 levels in plasma 16 h after i.p. injection of mice with 1 × 108 CFU E. coli. *, significantly different from the WT (n = 5 mice/group). (Right) Densities of E. coli WT and ΔsseA bacteria in liquid cultures, measured by the OD600 over 16 h. (B, left) Bar graphs showing bacterial counts in spleens and plasma IL-6 levels in mice 16 h after i.p. injection of 5 × 108 CFU S. aureus. *, significantly different from the WT (n = 5/group). (Right) Densities of S. aureus WT and Δcbs Δcse bacteria in liquid cultures, measured by the OD600 over 16 h. (C, left) Graph showing bacterial counts in spleens of burned mice 16 h after i.p. injection of mice with 1 × 108 CFU E. coli. *, significantly different from the WT (n = 5 mice/group). (Right) Graphs showing bacterial counts in burn wounds and spleens of mice 24 h after inoculation of wounds with 1 × 106 CFU S. aureus. *, significantly different from the WT (n = 3 to 4/group).

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H2S, a Bacterial Defense Mechanism against the Host Immune Response
Tracy Toliver-Kinsky, Weihua Cui, Gabor Törö, Seung-Jin Lee, Konstantin Shatalin, Evgeny Nudler, Csaba Szabo
Infection and Immunity Dec 2018, 87 (1) e00272-18; DOI: 10.1128/IAI.00272-18

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H2S, a Bacterial Defense Mechanism against the Host Immune Response
Tracy Toliver-Kinsky, Weihua Cui, Gabor Törö, Seung-Jin Lee, Konstantin Shatalin, Evgeny Nudler, Csaba Szabo
Infection and Immunity Dec 2018, 87 (1) e00272-18; DOI: 10.1128/IAI.00272-18
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KEYWORDS

antibiotic resistance
burn
hydrogen sulfide
opportunistic infections

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