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Bacterial Infections

Bacillus anthracis Edema Toxin Increases Fractional Free Water and Sodium Reabsorption in an Isolated Perfused Rat Kidney Model

Dharmvir S. Jaswal, Xizhong Cui, Parizad Torabi-Parizi, Lernik Ohanjanian, Hannish Sampath-Kumar, Yvonne Fitz, Yan Li, Wanying Xu, Peter Q. Eichacker
Steven R. Blanke, Editor
Dharmvir S. Jaswal
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Xizhong Cui
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Parizad Torabi-Parizi
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Lernik Ohanjanian
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Hannish Sampath-Kumar
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Yvonne Fitz
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Yan Li
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Wanying Xu
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Peter Q. Eichacker
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Steven R. Blanke
Univ. of Illinois Urbana
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DOI: 10.1128/IAI.00264-17
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  • FIG 1
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    FIG 1

    Timeline of experiments. T0, time zero.

  • FIG 2
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    FIG 2

    (A and D) Serial mean (±SEM) urine cAMP concentrations (A) and urine osmolality levels (D) in kidneys challenged with vasopressin or the diluent control alone and perfused under constant flow. (B, C, and E) Serial mean changes from the baseline (±SEM) for fractional free water reabsorption (B), the urine flow rate (C), and fractional sodium reabsorption (E). The numbers of kidneys employed in these experiments and baseline data for parameters shown as serial changes are noted in Tables 1 and 2, respectively.

  • FIG 3
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    FIG 3

    (A and D) Serial mean (±SEM) urine cAMP concentrations (A) and urine osmolality levels (D) in kidneys challenged with ET or PA alone and perfused under constant flow. (B, C, and E) Serial mean changes from the baseline (±SEM) for fractional free water reabsorption (B), the urine flow rate (C), and fractional sodium reabsorption (E). The numbers of kidneys employed in these experiments and baseline data for parameters shown as serial changes are noted in Tables 1 and 2, respectively.

  • FIG 4
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    FIG 4

    (A and D) Serial mean (±SEM) urine cAMP concentrations (A) and urine osmolality levels (D) in kidneys challenged with ET or PA alone and perfused under constant pressure. (B, C, and E) Serial mean changes from the baseline (±SEM) for fractional free water reabsorption (B), the urine flow rate (C) and fractional sodium reabsorption (E). The numbers of kidneys employed in these experiments and baseline data for parameters shown as serial changes are noted in Tables 1 and 2, respectively.

  • FIG 5
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    FIG 5

    Serial GFRs for experiments with kidneys challenged with ET or PA alone and perfused under constant flow (A) or constant pressure (B). The numbers of kidneys employed in these experiments and baseline data for parameters shown as serial changes are noted in Tables 1 and 2, respectively. * and ** denote levels of significance for overall effects between study groups and for time interactions between groups, respectively.

  • FIG 6
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    FIG 6

    (A and D) Serial mean (±SEM) urine cAMP concentrations (A) and urine osmolality levels (D) in kidneys challenged with ET, treated with raxibacumab or a nonspecific control MAb, and perfused under constant flow. (B, C, and E) Serial mean changes from the baseline (±SEM) for fractional free water reabsorption (B), the urine flow rate (C), and fractional sodium reabsorption (E). The numbers of kidneys employed in these experiments and baseline data for parameters shown as serial changes are noted in Tables 1 and 2, respectively.

  • FIG 7
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    FIG 7

    (A and D) Serial mean (±SEM) urine cAMP concentrations (A) and urine osmolality levels (D) in kidneys challenged with ET, treated with adefovir or a diluent, and perfused under constant flow. (B, C, and E) Serial mean changes from the baseline (±SEM) for fractional free water reabsorption (B), the urine flow rate (C), and fractional sodium reabsorption (E). The numbers of kidneys employed in these experiments and baseline data for parameters shown as serial changes are noted in Tables 1 and 2, respectively.

  • FIG 8
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    FIG 8

    Sample photomicrographs (A, C, E, G, I, and K) and AQP2 fluorescence intensity normalized to the maximum intensity for each image plotted along a 10-μm line crossing tubular cells from extraluminal to intraluminal regions in five selected tubules for each of the six kidneys (B, D, F, H, J, and L). In the photomicrographs, AQP2 staining is shown in pink, while nuclear staining and autofluorescence are shown in gray. Shown are photomicrographs and fluorescence intensity data for one kidney each challenged with AVP (A and B) or HEPES (AVP diluent) (C and D) and for two kidneys each challenged with edema toxin (ET for kidney 1 [ET 1] [E and F] and ET 2 [G and H]) or protective antigen alone (PA for kidney 1 [PA 1] [I and J] and PA 2 [K and L]). While the fluorescence intensity was dispersed widely along the plotted lines across tubules from kidneys challenged with HEPES and PA, this intensity tended to be localized over a short distance for tubules from AVP- and ET-challenged kidneys. As presented in Results, consistent with this narrow localization with ET but not PA, free AQP2 levels were significantly higher in urine from ET-perfused kidneys than in urine from PA-perfused kidneys at 90 and 120 min of perfusion (5.52 ± 1.06 ng/ml [n = 14 kidneys] versus 1.51 ± 0.44 ng/ml [n = 12] [means ± SEM, averaged over the two time points]; P = 0.0001).

Tables

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  • TABLE 1

    Summary of dosages of challenges and treatments and numbers of kidneys employed in the isolated perfused kidney experiments

    ExptMethod of perfusionChallengeTreatmentNo. of kidneys tested
    TypeDoseaDrugDose
    Effect of challenge with vasopressin (AVP)Constant flowAVP0.025 μM 3
    Diluent0 5
    Constant flowAVP0.05 μM 6
    Diluent0 7
    Constant flowAVP0.1 μM 5
    Diluent0 μg/ml 5
    Effect of ET challenge with constant flowConstant flowPA2.0 μg/ml 7
    ET1.0 μg/ml 7
    Constant flowPA4.0 μg/ml 7
    ET2.0 μg/ml 6
    Effect of ET challenge with constant pressureConstant pressurePA1.0 μg/ml 9
    ET0.5 μg/ml 9
    Constant pressurePA2.0 μg/ml 6
    ET1.0 μg/ml 6
    Effect of raxibacumab treatment with ET challengeConstant flowET1.0 μg/mlRaxibacumab10×b8
    ET1.0 μg/mlControl06
    Constant flowET1.0 μg/mlRaxibacumab50×3
    ET1.0 μg/mlControl03
    Effect of adefovir treatment with ET challengeConstant flowET1.0 μg/mlAdefovir20 μM7
    ET1.0 μg/mlControl05
    Constant flowET2.0 μg/mlAdefovir20 μM4
    ET2.0 μg/mlControl04
    • ↵a ET consists of edema factor and PA combined in a weight ratio of 1:2, and the dose of ET shown denotes the amount of EF employed.

    • ↵b Molar amount of either raxibacumab or a nonspecific (control) MAb compared to PA.

  • TABLE 2

    Baseline measurementsa

    ParameterMean value ± SD
    Effect of challenge with vasopressin (AVP)Effect of ET challenge with constant pressureEffect of Raxi with ET challengeEffect of adefovir with ET challengeEffect of ET challenge with constant flow
    DiluentAVPPAETET + NS-AbET + RaxiET + diluentET + adefovirPAET
    Flow (ml/min)32.8 ± 1.031.4 ± 0.929.3 ± 1.327.5 ± 0.833.1 ± 1.736.1 ± 1.630.8 ± 0.730.8 ± 0.929.5 ± 0.930.4 ± 0.6
    Perfusion pressure (mm Hg)101.8 ± 1.4100.7 ± 0.999.9 ± 0.9100.8 ± 0.898.8 ± 0.597.7 ± 1.0100.9 ± 1.0101.8 ± 1.0101.1 ± 1.6100.9 ± 1.6
    GFR (ml/min)0.83 ± 0.030.88 ± 0.050.71 ± 0.050.77 ± 0.060.77 ± 0.040.83 ± 0.040.75 ± 0.030.79 ± 0.040.75 ± 0.030.77 ± 0.03
    Fractional free water reabsorption0.81 ± 0.010.81 ± 0.010.88 ± 0.010.88 ± 0.010.83 ± 0.010.84 ± 0.010.84 ± 0.010.84 ± 0.010.84 ± 0.010.83 ± 0.02
    Urine output (μl/min)157.2 ± 10.9168.1 ± 11.186.6 ± 8.495.4 ± 10.5132.8 ± 13.9135.7 ± 11.0123.5 ± 7.8125.7 ± 9.6126 ± 15130 ± 15
    Fractional sodium reabsorption0.82 ± 0.010.81 ± 0.010.88 ± 0.010.88 ± 0.010.83 ± 0.010.84 ± 0.010.84 ± 0.010.85 ± 0.010.84 ± 0.020.84 ± 0.02
    • ↵a Raxi, raxibacumab; NS-Ab, nonspecific antibody.

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Bacillus anthracis Edema Toxin Increases Fractional Free Water and Sodium Reabsorption in an Isolated Perfused Rat Kidney Model
Dharmvir S. Jaswal, Xizhong Cui, Parizad Torabi-Parizi, Lernik Ohanjanian, Hannish Sampath-Kumar, Yvonne Fitz, Yan Li, Wanying Xu, Peter Q. Eichacker
Infection and Immunity Jun 2017, 85 (7) e00264-17; DOI: 10.1128/IAI.00264-17

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Bacillus anthracis Edema Toxin Increases Fractional Free Water and Sodium Reabsorption in an Isolated Perfused Rat Kidney Model
Dharmvir S. Jaswal, Xizhong Cui, Parizad Torabi-Parizi, Lernik Ohanjanian, Hannish Sampath-Kumar, Yvonne Fitz, Yan Li, Wanying Xu, Peter Q. Eichacker
Infection and Immunity Jun 2017, 85 (7) e00264-17; DOI: 10.1128/IAI.00264-17
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    • ABSTRACT
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KEYWORDS

Antigens, Bacterial
bacterial toxins
kidney
Sodium
Water
anthrax
edema toxin
kidney
water reabsorption

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