13 research outputs found

    Motor coordination in Theiler’s murine encephalomyelitis virus (TMEV)-infected SJL mice.

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    <p>(A) <i>Experiment I</i>: RotaRod<sup>®</sup> performance test after early application of the IL-10 receptor blocking antibody (IL-10R Ab) at 0, 7, 14 and 21 dpi (arrows) revealed motor coordination deficits in TMEV-infected animals compared to mock-infected mice at 42 dpi. However, no differences were determined between IL-10R Ab- and isotype-treated animals. white box = TMEV-infected mice without IL-10R Ab treatment (group “isotype<sub>early</sub>/TMEV”), grey box = TMEV-infected mice with IL-10R treatment (group “IL10R↓<sub>early</sub>/TMEV”), black box = mock-infected mice with IL-10R treatment (group “IL10R↓<sub>early</sub>/mock”). (B) <i>Experiment II</i>: application of IL-10R Ab at 35 and 42 dpi (arrows) caused also deterioration of motor coordination starting at 42 dpi. However, no differences were observed between IL-10R Ab- and isotype-treated animals (groups “IL-10R↓<sub>late</sub>/TMEV” and “isotype<sub>late</sub>/TMEV”). white box = TMEV-infected mice without IL-10R Ab treatment (group “isotype<sub>late</sub>/TMEV”), grey box = TMEV-infected mice with IL-10R treatment (group “IL10R↓<sub>late</sub>/TMEV”), black box = mock-infected mice with IL-10R treatment (group “IL10R↓<sub>late</sub>/mock”). Box and whisker plots display median, minimum and maximum values as well as upper and lower quartiles, 5 animals used at all investigated time points, Wilcoxon rank-sum tests, *  =  p < 0.05.</p

    Effect of IL-10 receptor (IL-10R) blockade upon leukomyelitis in Theiler’s murine encephalomyelitis virus (TMEV)-infected SJL mice.

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    <p>(A) TMEV-infection and concurrent intraperitoneal application of IL-10R antibody lead to an increase of CD3<sup>+</sup> T cells in the spinal cord at 49 days post infection (dpi) compared to (B) TMEV-infected control animals. A,B: immunohistochemistry, bar = 20 μm. (C) Statistical analyses revealed a significant increase of CD3<sup>+</sup> T cells in TMEV-infected mice with IL-10R blockade compared to isotype-treated animals at 49 dpi. Almost no CD3<sup>+</sup> T cells were present in spinal cord cross sections of mock-infected animals. (D) Simultaneously, IL-6 mRNA expression significantly decreased in TMEV-infected animals following IL-10R blockade. white box = TMEV-infected mice without IL-10R Ab treatment (group “isotype<sub>late</sub>/TMEV”), grey box = TMEV-infected mice with IL-10R treatment (group “IL10R↓<sub>late</sub>/TMEV”), black box = mock-infected mice with IL-10R treatment (group “IL10R↓<sub>late</sub>/mock”). Box and whisker plots display median, minimum and maximum values as well as upper and lower quartiles, 5 animals used in all groups, Wilcoxon rank-sum tests, *  =  p < 0.05.</p

    Leukocyte subsets in the colon of interleukin-10 receptor (IL-10R) neutralized SJL mice.

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    <p>(A) Application of IL-10R antibody (Ab) leads to increased numbers of CD3<sup>+</sup> T cells in the colon at all investigated time points. (B). An increase of CD45R<sup>+</sup> B cells can be observed at day 21 after the first Ab application. (C) Foxp3<sup>+</sup> regulatory T cells and (D) Iba-1<sup>+</sup> macrophages are elevated at 7, 14, and 21 dpi. grey box with vertical lines = control animals (group “isotype”), white box with vertical lines = IL-10R blocked animals (group “IL-10R↓”). Box and whisker plot display median, minimum and maximum values as well as upper and lower quartiles, 5 animals used at all investigated time points, Wilcoxon rank-sum tests, *  =  p < 0.05.</p

    Clinical effects of Theiler’s murine encephalomyelitis virus (TMEV)-infection in SJL mice following IL-10R blockade.

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    <p>(A) <i>Experiment I</i>: TMEV-infection causes worsening of systemic clinical signs (increased clinical scores) at 22 dpi in SJL mice with early anti-IL-10 receptor antibody (IL-10R Ab) treatment compared to non-infected mice following anti-IL-10R Ab treatment (<b>red asterisks</b>) and TMEV-infected mice without anti-IL-10R Ab treatment (<b>black asterisks</b>), suggestive of a triggering effect of virus infection upon IL-10R deficiency-mediated systemic signs. (B) <i>Experiment II</i>: Ab treatment during the late infection phase leads to similar clinical scores between TMEV-infected mice with and without IL-10R blockade. 5 animals used in all three groups and at all investigated time points, Wilcoxon rank-sum tests, arrows = administration of IL-10R Ab or isotype control, respectively, red asterisks = significant differences (p < 0.05) between TMEV-infected mice with and without IL-10R blockade, black asterisks = significant differences (p < 0.05) between IL-10R blocked mice with and without TMEV-infection, green asterisks significant differences (p < 0.05) between TMEV-infected mice and IL-10R blocked mice.</p

    Effects of acute Theiler’s murine encephalomyelitis virus (TMEV)-infection (<i>experiment I</i>) upon cytokine expression and phenotypical changes in spleens of interleukin-10 receptor (IL-10R) blocked SJL mice.

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    <p>(A-I) Significantly elevated mRNA levels of IL-1α, IL-2, IL-4, IL-5, IL-6, TNF, IFN-γ, TGF-β and IL-10 in spleens of infected mice (group “IL-10R↓<sub>early</sub>/TMEV”) compared to non-infected animals following IL-10R antibody (Ab) treatment (group “IL10R↓<sub>early</sub>/mock”). (J) Flow cytometry revealed a relative increase of CD19<sup>+</sup> B cells and a simultaneous decrease of CD4<sup>+</sup> T cells (K) in the spleen of infected mice following IL-10R blockade at 14 dpi. (L) Additionally, the relative numbers of CD4<sup>+</sup> Foxp3<sup>+</sup> Treg in the spleen were increased following TMEV-infection and IL-10R blockade. Moreover, gMFI of CD69 (M) and CD44 (N) gated on CD4<sup>+</sup> cells and gMFI of CD69 (O) and CD44 (P) gated on CD8<sup>+</sup> cells were increased at 14 dpi in infected mice following IL-10R Ab treatment. For gating strategy see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0161883#pone.0161883.s001" target="_blank">S1 Fig</a>. IL-10R Ab treated animals without TMEV-infection (group “IL10R↓<sub>early</sub>/mock”). IL-10R Ab treated mice with additional TMEV-infection (group “IL-10R↓<sub>early</sub>/TMEV”). Box and whisker plots display median, minimum and maximum values as well as upper and lower quartiles, 5 animals used in both groups and at all investigated time points, Wilcoxon rank-sum tests, *  =  p < 0.05.</p

    Experimental design.

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    <p>(A) <i>Experiment I</i>: animals infected with Theiler’s murine encephalomyelitis virus (TMEV) received IL-10 receptor blocking antibodies (IL-10R Ab) once weekly at day 0, 7, 14 and 21 of the experiment (grey box). Necropsy (†) was performed in groups of five animals at 7, 14, 22 and 42 days post infection (dpi). For negative controls a second group received IgG1-specific isotype control instead of IL-10R Ab (*) and a third group was mock-infected only (#) (B) <i>Experiment II</i>: TMEV-infected animals received IL-10R Ab once weekly at 35 and 42 dpi (grey box). Necropsy (†) was performed in groups of five animals at 42 and 49 dpi. For negative controls, a second group received IgG1-specific isotype control instead of IL-10R Ab (*) and a third group was mock-infected only (#) (C) <i>Experiment III</i>: animals received IL-10R Ab once weekly at day 0, 7 and 14 (grey box). Necropsy (†) was performed in groups of five animals at 7, 14 and 21 days after the first Ab application. For negative control a second group received IgG1-specific isotype control instead of IL-10R Ab (*).</p

    Effects of chronic Theiler’s murine encephalomyelitis virus (TMEV)-infection (<i>experiment II</i>) on cytokine expression and phenotypical changes in spleens of interleukin-10 receptor (IL-10R) neutralized SJL mice.

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    <p>(A-H) Significantly elevated mRNA levels of IL-1α, IL-2, IL-4, IL-6, TNF, IFN-γ, TGF-β and IL-10 in spleens of infected mice (group “IL-10R↓<sub>late</sub>/TMEV”) compared to non-infected animals following IL-10R antibody (Ab) treatment (group “IL-10R↓<sub>late</sub>/mock”). (I) Flow cytometry revealed a relative increase of CD8<sup>+</sup> cells in the spleens of TMEV-infected mice with IL-10R Ab treatment at 49 dpi. For gating strategy see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0161883#pone.0161883.s002" target="_blank">S2 Fig</a>. IL-10R Ab treated mice without TMEV-infection (group “IL-10R↓<sub>late</sub>/mock”). IL-10R Ab treated mice with TMEV-infection (group “IL-10R↓<sub>late</sub>/TMEV”). Box and whisker plots display median, minimum and maximum values as well as upper and lower quartiles, 5 animals used in both groups, Wilcoxon rank-sum tests, *  =  p < 0.05.</p

    Theiler’s murine encephalomyelitis virus (TMEV)-infection exacerbates enteric disease following interleukin-10 receptor (IL-10R) blockade.

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    <p>(A) Note changes of colonic mucosa with severe lymphohistiocytic to neutrophilic inflammation (arrow heads), crypt abscess formation (#), and loss of crypt epithelium (arrow). H&E staining, bar = 60 μm (B) Significantly increased severity of colitis at 14 days post infection (dpi) in TMEV-infected mice with IL10R antibody (Ab) treatment (group “IL10R↓<sub>early</sub>/TMEV”) compared to Ab treated animals without infection (group “IL10R↓<sub>early</sub>/mock”). Animals receiving isotype control instead of IL-10R Ab did not show any intestinal inflammation. IL-10R Ab treated SJL mice with TMEV-infection (group “IL10R↓<sub>early</sub>/TMEV”), TMEV-infected mice without IL-10R Ab treatment (group “isotype<sub>early</sub>/TMEV”), IL-10R Ab treated SJL mice without TMEV-infection (group “IL10R↓<sub>early</sub>/mock”). Box and whisker plots display median, minimum and maximum values as well as upper and lower quartiles, 5 animals used in all groups, Wilcoxon rank-sum tests, *  =  p < 0.05.</p

    Enteric disease following interleukin-10 receptor (IL-10R) blockade in non-infected SJL mice.

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    <p>(A) Unformed feces and mucosal edema in an animal at 14 days after onset of IL-10R antibody (Ab) treatment. (B) Colon of a control animal (group “isotype”) at day 21 with normal mucosal architecture. (C) Colon of an animal receiving IL-10R Ab (group “IL-10R↓”) at the same time point. Note infiltration of neutrophilic granulocytes (arrows) and loss of goblet cells (#). B, C: H&E staining, bar = 20 μm. (D) IL-10R blockade causes progressive colitis compared to control animals. grey box with vertical lines = control animals (group “isotype”), white box with vertical lines = IL-10R blocked animals (group “IL-10R↓”). Box and whisker plot display median, minimum and maximum values as well as upper and lower quartiles, 5 animals used at all investigated time points, Wilcoxon rank-sum tests, *  =  p < 0.05.</p
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