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IL-17 Stimulates Granulopoiesis in Mice: Use of an Alternate, Novel Gene Therapy-Derived Method for In Vivo Evaluation of Cytokines

Paul Schwarzenberger, Vincent La Russa, Allan Miller, Peng Ye, Weitao Huang, Arthur Zieske, Steve Nelson, Gregory J. Bagby, David Stoltz, Randall L. Mynatt, Melanie Spriggs and Jay K. Kolls
J Immunol December 1, 1998, 161 (11) 6383-6389;
Paul Schwarzenberger
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Vincent La Russa
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Allan Miller
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Peng Ye
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Weitao Huang
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Arthur Zieske
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Steve Nelson
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Gregory J. Bagby
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David Stoltz
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Randall L. Mynatt
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Melanie Spriggs
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Jay K. Kolls
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  • FIGURE 1.
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    FIGURE 1.

    A, IL-17 pharmacokinetics. Serum obtained from individual mice (n = 4 per group) was analyzed at serial time points for mIL-17 bioactivity as described in Materials and Methods. Specificity of the reaction was confirmed with anti-mIL-17 serum, which in this assay completely inhibited mIL-17 bioactivity. No mIL-17 expression was detected in control animals; therefore, the depicted AdCMVLuc curve is identical with background mIL-17 levels (AdCMVLuc- or PBS-treated animals). Data are expressed as mIL-17 units and represent the mean value of individual animals ± SEM. *, Value at least p < 0.05. B, mIL-17 detection in plasma by Western blot. Plasma from individual Ad-mIL-17-treated animals was analyzed for mIL-17 protein at different time points as outlined in Materials and Methods: day 3 (lane 3), day 7 (lane 4), day 14 (lane 5), and day 21 (lane 6). Lane 7 is plasma from an untreated control animal. Supernatant from Ad-mIL-17-infected 911 producer cells and recombinant E. coli-produced mIL-17 are depicted in lanes 1 and 2, respectively. C, Demonstration of mIL-17 gene transfer to the liver. Presence of the transferred mIL-17 gene in the liver of treated mice was confirmed at serial time points. A specific 477-bp product (pos. = positive control) is visualized in Ad-mIL-17-treated animals over a period of 21 days (days 3, 7, 14, and 21: lanes 5-8), but not in control animals (H2O, PBS treated, negative mouse, and AdCMVLuc-treated mice (days 3, 7, 14, and 21: lanes 1-4)).

  • FIGURE 2.
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    FIGURE 2.

    Overexpression of mIL-17 induces splenomegaly. Spleens of four individual animals per time point were harvested after sacrifice and weighed. Data represent the mean value of individual animals ± SEM. *, Indicates a value of at least p < 0.05.

  • FIGURE 3.
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    FIGURE 3.

    mIL-17 causes leukocytosis. Peripheral blood from four individual animals per group was evaluated at serial time points for WBC (A) and ANC (B). The data represent the mean value of results obtained from individual animals ± SEM. Four separate experiments using the same number of animals showed similar results. *, Indicates a value of at least p < 0.05.

  • FIGURE 4.
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    FIGURE 4.

    IL-17 increases bone marrow colony forming units. Bone marrow colonies were plated from two individual animals per data point as described in Materials and Methods, and CFU-GM and CFU-GEEM (A) and HPPC (B) were enumerated. Data represent the mean value (CFU/105 cells) of individual animals ± SEM. *, Indicates a value of at least p < 0.05. The experiment was repeated twice with similar results.

  • FIGURE 5.
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    FIGURE 5.

    IL-17 increases splenic colony forming units. Splenocyte colonies were plated from two individual animals per data point as described in Materials and Methods, and CFU-GM and CFU-GEMM (A) and HPPC (B) were enumerated. Data represent the mean value (CFU/105 cells) of individual animals ± SEM. *, Indicates a value of at least p < 0.05. The experiment was repeated twice with similar results.

  • FIGURE 6.
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    FIGURE 6.

    IL-17 induces transiently elevated G-CSF serum levels. Serum content of G-CSF was determined in four individual animals per group as outlined in Materials and Methods. No G-CSF was detected in either group on days 7, 14, or 21. Data represent the mean value of individual animals ± SEM. *, Indicates a value of at least p < 0.05. The experiment was repeated once with similar results.

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    Table I.

    mIL-17 increases total spleen cellularitya

    CellDay
    0371421
    Spleen Ad-IL-1710.4 ± 2.112.44 ± 1.918.92b ± 2.218.46 ± 2.4b13.24 ± 2.6
    Spleen Ad-luc9.8 ± 1.98.13 ± 2.312.54 ± 2.313.2 ± 1.812.84 ± 3.2
    BM Ad-IL-171.1 ± 0.31.05 ± 0.231.65 ± 0.192.42 ± 0.462.26 ± 0.33
    BM Ad-luc1.3 ± 0.221.46 ± 0.311.96 ± 0.421.96 ± 0.392.35 ± 0.4
    • a Spleen and bone marrow cells from three individual animals per group were obtained and processed at different time points as outlined in Materials and Methods. Numbers are expressed as cells × 107. The data represent the mean value of results from individual animals ± SEM. Two subsequent experiments using the same number of animals showed similar results.

    • b , Indicates a value of at least p < 0.05.

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The Journal of Immunology
Vol. 161, Issue 11
1 Dec 1998
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IL-17 Stimulates Granulopoiesis in Mice: Use of an Alternate, Novel Gene Therapy-Derived Method for In Vivo Evaluation of Cytokines
Paul Schwarzenberger, Vincent La Russa, Allan Miller, Peng Ye, Weitao Huang, Arthur Zieske, Steve Nelson, Gregory J. Bagby, David Stoltz, Randall L. Mynatt, Melanie Spriggs, Jay K. Kolls
The Journal of Immunology December 1, 1998, 161 (11) 6383-6389;

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IL-17 Stimulates Granulopoiesis in Mice: Use of an Alternate, Novel Gene Therapy-Derived Method for In Vivo Evaluation of Cytokines
Paul Schwarzenberger, Vincent La Russa, Allan Miller, Peng Ye, Weitao Huang, Arthur Zieske, Steve Nelson, Gregory J. Bagby, David Stoltz, Randall L. Mynatt, Melanie Spriggs, Jay K. Kolls
The Journal of Immunology December 1, 1998, 161 (11) 6383-6389;
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