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Overexpression of IFN-Induced Protein 10 and Its Receptor CXCR3 in Myasthenia Gravis

Tali Feferman, Prasanta K. Maiti, Sonia Berrih-Aknin, Jacky Bismuth, Jocelyne Bidault, Sara Fuchs and Miriam C. Souroujon
J Immunol May 1, 2005, 174 (9) 5324-5331; DOI: https://doi.org/10.4049/jimmunol.174.9.5324
Tali Feferman
*Department of Immunology, The Weizmann Institute of Science, Rehovot, Israel;
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Prasanta K. Maiti
*Department of Immunology, The Weizmann Institute of Science, Rehovot, Israel;
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Sonia Berrih-Aknin
*Department of Immunology, The Weizmann Institute of Science, Rehovot, Israel;
†Centre National de la Recherche Scientifique, Unité Mixte de Recherche 8078, Unité Paris Sud, Institut Paris Sud Cytokines, Hospital Marie Lannelongue, Le Plessis Robinson, France; and
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Jacky Bismuth
†Centre National de la Recherche Scientifique, Unité Mixte de Recherche 8078, Unité Paris Sud, Institut Paris Sud Cytokines, Hospital Marie Lannelongue, Le Plessis Robinson, France; and
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Jocelyne Bidault
†Centre National de la Recherche Scientifique, Unité Mixte de Recherche 8078, Unité Paris Sud, Institut Paris Sud Cytokines, Hospital Marie Lannelongue, Le Plessis Robinson, France; and
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Sara Fuchs
*Department of Immunology, The Weizmann Institute of Science, Rehovot, Israel;
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Miriam C. Souroujon
*Department of Immunology, The Weizmann Institute of Science, Rehovot, Israel;
‡Open University of Israel, Raanana, Israel
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  • FIGURE 1.
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    FIGURE 1.

    mRNA expression of chemokines and cytokines in LNC and muscles of EAMG and healthy rats. LNC and muscle samples from at least four individual rats from each experimental group were harvested in each experiment. Myasthenic rats were killed when they reached a clinical score of 2; for each myasthenic rat, a control rat was killed at the same time point. mRNA expression levels of cytokines and chemokines in LNC (A) and muscles (B) from EAMG and control CFA healthy rats were determined by Q-RT-PCR, and data are presented as the relative expression value for every EAMG group compared with its control, which was assigned a value of 100. β-Actin was used as an inner control for normalization for each cytokine/chemokine. ∗, p < 0.05; ∗∗, p < 0.005. The data shown are representative of five independent experiments

  • FIGURE 2.
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    FIGURE 2.

    Cytokine and chemokine mRNA expression after successful treatment of EAMG by Rα1–205. LNC and muscle samples from at least four individual rats from each experimental group were harvested in each experiment. mRNA expression levels of cytokines in LNC (A) and muscles (B) from rats in which treatment with Rα1–205 started during the acute or chronic phase of EAMG were determined by Q-RT-PCR 8 wk after disease induction, and data are presented as the relative expression value for the Rα1–205-treated group compared with OVA-treated controls, which were assigned a value of 100. β-Actin was used for normalization for each cytokine/chemokine. ∗, p < 0.05. The data shown are representative of three independent experiments.

  • FIGURE 3.
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    FIGURE 3.

    mRNA and protein expression levels of ICAM-1 and LFA-1 in LNC and splenocytes of EAMG and healthy rats. LNC and splenocytes from EAMG and healthy CFA-injected control rats were harvested when rats reached a clinical score of 2. A, mRNA expression levels of ICAM-1 and LFA-1 from EAMG and control CFA healthy rats were determined by Q-RT-PCR, and data are presented as the relative expression value for every EAMG group compared with its control, which was assigned a value of 100. β-Actin was used as an inner control for normalization for each gene. ∗, p < 0.05 (n = 4). The data shown are representative of five experiments. B, Protein levels of ICAM-1 and LFA-1 were determined in splenocytes from EAMG and healthy CFA-injected control rats (two or three rats in each group) that were subjected to FACS analysis with PE-conjugated anti-CD4 or anti-CD8 Abs combined with FITC-conjugated anti-ICAM-1 or anti-LFA-1 Abs. Shown is a representative experiment of five independent experiments performed.

  • FIGURE 4.
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    FIGURE 4.

    mRNA expression levels of IP-10 and CXCR3 in the thymus of MG patients and healthy controls. mRNA expression levels of IP-10 and CXCR3 were analyzed by Q-RT-PCR in thymuses of MG patients (n = 5) compared with controls (n = 6). The results are shown as the mean ± SEM. The p values were obtained using the nonparametric Mann-Whitney U test.

  • FIGURE 5.
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    FIGURE 5.

    CXCR3 surface expression in PBMC of MG patients and controls. PBMC from MG patients (n = 6) and age-matched controls (n = 7) were incubated first with anti-CXCR3 Abs, followed by FITC-conjugated goat anti-mouse Ig. The cells were further incubated with mouse IgG1 to saturate the free sites of the FITC-goat anti-mouse Ig. After washings, the cells were incubated with one of the following mAbs: anti-CD19-R-PE, anti-CD4-R-PE-Cy5, or anti-CD8-R-PE. The results are shown as the mean ± SEM. The p values were obtained using the nonparametric Mann-Whitney U test.

  • FIGURE 6.
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    FIGURE 6.

    mRNA expression levels of IP-10 and CXCR3 in muscles of MG patients. mRNA expression levels of IP-10 and CXCR3 were analyzed in muscles of MG patients compared with controls. The results are shown as the mean ± SEM of 17 MG patients and four controls. The inset shows a positive correlation between the expression levels of IP-10 and CXCR3 in human muscles biopsies. The p values were obtained using the nonparametric Mann-Whitney U test.

  • FIGURE 7.
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    FIGURE 7.

    CXCR3 protein in myotubes of MG patients and controls. Frozen sections from the muscle of an MG patient (A) and a normal control (B) were stained with anti-CXCR3 Abs. The mean fluorescence was quantified on the whole section using Image J software (C). The results are expressed as the mean fluorescence intensity of CXCR3-stained sections of six MG patients and four controls (after deduction of the relevant background fluorescence intensity).

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The Journal of Immunology: 174 (9)
The Journal of Immunology
Vol. 174, Issue 9
1 May 2005
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Overexpression of IFN-Induced Protein 10 and Its Receptor CXCR3 in Myasthenia Gravis
Tali Feferman, Prasanta K. Maiti, Sonia Berrih-Aknin, Jacky Bismuth, Jocelyne Bidault, Sara Fuchs, Miriam C. Souroujon
The Journal of Immunology May 1, 2005, 174 (9) 5324-5331; DOI: 10.4049/jimmunol.174.9.5324

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Overexpression of IFN-Induced Protein 10 and Its Receptor CXCR3 in Myasthenia Gravis
Tali Feferman, Prasanta K. Maiti, Sonia Berrih-Aknin, Jacky Bismuth, Jocelyne Bidault, Sara Fuchs, Miriam C. Souroujon
The Journal of Immunology May 1, 2005, 174 (9) 5324-5331; DOI: 10.4049/jimmunol.174.9.5324
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