Additional file 1 of Blocking interleukin-23 ameliorates neuromuscular and thymic defects in myasthenia gravis
Creators: name: Villegas, José A.
name: Van Wassenhove, Jérôme
name: Merrheim, Judith
name: Matta, Karen
name: Hamadache, Samy
name: Flaugère, Clémence
name: Pothin, Pauline
name: Truffault, Frédérique
name: Hascoët, Sébastien
name: Santelmo, Nicola
name: Alifano, Marco
name: Berrih-Aknin, Sonia
name: le Panse, Rozen
name: Dragin, Nadine
Publisher: name: figshare
Year: 2023
Subjects: subject: Immunology
subject: FOS: Clinical medicine

description: Additional file 1: Figure S1. Anti-IL-23p19 treatment decreases the activation of SCs and inflammation in EAMG muscle after 2 weeks of treatment. mRNA expression of Pax7 (A), MyoD (B), MyoG (C), Il-6 (D), Il-6r (E), Tgf-β (F) and Il-17a (G) in the Tibialis anterior muscle of CFA and EAMG mice treated with or without anti-IL-23p19 antibody. mRNA analyses were performed in duplicate after 2 weeks treatment. Data were obtained from 2 independent experiments. There were n &gt; 4 mice per group. Each point represents an individual mouse. mRNA expression were determined in duplicate by quantitative RT-PCR. mRNA levels are expressed as arbitrary unit (AU) and normalized to Cypa. P values were obtained with a t-test. P value are δ = 0.06 to 0.05; * &lt; 0.05;**0.005; *** 0.0007. Figure S2. Effect of anti-IL-23p19 treatment on human Th17/Treg cells and antibodies in the spleen and blood of engrafted NSG mice. Analysis by flow cytometry of human T cells in the spleens of NSG-MG mice (A). ELISA analysis of human anti-AChR antibodies in the blood of NSG-MG mice 28 days after thymic engraftment (B). Flow cytometry analyses were performed at day 42 after thymic engraftment in NSG-MG mice treated with saline solution (NaCl) or anti-IL-23p19 antibody. Each point represents the mean value per experiment for each thymic biopsy obtained from one donor. Each point is from at least 4 mice. All data are from at least 4 different experiments done with thymic biopsies obtained from different AChR+ MG patients. P values were obtained with Wilcoxon matched paired test. Figure S3. Anti-IL-23p19 treatment does not induce global physiological changes in the NSG-MG mouse model. mRNA expression levels of Keratin 14 (A) in AChR+ MG thymuses engrafted in NSG-MG mice. Representative images of vascularized human MG thymuses after engraftment in mice without (B) or with treatment (C). Flow cytometry analyses of human T cells in engrafted human MG thymuses (D), in the blood (E) and in spleens (F) of NSG mice. ELISA quantification of total human immunoglobulins in the serum of NSG-MG mice (G). Images were acquired with a Zeiss Axio Observer Z1 inverted microscope using 20× magnification. In the flow cytometry graphs, each point represents the mean of the percentage of cells for an experiment. All analyses were performed at day 42 after engraftment in NSG-MG mice treated with saline solution (NaCl) or anti-IL-23p19 antibody. The data are from at least 3 different experiments performed with at least 3 thymic biopsies obtained from different MG patients. For each thymic biopsy, there were n &gt; 3 mice per treatment condition. P values were obtained using the Wilcoxon matched paired test (A–F) and an ANOVA test (G). Figure S4. Anti-IL-23p19 tends to reduce markers of eGCs in the spleens of EAMG mice. mRNA expression of Il-6 (A), Podoplanin (B) and Aid (C) in the spleens of CFA and EAMG mice treated with or without anti-IL-23p19 antibody. mRNA analyses were performed in duplicate after 2 weeks or 4 weeks of treatment by quantitative RT-PCR. For each treatment time, data were obtained from 2 independent experiments. n &gt; 4 mice per group. Each point represents an individual mouse. The mRNA results are expressed as arbitrary unit (AU) and normalized to Gapdh. P values were obtained with an ANOVA test. P value are as follows * &lt; 0.05;**0.004; ****0.0001. Figure S5. Anti-IL-23p19 modifies markers of pathogenic Th17 cells in the spleens of EAMG mice. mRNA expression of Tgf-β1 (A), Tgf-β3 (B), Il-17a (C), Il-22 (D), Il-10 (E) in the spleens of CFA and EAMG mice treated with or without anti-IL-23p19 antibody. mRNA analyses were performed in duplicate after 2 weeks or 4 weeks of treatment by quantitative RT-PCR. For each treatment time, data were obtained from 2 independent experiments. n &gt; 4 mice per group. Each point represents an individual mouse. The mRNA results are expressed as arbitrary unit (AU) and normalized to Gapdh. P values were obtained with an ANOVA test (δ = 0.05; * &lt; 0.05;**0.007; ***0.001; ****0.0001).