migg1 isotype control ab  (Eli Lilly)

Journal: The Journal of Clinical Investigation

Article Title: Angiopoietin-2 blockade ameliorates autoimmune neuroinflammation by inhibiting leukocyte recruitment into the CNS

doi: 10.1172/JCI130308

Figure Lengend Snippet: ( A ) Ang2 protein concentration in the serum and SC lysates at different time points after EAE induction (0 dpi: n = 4; 7 dpi: n = 4; 14 dpi: n = 5; 21 dpi: n = 4; 28 dpi: n = 3). ( B ) Clinical scores and percentage of body weight loss of control (Ctrl, n = 9) versus EC-Ang2 ( n = 11) mice induced with active EAE. ( C ) Clinical scores and percentages of body weight loss of mice induced with active EAE and treated with mIgG1 versus Ang2 Ab prophylactically (starting at the time of EAE induction; 0 dpi) ( n = 10 per group). ( D ) Clinical scores of mice induced with active EAE and treated with mIgG1 versus Ang2 Ab preemptively (starting during the effector phase of EAE at 7 dpi) ( n = 10 per group). ( E and F ) Representative images and quantifications of MBP staining to show loss of myelin in the SC white matter from both prophylactic 14 dpi and preemptive 28 dpi groups ( n = 10 per group). Scale bars: 100 μm. ( G ) Clinical scores of mice induced with adoptive transfer EAE and treated with mIgG1 versus Ang2 Ab starting at the time of adoptive transfer. Data are pooled from 2 independent experiments ( n = 16 per group). Arrows indicate Ab injections. Mean ± SEM, 1-way ANOVA with Dunnett’s post hoc test for multiple comparisons ( A ), nonparametric Mann-Whitney U test ( B - D , and G , comparison of AUC values of clinical EAE scores over the disease course), 2-way repeated measures ANOVA ( B and C , body weight loss), and 2-tailed Student’s t test ( E and F ). * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: mIgG1 isotype control Ab (Eli Lilly and Co.), mouse anti-mouse Ang2 Ab (18E5, Eli Lilly and Co.), hIgG1 isotype control Ab (Synagis, AbbVie), and ABTAA ( ) were administrated i.p. at a dose of 20–25 mg/kg body weight 2 to 3 times a week.

Techniques: Protein Concentration, Control, Staining, Adoptive Transfer Assay, MANN-WHITNEY, Comparison

Journal: The Journal of Clinical Investigation

Article Title: Angiopoietin-2 blockade ameliorates autoimmune neuroinflammation by inhibiting leukocyte recruitment into the CNS

doi: 10.1172/JCI130308

Figure Lengend Snippet: ( A ) Flow cytometric quantification of the number of immune cells in the SCs of mIgG1- versus Ang2 Ab–treated EAE mice at 12 dpi ( n = 10 per group). ( B ) RT-qPCR quantification of mRNA levels of Th signature cytokines ( Ifng , Tnf , Il4 , and Il17a ) and integrin subunits ( Itga4 and Itgb1 ) in the SCs of mIgG1- versus Ang2 Ab–treated EAE mice at 14 dpi ( n = 10 per group). ( C ) Flow cytometric quantification of the number of immune cells in the SCs of control ( n = 6) versus EC-Ang2 ( n = 9) EAE mice at 12 dpi. ( D and E ) Representative immunofluorescent images and quantifications of Iba1 + microglia and macrophages, Ly-6G + granulocytes, and CD4 + Th cells in the SCs of mIgG1- versus Ang2 Ab–treated control and EAE mice ( n = 10 per group) at 14 dpi as well as in the SCs of control ( n = 7) versus EC-Ang2 ( n = 8) control and EAE mice at 12 dpi. Scale bars: 100 μm. Mean ± SEM, 2-tailed Student’s t test ( A - E ), and 2-way ANOVA with Bonferroni’s post hoc test for multiple comparisons (Iba1 staining in D and E ). * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: mIgG1 isotype control Ab (Eli Lilly and Co.), mouse anti-mouse Ang2 Ab (18E5, Eli Lilly and Co.), hIgG1 isotype control Ab (Synagis, AbbVie), and ABTAA ( ) were administrated i.p. at a dose of 20–25 mg/kg body weight 2 to 3 times a week.

Techniques: Quantitative RT-PCR, Control, Staining

Journal: The Journal of Clinical Investigation

Article Title: Angiopoietin-2 blockade ameliorates autoimmune neuroinflammation by inhibiting leukocyte recruitment into the CNS

doi: 10.1172/JCI130308

Figure Lengend Snippet: ( A ) T-distributed stochastic neighbor embedding (t-SNE) analysis of main immune cell clusters in the SCs of mIgG1- versus Ang2 Ab– treated EAE mice at 14 dpi and single transgenic control versus EC-Ang2 EAE mice at 12 dpi. Heatmap showing log 2 expression of known marker genes for each cluster. ( B and C ) Relevant GO biological processes of significantly (adjusted P < 0.05) downregulated genes after Ang2 blockade and upregulated genes after Ang2 overexpression in microglia ( B ) and macrophages ( C ). ( D and E ) Venn diagram illustrating the number of genes that were regulated by Ang2 in microglia and macrophages. ( F ) Violin plots showing mRNA expression of significantly (adjusted P < 0.05) downregulated APOE-induced and MHCII-associated genes after Ang2 blockade. IgG, mIgG1; A2, Ang2 Ab. ( G ) Representative images and quantifications of MHCII immunostaining in Iba1 + cells in the SCs of mIgG1- versus Ang2 Ab–treated EAE mice at 14 dpi ( n = 10 per group) and control ( n = 7) versus EC-Ang2 ( n = 8) EAE mice at 12 dpi. Scale bars: 100 μm. ( H ) Representative flow cytometry overlay plots and quantifications showing GMFI of MHCII expression in microglia and macrophages in the SCs of mIgG1- versus Ang2 Ab–treated EAE mice at 14 dpi ( n = 10 per group) and control versus EC-Ang2 EAE mice at 12 dpi ( n = 8 per group). Mean ± SEM, 2-tailed Student’s t test ( G and H ). * P < 0.05; ** P < 0.01.

Article Snippet: mIgG1 isotype control Ab (Eli Lilly and Co.), mouse anti-mouse Ang2 Ab (18E5, Eli Lilly and Co.), hIgG1 isotype control Ab (Synagis, AbbVie), and ABTAA ( ) were administrated i.p. at a dose of 20–25 mg/kg body weight 2 to 3 times a week.

Techniques: Transgenic Assay, Control, Expressing, Marker, Over Expression, Immunostaining, Flow Cytometry

Journal: The Journal of Clinical Investigation

Article Title: Angiopoietin-2 blockade ameliorates autoimmune neuroinflammation by inhibiting leukocyte recruitment into the CNS

doi: 10.1172/JCI130308

Figure Lengend Snippet: ( A ) Violin plots showing expression of Angpt2 and its receptors Tek , Itga5 , and Itgb1 in EC, microglia, and macrophage clusters of mIgG1- and Ang2 Ab–treated EAE mice. ( B ) GMFI of Tie2 and α 5 integrin on the surface of ECs, macrophages, and microglia from naive versus EAE mice (naive, n = 4; EAE, n = 3) at 14 dpi as analyzed by flow cytometry. ( C ) Flow cytometric analysis of active cell surface integrin (FN7-10 binding) relative to total cell surface α 5 β 1 integrin in microglia and macrophages from mIgG1- and Ang2 Ab–treated EAE mice (mIgG1, n = 8; Ang2 Ab, n = 7) at the disease peak (16 dpi). Mean ± SEM, 2-way ANOVA with Bonferroni’s post hoc test for multiple comparisons ( B ) and 2-tailed Student’s t test ( C ). * P < 0.05; *** P < 0.001.

Article Snippet: mIgG1 isotype control Ab (Eli Lilly and Co.), mouse anti-mouse Ang2 Ab (18E5, Eli Lilly and Co.), hIgG1 isotype control Ab (Synagis, AbbVie), and ABTAA ( ) were administrated i.p. at a dose of 20–25 mg/kg body weight 2 to 3 times a week.

Techniques: Expressing, Flow Cytometry, Binding Assay

Journal: The Journal of Clinical Investigation

Article Title: Angiopoietin-2 blockade ameliorates autoimmune neuroinflammation by inhibiting leukocyte recruitment into the CNS

doi: 10.1172/JCI130308

Figure Lengend Snippet: ( A and B ) t-SNE analysis of the main EC clusters (venous and arterial) in the SCs of mIgG1- versus Ang2 Ab–treated EAE mice at 14 dpi and single transgenic control versus EC-Ang2 EAE mice at 12 dpi. capillary-V, capillary-venous; capillary-A, capillary-arterial. ( C ) Heatmap showing log 2 expression of known marker genes in each cluster. ( D and E ) Relevant GO biological processes of significantly (adjusted P < 0.05) downregulated genes after Ang2 blockade ( D ) and upregulated genes after Ang2 overexpression ( E ) in the SC capillary-venous ECs. ( F ) Venn diagram illustrating the number of genes that were regulated by Ang2 in the SC capillary-venous ECs. ( G ) Violin plots showing Vcam1 expression in different EC clusters as well as its differential expression in capillary-venous ECs regulated by Ang2. ( H and I ) Representative images (EAE) and quantification of VCAM1 in the SC blood vessels of mIgG1- versus Ang2 Ab–treated control ( n = 6 per group) and EAE ( n = 10 per group) mice at 14 dpi and control versus EC-Ang2 control (Ctrl, n = 3; EC-Ang2 , n = 4) and EAE (Ctrl, n = 7; EC-Ang2 , n = 8) mice at 12 dpi. Scale bars: 100 μm. Mean ± SEM, 2-way ANOVA with Bonferroni’s post hoc test for multiple comparisons ( H and I ). *** P < 0.001.

Article Snippet: mIgG1 isotype control Ab (Eli Lilly and Co.), mouse anti-mouse Ang2 Ab (18E5, Eli Lilly and Co.), hIgG1 isotype control Ab (Synagis, AbbVie), and ABTAA ( ) were administrated i.p. at a dose of 20–25 mg/kg body weight 2 to 3 times a week.

Techniques: Transgenic Assay, Control, Expressing, Marker, Over Expression, Quantitative Proteomics

Journal: The Journal of Clinical Investigation

Article Title: Angiopoietin-2 blockade ameliorates autoimmune neuroinflammation by inhibiting leukocyte recruitment into the CNS

doi: 10.1172/JCI130308

Figure Lengend Snippet: ( A and B ) Representative images and quantifications of Evans blue leakage in the SCs of naive ( n = 3), mIgG1- versus Ang2 Ab–treated EAE mice ( n = 5 per group) and control versus EC-Ang2 control ( n = 6 per group) and EAE ( n = 7 per group) mice at 12 dpi. ( C ) Representative images and quantification of extravascular TER-119 + RBCs in the SCs of control ( n = 6) versus EC-Ang2 ( n = 8) EAE mice at 12 dpi. Scale bars: 100 μm. Mean ± SEM, 1- or 2-way ANOVA with Bonferroni’s post hoc test for multiple comparisons ( A and B ) and 2-tailed Student’s t test ( C ). * P < 0.05; ** P < 0.01; *** P < 0.001.

Article Snippet: mIgG1 isotype control Ab (Eli Lilly and Co.), mouse anti-mouse Ang2 Ab (18E5, Eli Lilly and Co.), hIgG1 isotype control Ab (Synagis, AbbVie), and ABTAA ( ) were administrated i.p. at a dose of 20–25 mg/kg body weight 2 to 3 times a week.

Techniques: Control