anti human il 12p40  (Bio X Cell)

Journal: Science Advances

Article Title: Inflammatory arthritis irAE may represent a unique autoimmune disease primarily driven by T cells but likely not autoantibodies

doi: 10.1126/sciadv.aea4262

Figure Lengend Snippet: ( A to G ) PBMCs from the patients with irAE were cultured in the plate coated with anti-CD3 and anti-CD28 (10 μg/ml) in the presence of IgG1 isotype control or anti–human IL-6R (50 μg/ml), anti–human IL-12p40 (50 μg/ml), and anti–human IFNAR1 (50 μg/ml) for 3 days; n = 9. (A) Expression of CD38 and CD127 on CD8 + T cells. Right: Percentage of CD38 + CD127 − CD8 + T cells. (B and C) CD8 + T cells activated for 5 days were restimulated with PMA, ionomycin, and monensin for 5 hours. (B) Expression of granzyme B and IFN-γ. Right: Percentage of granzyme B + IFN-γ + CD8 + T cells. (C) Expression of perforin and IFN-γ. Right: Percentage of perforin + IFN-γ + CD8 + T cells. [(D) and (E)] MFIs of GluCy5 (D), TMRM, and MTDR (E) in CD8 + T cells were presented. [(F) and (G)] CD4 + T cells activated for 5 days were restimulated with PMA, ionomycin, and monensin for 5 hours. (F) Expression of IL-21 and IL-2. Right: Percentage of IL-21 + IL-2 + CD4 + T cells. (G) Expression of CD38 and CXCR5 on CD4 + T cells. Right: Percentage of CD38 + CXCR5 − CD4 + T cells. Data in graphs represent mean ± SEM. Significance was tested paired Student’s t test [(A) to (G)].

Article Snippet: PBMCs from patients with irAE were thawed and rested in a complete medium for 1 hour and then activated with plate-coated anti–human CD3 and anti–human CD28 (10 μg/ml) with IgG1 isotype control (Bio X Cell, catalog no. CP174) or a combination of anti–human IL-6R 50 μg/ml; Bio X Cell, catalog no. SIM0014), anti–human IL-12p40 (50 μg/ml; Bio X Cell, catalog no. SIM0020), and anti–human IFNAR1 (50 μg/ml; Bio X Cell, catalog no. SIM0022) for 3 days.

Techniques: Cell Culture, Control, Expressing

Journal: Immunity

Article Title: Structural activation of pro-inflammatory human cytokine IL-23 by cognate IL-23 receptor enables recruitment of the shared receptor IL-12Rβ1

doi: 10.1016/j.immuni.2017.12.008

Figure Lengend Snippet: A) Schematic representation of protein components participating in IL-23 and IL-12 signaling complexes. EC: Extracellular, PM: Plasma Membrane, CP: Cytoplasm, p19: IL-23p19, p40: IL-12p40. B) Cartoon representation of the IL-23:IL-23R crystal structure. Nanobody 22E11 used as a crystallization adjuvant is shown in gray surface representation. C) Close-up view of the IL-23R:IL-23p19 interface in the vicinity of W156 in IL-23p19. D) Restructuring of IL-23 upon binding to IL-23R. E) Close-up view of the interactions around the AB-loop of IL-23p19. F) Top-down view of the IL-23:IL-23R complex with IL-23R (surface). See also Figure S1, Figure S2, Figure S3, Table S1, and Table S2.

Article Snippet: The presence of both IL-12p40 and IL-12Rβ1 in the crosslinked species was ascertained by western blot using polyclonal anti human IL-12p40 and anti-human IL-12 Rβ1 primary antibodies (R&D).

Techniques: Crystallization Assay, Binding Assay

Journal: Frontiers in Immunology

Article Title: Expression and Function of IL12/23 Related Cytokine Subunits (p35, p40, and p19) in Giant-Cell Arteritis Lesions: Contribution of p40 to Th1- and Th17-Mediated Inflammatory Pathways

doi: 10.3389/fimmu.2018.00809

Figure Lengend Snippet: IL-12/23p40, IL-12p35, and IL-23p19 concentrations in temporal artery biopsies (mRNA) and in serum from patients with giant-cell arteritis (GCA). (A) IL-12/23p40 mRNA, (B) IL-23p19 mRNA, and (C) IL-12p35 mRNA expression (relative units) in temporal artery biopsies from 36 treatment-naïve patients and 20 controls. (D) IL-12/23p40 concentration in sera from 36 treatment-naïve patients and 19 controls.

Article Snippet: Temporal artery sections from 10 GCA patients and 10 controls were embedded in Matrigel to ensure prolonged survival and cultured ex vivo as described ( , , , ) with or without neutralizing anti-human IL-12p40 mouse monoclonal Ab (10 μg/ml R&D Systems), or dexamethasone (DXM) (0.5 μg/ml, Sigma-Aldrich).

Techniques: Expressing, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Expression and Function of IL12/23 Related Cytokine Subunits (p35, p40, and p19) in Giant-Cell Arteritis Lesions: Contribution of p40 to Th1- and Th17-Mediated Inflammatory Pathways

doi: 10.3389/fimmu.2018.00809

Figure Lengend Snippet: Immunofluorescence detection of IL-12/23p40, IL-12p35, and IL-23p19 subunit expression in temporal artery lesions. (A) IL-12p35 staining (blue) in a temporal artery from a control individual showing almost selective expression in the media layer. Nuclei were stained with Hoechst (white). (B) IL-12p35 expression in GCA-involved temporal artery section, predominantly in inflammatory infiltrates. (C,D) Negative IL-23p19 immunostaining (green) in sections of normal temporal arteries (C) and intense expression in GCA samples (D) where IL-23p19 expression can be observed in all arterial layers especially in the most inflamed areas. (E) Lack of IL-12/23p40 immunostaining in a temporal artery from a control. (F) Detection of IL-12/23p40 expression (red) in a GCA-involved temporal artery section predominantly in the adventitial layer. (G,H) IL-12p35 (blue), IL-23p19 (green) and IL-12/23p40 (red) staining merge in a temporal artery section from a control and from a GCA patient, respectively. Pictures are representative of four arteries from four GCA patients and two controls, and at least three sections per sample were evaluated.

Article Snippet: Temporal artery sections from 10 GCA patients and 10 controls were embedded in Matrigel to ensure prolonged survival and cultured ex vivo as described ( , , , ) with or without neutralizing anti-human IL-12p40 mouse monoclonal Ab (10 μg/ml R&D Systems), or dexamethasone (DXM) (0.5 μg/ml, Sigma-Aldrich).

Techniques: Immunofluorescence, Expressing, Staining, Control, Immunostaining

Journal: Frontiers in Immunology

Article Title: Expression and Function of IL12/23 Related Cytokine Subunits (p35, p40, and p19) in Giant-Cell Arteritis Lesions: Contribution of p40 to Th1- and Th17-Mediated Inflammatory Pathways

doi: 10.3389/fimmu.2018.00809

Figure Lengend Snippet: Coexpression of subunits to conform IL-12 and IL-23 in temporal artery lesions from patients with GCA. (A) Close-up view of a GCA-involved artery where IL-12p35 + cells (blue) can be observed predominantly in media layer, IL-23p19 + cells (green) in all arterial layers and IL-12/23p40 + cells (red) in the adventitia. (B) Higher magnification of inflammatory cells expressing IL-12/23p40 (red), (C) IL-23p19 (green), and (D) merge showing expression of IL-23 (orange, indicated by arrows). (E) Higher magnification of inflammatory cells expressing IL-12/23p40 (red), (F) IL12p35 (blue), and (G) merge showing expression of IL-12 (purple, indicated by arrows). The temporal arteries were subjected to proximity ligation assay (PLA) to determine proximity of IL-12 and IL-23 subunits. The red staining marks the colocalization (<40 nm) of IL-12/23p40 and IL-12p35 (IL-12). Blue (Hoechst) marks the cell nuclei. (H) Negative PLA signalfor colocalization of IL-12/23p40 and IL-12p35 (IL-12) in a control artery. Minimal autofluorescence of the elastic lamina can be appreciated (I) . Positive colocalization of IL-12/23p40 and IL-12p35 (IL-12) in a GCA-affected temporal artery. The boxed area is magnified on the right (J) . (K) Negative PLA signal for colocalization of IL-12/23p40 and IL-23p19 (IL-23) in a control artery. Minimal autofluorescence of the elastic lamina can be appreciated (L) Colocalization of IL-12/23p40 and IL-23p19 (IL-23) in a positive GCA temporal artery. The boxed area is magnified on the right (M) .

Article Snippet: Temporal artery sections from 10 GCA patients and 10 controls were embedded in Matrigel to ensure prolonged survival and cultured ex vivo as described ( , , , ) with or without neutralizing anti-human IL-12p40 mouse monoclonal Ab (10 μg/ml R&D Systems), or dexamethasone (DXM) (0.5 μg/ml, Sigma-Aldrich).

Techniques: Expressing, Proximity Ligation Assay, Staining, Control

Journal: Frontiers in Immunology

Article Title: Expression and Function of IL12/23 Related Cytokine Subunits (p35, p40, and p19) in Giant-Cell Arteritis Lesions: Contribution of p40 to Th1- and Th17-Mediated Inflammatory Pathways

doi: 10.3389/fimmu.2018.00809

Figure Lengend Snippet: Decreased IL-12/23p40 and IL-23p19 expression in temporal arteries from treated GCA patients. IL-23p19 (A) , IL-12/23p40 (B) , and IL-12p35 (C) expression (relative units) in temporal arteries from 36 treatment-naïve and 14 prednisone-treated GCA patients. (D) mRNA IL-12/23p40 expression and (E) mRNA IL-23p19 expression on cultured biopsies (negative and GCA) , showing the effect of dexamethasone (DXM) in GCA arteries. Bars represent mean ± SEM.

Article Snippet: Temporal artery sections from 10 GCA patients and 10 controls were embedded in Matrigel to ensure prolonged survival and cultured ex vivo as described ( , , , ) with or without neutralizing anti-human IL-12p40 mouse monoclonal Ab (10 μg/ml R&D Systems), or dexamethasone (DXM) (0.5 μg/ml, Sigma-Aldrich).

Techniques: Expressing, Cell Culture

Journal: Frontiers in Immunology

Article Title: Expression and Function of IL12/23 Related Cytokine Subunits (p35, p40, and p19) in Giant-Cell Arteritis Lesions: Contribution of p40 to Th1- and Th17-Mediated Inflammatory Pathways

doi: 10.3389/fimmu.2018.00809

Figure Lengend Snippet: IL-12/23p40, IL-12/23p19, and IL-12p35 mRNA expression in lesions and long-term response to therapy. Percentage of treatment-naïve GCA patients requiring prednisone treatment over time, according to the intensity of IL-12/23p40 (A) , IL-12/23p19 (B) , and IL-12p35 (C) mRNA expression, in temporal artery lesions. High refers to mRNA levels above the 75th percentile ( N = 26), and low below the 75th percentile ( N = 10). IL-12/23p40 (D) , IL-23p19 (E) , and IL-12p35 (F) mRNA concentrations in initial temporal artery biopsies from patients still requiring prednisone ( N = 12) compared with patients in sustained remission ( N = 24), 3 years after diagnosis.

Article Snippet: Temporal artery sections from 10 GCA patients and 10 controls were embedded in Matrigel to ensure prolonged survival and cultured ex vivo as described ( , , , ) with or without neutralizing anti-human IL-12p40 mouse monoclonal Ab (10 μg/ml R&D Systems), or dexamethasone (DXM) (0.5 μg/ml, Sigma-Aldrich).

Techniques: Expressing, Biomarker Discovery

Journal: Frontiers in Immunology

Article Title: Expression and Function of IL12/23 Related Cytokine Subunits (p35, p40, and p19) in Giant-Cell Arteritis Lesions: Contribution of p40 to Th1- and Th17-Mediated Inflammatory Pathways

doi: 10.3389/fimmu.2018.00809

Figure Lengend Snippet: Changes in gene expression by blocking IL-12/23p40 on cultured GCA biopsies. mRNA concentrations of IFNγ, CXCL11, CXCL10, CXCL9, IL-17, IL-6, IL-1β, and tumor necrosis factor α in 10 cultured control arteries (negative biopsies, Neg Bx) vs 10 cultured GCA-involved arteries untreated or exposed to anti-human IL-12p40 mouse monoclonal Ab (anti-p40) (10 µg/ml), or dexamethasone (DXM) (0.5 µg/ml). Statistical comparisons were performed between histologically negative and GCA-involved arteries and between GCA-involved arteries and anti-p40 treated and DXM treated. Bars represent mean ± SEM, * p < 0.05.

Article Snippet: Temporal artery sections from 10 GCA patients and 10 controls were embedded in Matrigel to ensure prolonged survival and cultured ex vivo as described ( , , , ) with or without neutralizing anti-human IL-12p40 mouse monoclonal Ab (10 μg/ml R&D Systems), or dexamethasone (DXM) (0.5 μg/ml, Sigma-Aldrich).

Techniques: Gene Expression, Blocking Assay, Cell Culture, Control