Journal: Immunology

Article Title: Therapeutic activity of an interleukin-4/interleukin-13 dual antagonist on oxazolone-induced colitis in mice

doi: 10.1111/imm.12319

Figure Lengend Snippet: Schematic of murine interleukin-4 (IL-4/IL-13 dual antagonist. (a) Configuration of IL-4/IL-13 dual antagonist, from N-terminal mouse IL-13Rα2 extracellular domain (ECD) to C-terminal scFv of anti-mouse IL-4 monoclonal antibody (mAb) mu11B11. (b) Schematic indicating overall structure of the IL-4/IL-13 dual antagonist, with two binding sites for each cytokine. (c) Cell-based IC50 and SPR-derived Kd values for anti-mouse IL-4 mAb mu11B11, sIL-13Rα2-Fc, and bifunctional IL-4/IL-13 antagonist. IC50 values were derived from a signal transducer and activator of transcription 6 (STAT6) phosphorylation bioassay using BaF3 cells expressing murine IL-13Rα1 and IL-4Rα.

Article Snippet: Generation and characterization of murine IL-4/IL-13 bifunctional antagonist A bifunctional molecule with specificity for murine IL-4 and IL-13 was produced on a multi-specific protein therapeutic platform (Emergent BioSolutions, Rockville, MD), as described previously.

Techniques: Binding Assay, Derivative Assay, Phospho-proteomics, Bioassay, Expressing

Journal: Immunology

Article Title: Therapeutic activity of an interleukin-4/interleukin-13 dual antagonist on oxazolone-induced colitis in mice

doi: 10.1111/imm.12319

Figure Lengend Snippet: Activity of bifunctional interleukin-4 (IL-4)/IL-13 antagonist in oxazolone-induced colitis model. Groups of 15 female SJL/J mice were sensitized with oxazolone painted on the skin on Day −5, then challenged intrarectally with oxazolone on Day 0. Starting the day before intrarectal challenge (Day −1), mice were administered bifunctional antagonist at 8, 2, or 0·5 mg/kg intraperitoneally, every 2 days. Control animals were given oxazolone but no treatment (OXA), were given OXA and treated with mouse IgG2a control, or had no disease induction (no OXA). (a) On Days 1–7, the animals were scored for body weight. At time of kiling on Day 8, colons were excised. (b) Morphology of colons at death. Representative colons are shown from animals given OXA but no treatment (OXA), those given OXA and treated with bifunctional IL-4/IL-13 antagonist (8 mg/kg) or animals that had no disease induction (no OXA). (c) Colon weight, (d) colon length; (e) ratio of colon weight/length; and (f) disease activity index (average over the 7-day study) are shown as indicators of disease severity. P-values were determined by t-test in comparison with the mouse IgG2a-treated control group.

Article Snippet: Generation and characterization of murine IL-4/IL-13 bifunctional antagonist A bifunctional molecule with specificity for murine IL-4 and IL-13 was produced on a multi-specific protein therapeutic platform (Emergent BioSolutions, Rockville, MD), as described previously.

Techniques: Activity Assay, Control, Comparison

Journal: Immunology

Article Title: Therapeutic activity of an interleukin-4/interleukin-13 dual antagonist on oxazolone-induced colitis in mice

doi: 10.1111/imm.12319

Figure Lengend Snippet: Activity of anti-interleukin-4 (IL-4), sIL-13Rα2-Fc and bifunctional IL-4/IL13 antagonist (3 mg/kg) in oxazolone-induced colitis. Female SJL/J mice were sensitized and challenged with oxazolone as described above. Starting the day before intrarectal challenge (Day −1), mice were given the protein therapeutics at 3 mg/kg intraperitoneally, every 2 days. Control animals were given oxazolone but no treatment (OXA), were given OXA and treated with mouse IgG2a control, or had no disease induction (no OXA). The no OXA group contained six animals. All other groups contained 15 animals on day 0. (a) Body weight throughout the course of treatment. P-values were determined by two-way analysis of variance in comparison with the mouse IgG2a-treated control group. (b) Colon length at time of death. P-values were determined by t-test in comparison to the mouse IgG2a-treated control group. (c) Kaplan–Meier curves, showing % survival throughout the course of treatment. In separate panels, survival of treatment groups is plotted in relation to the OXA, no OXA, and mouse IgG2a control groups; bifunctional antagonist (left), sIL-13Rα2-Fc (middle), anti-IL-4 (right). There were no statistically significant differences in survival between mice in these groups and those given mouse IgG2a control, as determined by Mantel–Cox log rank test.

Article Snippet: Generation and characterization of murine IL-4/IL-13 bifunctional antagonist A bifunctional molecule with specificity for murine IL-4 and IL-13 was produced on a multi-specific protein therapeutic platform (Emergent BioSolutions, Rockville, MD), as described previously.

Techniques: Activity Assay, Control, Comparison

Journal: Immunology

Article Title: Therapeutic activity of an interleukin-4/interleukin-13 dual antagonist on oxazolone-induced colitis in mice

doi: 10.1111/imm.12319

Figure Lengend Snippet: Interleukin-4 (IL-4) and IL-13 neutralization capacity in sera of mice from oxazolone-induced colitis model is proportional to serum concentration of bifunctional antagonist. Female SJL mice were sensitized and challenged with oxazolone as described above. Starting the day before intrarectal challenge (Day −1), mice were administered bifunctional IL-4/IL-13 antagonist at 3 or 8 mg/kg intraperitoneally, every 2 days. On Day 8, sera were collected and titres of antagonist were measured by ELISA. BaF3 cells expressing IL-4 and IL-13 receptors were incubated with sub-optimal concentrations of murine IL-4 or IL-13, in the presence of serum dilutions, for 30 min at 37°. Cells were fixed, permeabilized and stained for expression of phosphorylated signal transducer and activator of transcription 6 (STAT6). The presence of IL-4 or IL-13 neutralization activity in the sera reduced the STAT6 phosphorylation response to exogenous murine IL-4 or IL-13. (a) The relative IL-4 or IL-13 neutralization activity of individual serum samples was evaluated as the per cent inhibition of the pSTAT6 response, corrected for serum dilution. The relative potency of IL-4 and IL-13 neutralization was proportional. (b) IL-4 or IL-13 neutralization activity, plotted as a function of serum titer of IL-4/IL-13 antagonist.

Article Snippet: Generation and characterization of murine IL-4/IL-13 bifunctional antagonist A bifunctional molecule with specificity for murine IL-4 and IL-13 was produced on a multi-specific protein therapeutic platform (Emergent BioSolutions, Rockville, MD), as described previously.

Techniques: Neutralization, Concentration Assay, Enzyme-linked Immunosorbent Assay, Expressing, Incubation, Staining, Activity Assay, Phospho-proteomics, Inhibition

Journal: Immunology

Article Title: Therapeutic activity of an interleukin-4/interleukin-13 dual antagonist on oxazolone-induced colitis in mice

doi: 10.1111/imm.12319

Figure Lengend Snippet: Serum concentrations of serum amyloid P (SAP) are associated with disease activity in the oxazolone-induced colitis model. Female SJL mice were sensitized and challenged with oxazolone. Starting the day before intrarectal challenge (Day −1), mice were administered bifunctional interleukin-4 (IL-4) /IL-13 antagonist, at 8, 3, or 0·5 mpk every 2 days. At time of killing on Day 7, sera were collected and assayed for SAP by ELISA. Associations between these readouts and disease correlates are shown for all animals in the study, across all treatment groups: (a) average body weight versus serum SAP; (b) colon length versus serum SAP; (c) serum bifunctional IL-4/IL-13 antagonist concentration versus colon length; (d) serum concentrations of bifunctional IL-4/IL-13 antagonist versus SAP for the 0·5 mpk dose group. R2 and P-values were derived from linear regression analysis.

Article Snippet: Generation and characterization of murine IL-4/IL-13 bifunctional antagonist A bifunctional molecule with specificity for murine IL-4 and IL-13 was produced on a multi-specific protein therapeutic platform (Emergent BioSolutions, Rockville, MD), as described previously.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Concentration Assay, Derivative Assay

Journal: Immunology

Article Title: Therapeutic activity of an interleukin-4/interleukin-13 dual antagonist on oxazolone-induced colitis in mice

doi: 10.1111/imm.12319

Figure Lengend Snippet: Colon gene expression changes are proportional to serum concentration of bifunctional interleukin-4 (IL-4) /IL-13 antagonist. Gene expression data were plotted against the concentration of bifunctional antagonist in the serum for individual animals given a 0·5 mg/kg dose. R2 and P-values are shown as measures of correlation and significance.

Article Snippet: Generation and characterization of murine IL-4/IL-13 bifunctional antagonist A bifunctional molecule with specificity for murine IL-4 and IL-13 was produced on a multi-specific protein therapeutic platform (Emergent BioSolutions, Rockville, MD), as described previously.

Techniques: Gene Expression, Concentration Assay