il13ra2  (R&D Systems)

Journal: Cell reports

Article Title: TGF-β1-mediated intercellular signaling fuels cooperative cellular invasion

doi: 10.1016/j.celrep.2025.115315

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Article Snippet: CD213a2 (IL13Ra2) (APC) , Miltenyi Biotec , Cat#130-128-220; RRID: AB_2928320.

Techniques: Recombinant, Reverse Transcription, Bicinchoninic Acid Protein Assay, Staining, RNA Sequencing

Journal: bioRxiv

Article Title: A Bi-Specific T Cell-Engaging Antibody Shows Potent Activity, Specificity, and Tumor Microenvironment Remodeling in Experimental Syngeneic and Genetically Engineered Models of GBM

doi: 10.1101/2024.12.18.628714

Figure Lengend Snippet: ( A ) Schematic representation of BTE. BTE consisted of the scFv targeting murine CD3E specific, derived from 2C11-145, and the scFv targeting IL13RA2, derived from clone47, fused together via a flexible GlyS (Gly4Ser) linker. Negative control BTE (NC-BTE) differed from BTE in the VH domain of IL13RA2-scFv, which was swapped with the VH domain of scFv p588 of the non-specific MOPC21 antibody. ( B ) A western blot of affinity-purified BTE was detected using anti-His HRP conjugated antibodies, and a specific single band ∼was observed at 55 kDa. ( C ) ELISA assay of affinity purified BTE and NC-BTE demonstrating dose-dependent and specific binding of BTE, but not NC-BTE, to recombinant IL13RA2, but not to IL13RA1 ( D ) Chromium-51 (Cr51) release assay of murine T cells and murine glioma cells treated with BTE (red), NC-BTE (white), or Dynabeads™ Mouse T-Activator CD3/CD28 (grey) showing BTE, but not NC-BTE, induced dose-dependent killing of IL13Rα2-expressing SMA560 and GL261 murine glioma cells (Effector-to-target [E : T] ratio = 20:1; Incubation time = 24 hr; n=3/group). ( E ) Flow cytometric analysis of murine T cells co-cultured with IL13Rα2-expressing SMA560, GL261, and CT2A murine glioma cells showing BTE, but not NC-BTE induced robust expression T cell activation markers CD69 and CD25 (Effector-to-target [E: T] ratio = 20:1; Incubation time = 24 hr; n=3/group). One-way ANOVA followed by Tukey’s Honest Significant Difference (HSD) test for pairwise comparisons)

Article Snippet: The following antibodies were used: IL13RA2 (R&D Systems, Cat No. AF146), Olig2 (Abcam, ab109186), Ki67 (Abcam, ab16667), CD3 (Abcam, ab16669), and CD11B (Abcam, ab133357).

Techniques: Derivative Assay, Negative Control, Western Blot, Affinity Purification, Enzyme-linked Immunosorbent Assay, Binding Assay, Recombinant, Release Assay, Expressing, Incubation, Cell Culture, Activation Assay

Journal: bioRxiv

Article Title: A Bi-Specific T Cell-Engaging Antibody Shows Potent Activity, Specificity, and Tumor Microenvironment Remodeling in Experimental Syngeneic and Genetically Engineered Models of GBM

doi: 10.1101/2024.12.18.628714

Figure Lengend Snippet: ( A ) Dynamic live fluorescent imaging of AF647-labeled BTE showing penetration and persistence in the brains of GL261-IL13RA2 tumor-bearing mice (Fluorescent signal measured as radiance [p/s/cm2/sr]; Image acquisition time = 3, 6, 24, 48, and 72 hrs post AF647-BTE injection; n=3). ( B ) Kaplan–Meier survival curve of SMA.560-IL13RA2 tumor-bearing mice treated with either BTE (200 µg/animal) (red), NC-BTE (200 µg/animal) (blue), or saline (black). Wald test from Cox proportional hazards regression analysis ( C ) Experimental setup used in flow cytometric analysis of the brains of SMA.560-IL13RA2 tumor-bearing mice following treatment with either BTE or NC-BTE (n=5/group). Flow cytometric analysis revealed ( D ) no change in CD4 helper t cell (CD45+ CD4+), ( E ) increase in regulatory T cell (CD45+ CD4+ FOXP3+), ( F ) increase in CD8 T cell (CD45+ CD8+), ( G ) increase in memory-like CD8 T cell (CD45+CD8+CD127HiKLRG1−), ( H ) no change in short-lived memory CD8 T cells (CD45+ CD8+ CD127Hi KLRG1+), ( I ) increase in tissue-resident memory CD8 T cells (CD45+ CD8+ CD127Hi KLRG1− CD103+), ( J ) increase in monocytic MDSC (mo.MDSC) (CD45+ CD11B+ CD11C−LY6G− LY6C+), ( K ) no change in polymorphonuclear MDSC (PMN.MDSC) (CD45+ CD11B+ CD11C− LY6G+ LY6C+), ( L ) no change in dendritic cells (CD45+ CD11B+ CD11C+). One-way ANOVA followed by Tukey’s Honest Significant Difference (HSD) test for pairwise comparisons).

Article Snippet: The following antibodies were used: IL13RA2 (R&D Systems, Cat No. AF146), Olig2 (Abcam, ab109186), Ki67 (Abcam, ab16667), CD3 (Abcam, ab16669), and CD11B (Abcam, ab133357).

Techniques: Imaging, Labeling, Injection, Saline

Journal: bioRxiv

Article Title: A Bi-Specific T Cell-Engaging Antibody Shows Potent Activity, Specificity, and Tumor Microenvironment Remodeling in Experimental Syngeneic and Genetically Engineered Models of GBM

doi: 10.1101/2024.12.18.628714

Figure Lengend Snippet: ( A ) Histopathological analysis of RCAS-P53fl/fl-PTENfl/fl-IL13RA2 GEMM glioma model with high magnification images showing morphological characteristics similar to human GBM. H&E (top-left) shows areas of pseudopalisading necrosis (i), microvascular proliferation (ii), and hyperplasia (iii). IHC staining revealed many infiltrating CD11B+ macrophages (top-right), scarce infiltrating CD3+ T cells (bottom-right), and heterogeneous IL13RA2 expression (bottom-left) within the tumor bed. ( B ) Kaplan–Meier survival curve of de novo GEMM glioma model treated with BTE (red) (n=20) or saline (black) (n=19). Animals were followed for survival, and statistical significance was tested via Wald test from Cox proportional hazards regression analysis. ( C ) An experimental setup was used in flow cytometric analysis of the brains and spleens of the GEMM glioma model following treatment with either BTE or saline (n=5/group). Flow cytometric analysis revealed ( D ) no change in regulatory T cell (CD45+ CD4+ FOXP3+) and brain-specific increases in ( E ) activated CD8 T cell (CD45+ CD8+ CD69+), ( F ) cytotoxic CD8 T cell (CD45+ CD8+ CD69+), ( H ) resident CD8 T cells (CD45+ CD8+ CD44+ CD62L-), and ( G ) tissue-resident memory CD8 T cells (CD45+ CD8+ CD44+ CD62L- CD103+).

Article Snippet: The following antibodies were used: IL13RA2 (R&D Systems, Cat No. AF146), Olig2 (Abcam, ab109186), Ki67 (Abcam, ab16667), CD3 (Abcam, ab16669), and CD11B (Abcam, ab133357).

Techniques: Immunohistochemistry, Expressing, Saline

Journal: bioRxiv

Article Title: IL-13/IL-13Rα2 axis promotes proliferation of angiosarcoma cells

doi: 10.1101/2024.10.24.619789

Figure Lengend Snippet: (A) Relative mRNA expression of IL13RA1 (A) and IL13RA2 (B) in various cell types. (C) Surface protein expression levels of IL-13Ra2 in MO-LAS-B and A431 cells. Representative images from immunohistochemical staining for IL-13Ra2 (D, E) and IL-13 (F-H) in angiosarcoma (D, F), hemangioma (E), atopic dermatitis (G), and healthy skin (H) samples. A subset of atypical tumor cells forming luminal structures (red arrows). Scale bars = 50μm.

Article Snippet: MO-LAS-B cells were grown to 40% confluence in 10cm-dishes before transfection with Silencer Select Human IL13RA2 siRNA (s7375, Thermo Fisher Scientific, Waltham, MA, USA) or Silencer Select Negative Control No.1 siRNA (4390843, Thermo Fisher Scientific) using Lipofectamine 3000 Transfection Reagent (L3000001, Thermo Fisher Scientific).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: bioRxiv

Article Title: IL-13/IL-13Rα2 axis promotes proliferation of angiosarcoma cells

doi: 10.1101/2024.10.24.619789

Figure Lengend Snippet: (A, B) The effect of IL-13 (10 ng/mL) for 24 hours on MO-LAS-B cell numbers was evaluated by flow cytometry (A) and Cell Counting Kit-8 cell proliferation assay (B) ( n = 6 for each group). (C) Percent of bromodeoxyuridine (BrdU)-positive cells after vehicle or IL-13 (10 ng/mL) treatment for 48 hours ( n = 3 for each group). (D) Representative flow cytometric analysis of BrdU-positive cells from the vehicle or IL-13 treated cells. Numbers show percentages of cells in the indicated gate. (E) Percent of Ki-67 hi cells after vehicle or IL-13 (10 ng/mL) treatment for 48 hours ( n = 6 for each group). (F) Representative histograms from Ki-67 staining of angiosarcoma cells, MO-LAS-B. Percent of Ki-67high cells in each sample was indicated. Two histograms in this figure were representative of 6 samples in each group. (G) Cell proliferation assay demonstrating the effects of IL-13-IL13Rα2 axis on the MO-LAS-B cell numbers. All populations were treated with IL-13 (10 ng/mL) for 72 hours. Some samples were transfected with scrambled siRNAs or IL13RA2 siRNAs 48 hours before IL-13 stimulation. Some samples were cultured with anti-IL-13 neutralizing antibodies 1 hour before IL-13 stimulation ( n = 6 for each group). (A, B, C, E, G) Representative data from one of at least two independent experiments are shown. * p <0.05, *** p <0.001, **** p <0.0001.

Article Snippet: MO-LAS-B cells were grown to 40% confluence in 10cm-dishes before transfection with Silencer Select Human IL13RA2 siRNA (s7375, Thermo Fisher Scientific, Waltham, MA, USA) or Silencer Select Negative Control No.1 siRNA (4390843, Thermo Fisher Scientific) using Lipofectamine 3000 Transfection Reagent (L3000001, Thermo Fisher Scientific).

Techniques: Flow Cytometry, Cell Counting, Proliferation Assay, Staining, Transfection, Cell Culture

Journal: bioRxiv

Article Title: IL-13/IL-13Rα2 axis promotes proliferation of angiosarcoma cells

doi: 10.1101/2024.10.24.619789

Figure Lengend Snippet: (A, C) Relative mRNA expression of IL13RA2 (A) and VEGFA (C) under vehicle or IL-13 treatment (10 ng/mL). Some samples were treated with STAT6 inhibitor, AS1517499, 1 hour before the vehicle or IL-13 stimulation, ( n = 6 for each group). (B) CCK-8 cell proliferation assay evaluating the effect of VEGF-A on MO-LAS-B cell proliferation, MO-LAS-B cells were stimulated with vehicle or VEGF-A (50ng/ml) for 48 hours ( n = 8 for each group). (D) Conceptual model for the role of IL-13 in the proliferation of angiosarcoma cells. IL-13Rα2 is highly expressed in angiosarcoma, and IL-13 promotes proliferation through IL-13Rα2. Moreover, IL-13Rα2 and VEGF expression are upregulated by IL-13 stimulation in a STAT6-dependent manner. (A-C) Representative data from one of at least two independent experiments are shown. ** p <0.01, *** p <0.001, **** p <0.0001.

Article Snippet: MO-LAS-B cells were grown to 40% confluence in 10cm-dishes before transfection with Silencer Select Human IL13RA2 siRNA (s7375, Thermo Fisher Scientific, Waltham, MA, USA) or Silencer Select Negative Control No.1 siRNA (4390843, Thermo Fisher Scientific) using Lipofectamine 3000 Transfection Reagent (L3000001, Thermo Fisher Scientific).

Techniques: Expressing, CCK-8 Assay, Proliferation Assay