cd137 (anti-mouse 17b5  (Bio X Cell)

Journal: eLife

Article Title: SLAMF6​ deficiency augments tumor killing and skews toward an effector phenotype revealing it as a novel T cell checkpoint

doi: 10.7554/eLife.52539

Figure Lengend Snippet: ( A ) SLAMF6 and Vβ13 expression in Pmel-1 or Pmel-1 x SLAMF6 -/- splenocytes measured by flow cytometry. ( B ) Percent CD8+, CD4, and CD19 cells in spleens from Pmel-1 or Pmel-1 x SLAMF6 -/- untreated mice. ( C ) Pmel-1, and Pmel-1 x SLAMF6 -/- CD8+ untreated splenocytes were stained with anti-CD44 and anti-CD62L. One representative experiment is shown. ( D ) Percent CD8+ cells in Pmel-1 or Pmel-1 x SLAMF6 -/- splenocytes after 7 days of in vitro activation with gp100 25-33 peptide and IL-2 (30 IU/ml). ( E ) Flow cytometry for activation markers (CD25, CD69, CD137) in Pmel-1 or Pmel-1 x SLAMF6 -/- splenocytes after 3 days of in vitro activation, as in ( D ). Median fluorescence intensity (MFI) is shown. ( F ) Expression of PD-1 in Pmel-1 or Pmel-1 x SLAMF6 -/- CD8+ T cells after 7 days of in vitro activation, as in ( D ). Median fluorescence intensity (MFI) is shown. ( G, H ) After 7 days of activation, Pmel-1 and Pmel-1 x SLAMF6 -/- CD8+ T cells were stained with anti-CD44 and anti-CD62L. CD8+ subpopulations were defined for each mouse strain. ( G ) One representative experiment and ( H ) summary of subpopulations identified by flow cytometry in five experiments is shown. EM, effector memory, CM, central memory. Student t-test. *, p<0.05, **, p<0.01, ***, p<0.001.

Article Snippet: Antibody , Monoclonal Syrian hamster anti mouse CD137 (17B5) , eBioscience, CA , 12-1371-82 , 0.2 μg/100 μl.

Techniques: Expressing, Flow Cytometry, Staining, In Vitro, Activation Assay, Fluorescence

Journal: eLife

Article Title: SLAMF6​ deficiency augments tumor killing and skews toward an effector phenotype revealing it as a novel T cell checkpoint

doi: 10.7554/eLife.52539

Figure Lengend Snippet:

Article Snippet: Antibody , Monoclonal Syrian hamster anti mouse CD137 (17B5) , eBioscience, CA , 12-1371-82 , 0.2 μg/100 μl.

Techniques: Generated, Immunohistochemistry, Sequencing, Recombinant, Enzyme-linked Immunosorbent Assay, cDNA Synthesis, Transformation Assay, Clonogenic Cell Survival Assay, Cell Isolation, Isolation, Software

Journal: Stem Cell Research & Therapy

Article Title: Mechanisms utilized by feline adipose-derived mesenchymal stem cells to inhibit T lymphocyte proliferation

doi: 10.1186/s13287-019-1300-3

Figure Lengend Snippet: ICAM-1, CD137L, and PDL-1 are all expressed on activated feline ASCs; however, CD137L and PDL-1 do not mediate MSC-T cell adhesion. Expression of a ICAM-1, b CD137L, and c PDL-1 ligands on activated feline ASCs. Gray histogram indicated background fluorescence of unstained samples. d Percentage of remaining fluorescence intensity from CMFDA-labeled PBMCs after removal of non-adherent cells from static adhesion assay after the addition of CD137/CD137L and PD-1/PDL-1 blocking antibodies. Data is normalized to 100% on a standard MLR condition for comparability. Fluorescent images of static adhesion assay demonstrating adherent lymphocytes to ASCs from e , h non-activated MLR, f , i stimulated MLR with ConA, and g , j stimulated MLR with ConA and addition of CD137/CD137L and PD-1/PDL-1 blocking antibodies respectively. Scale bar = 400 μm. Representative flow analysis images in a – c from 3 independent experiments. Data gathered in d from 5 independent experiments

Article Snippet: In some experiments, blocking antibodies to ICAM 1 (anti-human CD54, clone MEM-111, Thermo Fisher Scientific), LFA-1 (anti-human clone R7.1, eBioscience), CD137 (anti-mouse clone 17B5, Bio X Cell), CD137L (anti-mouse clone TKS-1, Bio X Cell), PD-1 (anti-human PD-1, polyclonal goat IgG, R&D systems), or PDL-1 (anti-human PDL-1, polyclonal goat IgG, R&D systems) were added to determine which ligands mediated PBMC-ASC adhesion.

Techniques: Expressing, Fluorescence, Labeling, Cell Adhesion Assay, Blocking Assay

Journal: Cell

Article Title: High Dimensional Analysis Delineates Myeloid and Lymphoid Compartment Remodeling During Successful Immune Checkpoint Cancer Therapy

doi: 10.1016/j.cell.2018.09.030

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Anti-mouse CD137 (clone 17B5) , Thermo Fisher , Cat# 16-1371-85.

Techniques: Recombinant, Mutagenesis, Sequencing, Purification, Staining, Antibody Labeling, Gene Expression, Generated, Software

Journal: Journal of Neuroinflammation

Article Title: CD137 ligand activated microglia induces oligodendrocyte apoptosis via reactive oxygen species

doi: 10.1186/1742-2094-9-173

Figure Lengend Snippet: CD137L signaling activates microglia in vitro . Microglia cells were grown on plates that had been coated with PBS (grey bars) or 10 μg/mL of Fc control protein (white bars) or 10 μg/mL of CD137-Fc protein (black bars). ( A ) Attachment and morphological changes of primary microglia were documented by photography (40× magnification) 1 h after plating. Scale bar: 20 μm. ( B ) Attachment and morphological changes of BV-2 and N9 cells and primary microglia were documented by photography (63×) 24 h after plating. Cells exposed to immobilized CD137-Fc protein developed long spiky projections* and became amoeboid cells^ with shortened protrusions # . Scale bar: 20 μm. ( C ) The concentrations of cytokines in supernatants of primary microglia were determined by ELISA at indicated time points. Depicted are means ± standard deviations of triplicate measurements. ( D ) The phagocytic capacity of the cells was determined by adding FITC-labeled latex beads for 1 h before analysis by flow cytometry. Control: Autofluorescence of the cells. Numbers above the histograms state the percentages of positive cells and mean fluorescence intensities (MFI). These experiments were repeated three times with similar results. * P <0.05; ** P <0.01.

Article Snippet: Phycoerythrin (PE)-conjugated and unconjugated anti-mouse CD137 antibody (clone 17B5) and anti-mouse CD137 ligand antibody (clone TKS-1) were obtained from eBioscience (San Diego, CA, USA).

Techniques: In Vitro, Control, Enzyme-linked Immunosorbent Assay, Labeling, Flow Cytometry, Fluorescence

Journal: Journal of Neuroinflammation

Article Title: CD137 ligand activated microglia induces oligodendrocyte apoptosis via reactive oxygen species

doi: 10.1186/1742-2094-9-173

Figure Lengend Snippet: CD137L signaling activates microglia in vivo . ( A ) CD137L is required for activation of microglia in vivo . Cortex and spinal cord tissue sections of WT and CD137L -/- mice with EAE were immunohistochemically stained with an isotype control antibody or for Iba-1 (brown). Shown in the inset is a close-up of a single Iba-1-positive microglia cell in the cortex of a WT mouse with EAE. ( B ) Quantification of Iba-1 + microglia in the spinal cords and cortices of WT and CD137L -/- mice with EAE. Evaluated were three fields from two sections each using the Metamorph NX Software. Depicted are means ± standard deviations. ( C ) CD137 is expressed in the CNS during EAE. Spinal cord of naïve WT mice and WT mice with EAE was sectioned and stained with an isotype control antibody or for CD137 (brown). ( D ) The presence of CD137L is required for oligodendrocyte apoptosis in EAE. Tissue sections from the dorsal column of the spinal cord and the white matter of the cerebellum of WT and CD137L -/- mice with EAE were stained for oligodendrocytes using a Cy3-labeled anti-Nogo-A antibody (red). Apoptosis was detected by TUNEL staining (green). Nuclei were visualized by DAPI (blue). The yellow staining results from an overlay of red and green and indicates apoptotic oligodendrocytes. Magnification: 40×. ( E ) Quantification of apoptotic oligodendrocytes in the cerebellum of WT and CD137L -/- mice with EAE. Evaluated were three fields from two sections each using the Metamorph NX Software. Depicted are means ± standard deviations.

Article Snippet: Phycoerythrin (PE)-conjugated and unconjugated anti-mouse CD137 antibody (clone 17B5) and anti-mouse CD137 ligand antibody (clone TKS-1) were obtained from eBioscience (San Diego, CA, USA).

Techniques: In Vivo, Activation Assay, Staining, Control, Software, Labeling, TUNEL Assay

Journal: Journal of Neuroinflammation

Article Title: CD137 ligand activated microglia induces oligodendrocyte apoptosis via reactive oxygen species

doi: 10.1186/1742-2094-9-173

Figure Lengend Snippet: Induction of oligodendrocyte death by CD137L-activated microglia. (A ) Expression of CD137L on the oligodendrocyte cell line OLN93 was determined by flow cytometry. Open histogram: Isotype control. Grey histogram: Anti-CD137L monoclonal antibody (clone TKS-1). ( B ) N9 and OLN93 cells were cultured for 24 h at a 1:1 ratio (1.5×10 5 each) on plates that had been coated with nothing (PBS) or 10 μg/mL of Fc control protein or 10 μg/mL of CD137-Fc protein. ( C ) The rates of OLN93 cell apoptosis in co-cultures with primary microglia from WT or CD137L -/- mice was determined 48 h after initiation of CD137L signaling by 7-AAD and Annexin V staining. Numbers in quadrants indicate percentages of cells. These experiments were repeated two to three times with similar results. * P <0.05; ** P <0.01.

Article Snippet: Phycoerythrin (PE)-conjugated and unconjugated anti-mouse CD137 antibody (clone 17B5) and anti-mouse CD137 ligand antibody (clone TKS-1) were obtained from eBioscience (San Diego, CA, USA).

Techniques: Expressing, Flow Cytometry, Control, Cell Culture, Staining

Journal: Journal of Neuroinflammation

Article Title: CD137 ligand activated microglia induces oligodendrocyte apoptosis via reactive oxygen species

doi: 10.1186/1742-2094-9-173

Figure Lengend Snippet: CD137L-activated microglia induces oligodendrocyte apoptosis via ROS. ( A ) BV-2 cells were cultured on uncoated plates (PBS) or plates coated with Fc or CD137-Fc protein for 24 h, and were then stimulated with 0.4 μg of PMA for 1 h, stained with DHR123 before production of ROS was quantified by flow cytometry. White histogram: No DHR123. The number in the panel indicates the percentage of positive cells. ( B ) BV-2 cells were co-cultured with OLN93 cells at a 1:1 ratio with or without 10,000 U/mL catalase. The rate of apoptosis of cultures was determined after 24 h by 7-AAD and Annexin V staining. Numbers in quadrants indicate percentages of cells. ( C ) The percentages of 7-AAD + OLN93 cells with and without catalase treatment of B are presented as means ± standard deviations of triplicate measurements. * P < 0.05; ** P < 0.01.

Article Snippet: Phycoerythrin (PE)-conjugated and unconjugated anti-mouse CD137 antibody (clone 17B5) and anti-mouse CD137 ligand antibody (clone TKS-1) were obtained from eBioscience (San Diego, CA, USA).

Techniques: Cell Culture, Staining, Flow Cytometry

Journal: PLoS ONE

Article Title: Regulation of Granulocyte and Macrophage Populations of Murine Bone Marrow Cells by G-CSF and CD137 Protein

doi: 10.1371/journal.pone.0015565

Figure Lengend Snippet: (A) Bone marrow cells were analyzed immediately after isolation (day 0) for cell surface marker expression by flow cytometry, or 6×10 6 bone marrow cells were cultured in T25 flasks coated with 1 ml of 10 µg/ml of Fc or CD137-Fc protein and analyzed on days 7 and 14. Black line: Isotype. Gray, filled curve: Marker-specific antibody. (B) Depicted are absolute numbers of live cells (left column) and percentages (right column) of data in (A) for indicated subpopulations at days 0, 7 and 14. (C) Fresh murine bone marrow cells, or cells which were cultured on plates coated with 10 µg/ml CD137-Fc protein for 7 days, smeared on glass slides, fixed with citrate-acetone-formaldehyde solution, and stained for non-specific esterase (black, macrophages) and for specific esterase (purple, granulocytes). Slides were counterstained with hematoxylin and mounted with cover slips. Magnification: ×630. Scale bar: 25 µm. These data are representative of three independent experiments with similar results.

Article Snippet: FITC-conjugated anti-mouse Ly-6G (clone 1A8), PE-conjugated anti-mouse Gr-1 (clone RB6-8C5), CD11b (clone M1/70), CD14 (clone Sa2-8), F4/80 (clone BM8), CD137 ligand (clone TKS-1), APC-conjugated anti-mouse CD115 (clone AFS98), Pacific blue-conjugated anti-mouse CD34 (clone RAM34), PerCP-Cy5.5-conjugated anti-mouse CD16/32 (clone 93) antibodies and respective isotype controls (rat IgG2a, rat IgG2b), and PE-conjugated hamster anti-mouse CD137 (clone 17B5) and its isotype control (Golden Syrian hamster IgG) were purchased from eBioscience (San Diego, CA).

Techniques: Isolation, Marker, Expressing, Flow Cytometry, Cell Culture, Staining

Journal: PLoS ONE

Article Title: Regulation of Granulocyte and Macrophage Populations of Murine Bone Marrow Cells by G-CSF and CD137 Protein

doi: 10.1371/journal.pone.0015565

Figure Lengend Snippet: CD11b + cells were enriched from bone marrow by MACS, and labeled with CD11b-PE and Ly6G-FITC Ab. 3×10 5 FACS-sorted CD11b + , Ly6G + cells at a density of 6×10 5 cells/ml were cultured on plates coated with 10 µg/ml Fc or CD137-Fc protein for 24 h. Cells were stained with Annexin V-PE and 7-AAD and analyzed by flow cytometry with compensation of FITC channel. PBS: Medium control. This figure is a representative of three independent experiments.

Article Snippet: FITC-conjugated anti-mouse Ly-6G (clone 1A8), PE-conjugated anti-mouse Gr-1 (clone RB6-8C5), CD11b (clone M1/70), CD14 (clone Sa2-8), F4/80 (clone BM8), CD137 ligand (clone TKS-1), APC-conjugated anti-mouse CD115 (clone AFS98), Pacific blue-conjugated anti-mouse CD34 (clone RAM34), PerCP-Cy5.5-conjugated anti-mouse CD16/32 (clone 93) antibodies and respective isotype controls (rat IgG2a, rat IgG2b), and PE-conjugated hamster anti-mouse CD137 (clone 17B5) and its isotype control (Golden Syrian hamster IgG) were purchased from eBioscience (San Diego, CA).

Techniques: Labeling, Cell Culture, Staining, Flow Cytometry, Control

Journal: PLoS ONE

Article Title: Regulation of Granulocyte and Macrophage Populations of Murine Bone Marrow Cells by G-CSF and CD137 Protein

doi: 10.1371/journal.pone.0015565

Figure Lengend Snippet: Murine bone marrow cells at a density of 10 6 cells/ml were cultured for 7 days with respective treatment. 10 µg/ml of Fc or CD137-Fc protein were pre-coated on the plates. (A) Proliferation assay: 1 ng/ml of G-CSF was added daily to indicated wells. Cells (10 5 /condition) were labeled for the last 24 h with 0.5 µCi 3 H-thymidine, and the rate of proliferation was determined with a scintillation counter (Packard, Meriden, CT). Depicted are means ± standard deviations of triplicate measurements. (B) Cell count: 10 ng/ml of G-CSF was added daily to indicated wells. Cells (2×10 6 /condition) were harvested on day 7, and cell count was determined by trypan blue staining of 4 representative microscopic fields. Depicted are means ± standard errors of seven independent experiments. * p<0.05; ** p<0.01.

Article Snippet: FITC-conjugated anti-mouse Ly-6G (clone 1A8), PE-conjugated anti-mouse Gr-1 (clone RB6-8C5), CD11b (clone M1/70), CD14 (clone Sa2-8), F4/80 (clone BM8), CD137 ligand (clone TKS-1), APC-conjugated anti-mouse CD115 (clone AFS98), Pacific blue-conjugated anti-mouse CD34 (clone RAM34), PerCP-Cy5.5-conjugated anti-mouse CD16/32 (clone 93) antibodies and respective isotype controls (rat IgG2a, rat IgG2b), and PE-conjugated hamster anti-mouse CD137 (clone 17B5) and its isotype control (Golden Syrian hamster IgG) were purchased from eBioscience (San Diego, CA).

Techniques: Cell Culture, Proliferation Assay, Labeling, Cell Counting, Staining

Journal: PLoS ONE

Article Title: Regulation of Granulocyte and Macrophage Populations of Murine Bone Marrow Cells by G-CSF and CD137 Protein

doi: 10.1371/journal.pone.0015565

Figure Lengend Snippet: 2×10 6 murine bone marrow cells at a density of 10 6 cells/ml were cultured for 7 days on plates that had been coated with 10 µg/ml of Fc or CD137-Fc protein. Where indicated, G-CSF was added daily at 10 ng/ml or other stated concentrations. The percentages of Gr-1 + cells (A) and CD14 + , F4/80 + cells (B), as well as absolute cell numbers (E), determined by flow cytometry on day 7, are depicted as means ± standard errors of 7 independent experiments. Dose responses of the combination of G-CSF and CD137 protein: Cells were treated with a fixed concentration of CD137-Fc at 10 µg/ml and varying the concentration of G-CSF at 0, 1, 10, or 100 ng/ml (C); or a fixed concentration of G-CSF at 10 ng/ml and varying the concentration of CD137-Fc at 0, 5, 10, or 20 µg/ml (D), and flow cytometry was performed for Gr-1, CD14 and F4/80 on day 7. These data are representative of two experiments with similar results. Statistical significance was determined by two tailed, paired Student's t-test. * p<0.05; ** p<0.01; n.s. not significant.

Article Snippet: FITC-conjugated anti-mouse Ly-6G (clone 1A8), PE-conjugated anti-mouse Gr-1 (clone RB6-8C5), CD11b (clone M1/70), CD14 (clone Sa2-8), F4/80 (clone BM8), CD137 ligand (clone TKS-1), APC-conjugated anti-mouse CD115 (clone AFS98), Pacific blue-conjugated anti-mouse CD34 (clone RAM34), PerCP-Cy5.5-conjugated anti-mouse CD16/32 (clone 93) antibodies and respective isotype controls (rat IgG2a, rat IgG2b), and PE-conjugated hamster anti-mouse CD137 (clone 17B5) and its isotype control (Golden Syrian hamster IgG) were purchased from eBioscience (San Diego, CA).

Techniques: Cell Culture, Flow Cytometry, Concentration Assay, Two Tailed Test

Journal: PLoS ONE

Article Title: Regulation of Granulocyte and Macrophage Populations of Murine Bone Marrow Cells by G-CSF and CD137 Protein

doi: 10.1371/journal.pone.0015565

Figure Lengend Snippet: 5×10 5 lin − , c-kit + cells at a density of 5×10 5 cells/ml were cultured on plates that had been coated with 10 µg/ml of Fc or CD137-Fc protein or that were not coated (PBS, medium control). Where indicated, G-CSF was added daily at 10 ng/ml. The percentages (A) and absolute numbers (B) of CD11b + , Ly6G − and CD11b + , Ly6G + cells, and the percentages (C) and absolute numbers (D) of CD115 + , MPO − and CD115 − , MPO + cells were determined by flow cytometry on day 7. (A) and (C) are representatives of five and two experiments, respectively, (B) and (D) are depicted as means ± standard errors of five and two independent experiments, respectively. * p<0.05.

Article Snippet: FITC-conjugated anti-mouse Ly-6G (clone 1A8), PE-conjugated anti-mouse Gr-1 (clone RB6-8C5), CD11b (clone M1/70), CD14 (clone Sa2-8), F4/80 (clone BM8), CD137 ligand (clone TKS-1), APC-conjugated anti-mouse CD115 (clone AFS98), Pacific blue-conjugated anti-mouse CD34 (clone RAM34), PerCP-Cy5.5-conjugated anti-mouse CD16/32 (clone 93) antibodies and respective isotype controls (rat IgG2a, rat IgG2b), and PE-conjugated hamster anti-mouse CD137 (clone 17B5) and its isotype control (Golden Syrian hamster IgG) were purchased from eBioscience (San Diego, CA).

Techniques: Cell Culture, Control, Flow Cytometry

Journal: PLoS ONE

Article Title: Regulation of Granulocyte and Macrophage Populations of Murine Bone Marrow Cells by G-CSF and CD137 Protein

doi: 10.1371/journal.pone.0015565

Figure Lengend Snippet: (A) Lin − , c-kit + cells (boxed area of left panel) were sorted for CD34 + , CD16/32 high (GMP) and CD34 + , CD16/32 low (CMP) cells (right panel). (B) 5×10 4 CMP or GMP at a density of 2.5×10 5 cells/ml were cultured for 7 days on plates that had been coated with 10 µg/ml of Fc or CD137-Fc protein or that were not coated (PBS, medium control). Photographs were taken on day 7 at magnification of ×200. Scale bar = 50 µm. Please note that the small round particles are cell debris as evidenced by their small size and the holes in the membranes. (C) The percentages of CD11b + , Ly6G − and CD11b + , Ly6G + cells of (B) were determined by flow cytometry.

Article Snippet: FITC-conjugated anti-mouse Ly-6G (clone 1A8), PE-conjugated anti-mouse Gr-1 (clone RB6-8C5), CD11b (clone M1/70), CD14 (clone Sa2-8), F4/80 (clone BM8), CD137 ligand (clone TKS-1), APC-conjugated anti-mouse CD115 (clone AFS98), Pacific blue-conjugated anti-mouse CD34 (clone RAM34), PerCP-Cy5.5-conjugated anti-mouse CD16/32 (clone 93) antibodies and respective isotype controls (rat IgG2a, rat IgG2b), and PE-conjugated hamster anti-mouse CD137 (clone 17B5) and its isotype control (Golden Syrian hamster IgG) were purchased from eBioscience (San Diego, CA).

Techniques: Cell Culture, Control, Flow Cytometry