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4 1bb agonist mab  (R&D Systems)


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    R&D Systems 4 1bb agonist mab
    4 1bb Agonist Mab, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/agonist+4+1bb+mab/Mouse+4-1BB%2FTNFRSF9%2FCD137+Antibody/us12371499-2425-20-25
    Average 93 stars, based on 4 article reviews
    4 1bb agonist mab - by Bioz Stars, 2026-09
    93/100 stars

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    Related Articles

    Activation Assay:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).. Agonist 4-1BB mAb (10 µg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.. Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.

    Western Blot:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).. Agonist 4-1BB mAb (10 µg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.. Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.

    Real-time Polymerase Chain Reaction:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).. Agonist 4-1BB mAb (10 µg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.. Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.

    Expressing:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).. Agonist 4-1BB mAb (10 µg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.. Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.

    Enzyme-linked Immunosorbent Assay:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).. Agonist 4-1BB mAb (10 µg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.. Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.

    Transfection:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).. Agonist 4-1BB mAb (10 µg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.. Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.

    Saline:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).. Agonist 4-1BB mAb (10 µg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.. Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.

    Isolation:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).. Agonist 4-1BB mAb (10 µg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.. Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.

    Inhibition:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).. Agonist 4-1BB mAb (10 µg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.. Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.

    Suspension:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).. Agonist 4-1BB mAb (10 µg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.. Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.

    Control:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).. Agonist 4-1BB mAb (10 µg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.. Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.

    Injection:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).. Agonist 4-1BB mAb (10 µg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.. Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.

    Immunohistochemical staining:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).. Agonist 4-1BB mAb (10 µg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.. Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.

    Staining:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).To examine the expression of IL-1β, MH-S cells were pretreated with lipopolysaccharide (LPS, 25 ng/mL, Sigma-Aldrich Chemical Company) for 3 h (Figure J).. Agonist 4-1BB mAb (10 µg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.. Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.Isotype IgG (10 µg/mL, Sino Biological Inc.) was used as a control.



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    CD4+ TEα cells were adoptively transferred into High or Low mice on day 0, followed by immunotherapy with either dual costimulation (DCo) or control IgG on day 1. Splenocytes were harvested on day 6 for analysis via flow cytometry. Cells are gated on forward scatter (FSC) and side scatter (SSC).

    Journal: Cell reports

    Article Title: Costimulation Induces CD4 T Cell Antitumor Immunity via an Innate-like Mechanism

    doi: 10.1016/j.celrep.2019.04.016

    Figure Lengend Snippet: CD4+ TEα cells were adoptively transferred into High or Low mice on day 0, followed by immunotherapy with either dual costimulation (DCo) or control IgG on day 1. Splenocytes were harvested on day 6 for analysis via flow cytometry. Cells are gated on forward scatter (FSC) and side scatter (SSC).

    Article Snippet: Approximately 2.5 × 10 4 TEα cells were i.v. transferred into High or Low recipient mice and the following day mice were injected i.p. with 50μg of anti-CD134/OX40 and 25 μg of anti-CD137/4–1BB agonist mAbs, or with 75 μg of rat IgG isotype control antibody (Bio X Cell, West Lebanon, NH) similar to our previous work ( Lee et al., 2004 ; Mittal et al., 2015 ). also included B6 recipients treated the following day with 100 μg Eα peptide 52–68 (Invivogen, San Diego, CA) i.p. together with 50 μg of anti-CD134 (clone OX86, IgG1) and 25 mg anti-CD137 (clone 3H3, IgG2a) similar to our previous work ( McAleer et al., 2010 ).

    Techniques: Flow Cytometry

    Adoptive transfer of TEa cells into High or Low mice on day 0 followed by DCo or IgG immunotherapy on day 1. Flow cytometry of day 6 splenocytes was analyzed. Cells are gated on FSC, SSC, CD4+, Vα2+, and Vβ6+.

    Journal: Cell reports

    Article Title: Costimulation Induces CD4 T Cell Antitumor Immunity via an Innate-like Mechanism

    doi: 10.1016/j.celrep.2019.04.016

    Figure Lengend Snippet: Adoptive transfer of TEa cells into High or Low mice on day 0 followed by DCo or IgG immunotherapy on day 1. Flow cytometry of day 6 splenocytes was analyzed. Cells are gated on FSC, SSC, CD4+, Vα2+, and Vβ6+.

    Article Snippet: Approximately 2.5 × 10 4 TEα cells were i.v. transferred into High or Low recipient mice and the following day mice were injected i.p. with 50μg of anti-CD134/OX40 and 25 μg of anti-CD137/4–1BB agonist mAbs, or with 75 μg of rat IgG isotype control antibody (Bio X Cell, West Lebanon, NH) similar to our previous work ( Lee et al., 2004 ; Mittal et al., 2015 ). also included B6 recipients treated the following day with 100 μg Eα peptide 52–68 (Invivogen, San Diego, CA) i.p. together with 50 μg of anti-CD134 (clone OX86, IgG1) and 25 mg anti-CD137 (clone 3H3, IgG2a) similar to our previous work ( McAleer et al., 2010 ).

    Techniques: Adoptive Transfer Assay, Flow Cytometry

    Adoptive transfer of TEα cells into High and Low mice on day 0 was followed by DCo or IgG immunotherapy on day 1. Day 6 splenocytes were stimulated ex vivo with 5 μg/mL Eα -peptide, 1X PMA/I (P+I), or media control for 4 h with brefeldin A (BFA), followed by intracellular staining. Cells were additionally treated with control or YAe blocking antibody and are gated on FSC, SSC, CD4+, Vα2+, and Vβ6+.

    Journal: Cell reports

    Article Title: Costimulation Induces CD4 T Cell Antitumor Immunity via an Innate-like Mechanism

    doi: 10.1016/j.celrep.2019.04.016

    Figure Lengend Snippet: Adoptive transfer of TEα cells into High and Low mice on day 0 was followed by DCo or IgG immunotherapy on day 1. Day 6 splenocytes were stimulated ex vivo with 5 μg/mL Eα -peptide, 1X PMA/I (P+I), or media control for 4 h with brefeldin A (BFA), followed by intracellular staining. Cells were additionally treated with control or YAe blocking antibody and are gated on FSC, SSC, CD4+, Vα2+, and Vβ6+.

    Article Snippet: Approximately 2.5 × 10 4 TEα cells were i.v. transferred into High or Low recipient mice and the following day mice were injected i.p. with 50μg of anti-CD134/OX40 and 25 μg of anti-CD137/4–1BB agonist mAbs, or with 75 μg of rat IgG isotype control antibody (Bio X Cell, West Lebanon, NH) similar to our previous work ( Lee et al., 2004 ; Mittal et al., 2015 ). also included B6 recipients treated the following day with 100 μg Eα peptide 52–68 (Invivogen, San Diego, CA) i.p. together with 50 μg of anti-CD134 (clone OX86, IgG1) and 25 mg anti-CD137 (clone 3H3, IgG2a) similar to our previous work ( McAleer et al., 2010 ).

    Techniques: Adoptive Transfer Assay, Ex Vivo, Staining, Blocking Assay

    High and Low mice were treated with DCo or IgG 1 day after TEα adoptive transfer. Day 6 splenocytes were analyzed via flow cytometry. Cells are gated on FSC, SSC, and CD4+ cells; FSC, SSC, CD4+, Vα2+ and Vβ6+ (TEα) cells; or FSC, SSC, CD4+, and Vα2/Vβ6 negative or single positive (non-Vα2 Vβ6 cells/non-TEα).

    Journal: Cell reports

    Article Title: Costimulation Induces CD4 T Cell Antitumor Immunity via an Innate-like Mechanism

    doi: 10.1016/j.celrep.2019.04.016

    Figure Lengend Snippet: High and Low mice were treated with DCo or IgG 1 day after TEα adoptive transfer. Day 6 splenocytes were analyzed via flow cytometry. Cells are gated on FSC, SSC, and CD4+ cells; FSC, SSC, CD4+, Vα2+ and Vβ6+ (TEα) cells; or FSC, SSC, CD4+, and Vα2/Vβ6 negative or single positive (non-Vα2 Vβ6 cells/non-TEα).

    Article Snippet: Approximately 2.5 × 10 4 TEα cells were i.v. transferred into High or Low recipient mice and the following day mice were injected i.p. with 50μg of anti-CD134/OX40 and 25 μg of anti-CD137/4–1BB agonist mAbs, or with 75 μg of rat IgG isotype control antibody (Bio X Cell, West Lebanon, NH) similar to our previous work ( Lee et al., 2004 ; Mittal et al., 2015 ). also included B6 recipients treated the following day with 100 μg Eα peptide 52–68 (Invivogen, San Diego, CA) i.p. together with 50 μg of anti-CD134 (clone OX86, IgG1) and 25 mg anti-CD137 (clone 3H3, IgG2a) similar to our previous work ( McAleer et al., 2010 ).

    Techniques: Adoptive Transfer Assay, Flow Cytometry

    B6 recipients bearing day 3 established B16-F10 tumors received dual costimulation along with 3.3 × 105 retransferred day 6 FACS sorted TEα Thy1.1+ TCR Tg CD4 T cells recovered from dual costimulated High recipient mice. These tumor-bearing mice were divided into 4 groups that received (1) a single intraperitoneal (i.p.) injection of 250 μg soluble Eα -peptide to activate the re-transferred specific CD4 helper T cells plus 1 mg intratumorally injected IL-36β on days 5, 7, 9, and 11 (+ Ag + IL-36β); (2) Eα -peptide plus intratumoral PBS injections (+ Ag [C0] IL-36β); (3) control rat IgG plus intratumoral IL-36b injections ([C0] Ag + IL-36β); and (4) control rat IgG plus intratumoral PBS injections ([C0] Ag [C0] IL-36b). n = 9–10 mice/group.

    Journal: Cell reports

    Article Title: Costimulation Induces CD4 T Cell Antitumor Immunity via an Innate-like Mechanism

    doi: 10.1016/j.celrep.2019.04.016

    Figure Lengend Snippet: B6 recipients bearing day 3 established B16-F10 tumors received dual costimulation along with 3.3 × 105 retransferred day 6 FACS sorted TEα Thy1.1+ TCR Tg CD4 T cells recovered from dual costimulated High recipient mice. These tumor-bearing mice were divided into 4 groups that received (1) a single intraperitoneal (i.p.) injection of 250 μg soluble Eα -peptide to activate the re-transferred specific CD4 helper T cells plus 1 mg intratumorally injected IL-36β on days 5, 7, 9, and 11 (+ Ag + IL-36β); (2) Eα -peptide plus intratumoral PBS injections (+ Ag [C0] IL-36β); (3) control rat IgG plus intratumoral IL-36b injections ([C0] Ag + IL-36β); and (4) control rat IgG plus intratumoral PBS injections ([C0] Ag [C0] IL-36b). n = 9–10 mice/group.

    Article Snippet: Approximately 2.5 × 10 4 TEα cells were i.v. transferred into High or Low recipient mice and the following day mice were injected i.p. with 50μg of anti-CD134/OX40 and 25 μg of anti-CD137/4–1BB agonist mAbs, or with 75 μg of rat IgG isotype control antibody (Bio X Cell, West Lebanon, NH) similar to our previous work ( Lee et al., 2004 ; Mittal et al., 2015 ). also included B6 recipients treated the following day with 100 μg Eα peptide 52–68 (Invivogen, San Diego, CA) i.p. together with 50 μg of anti-CD134 (clone OX86, IgG1) and 25 mg anti-CD137 (clone 3H3, IgG2a) similar to our previous work ( McAleer et al., 2010 ).

    Techniques: Injection

    KEY RESOURCES TABLE

    Journal: Cell reports

    Article Title: Costimulation Induces CD4 T Cell Antitumor Immunity via an Innate-like Mechanism

    doi: 10.1016/j.celrep.2019.04.016

    Figure Lengend Snippet: KEY RESOURCES TABLE

    Article Snippet: Approximately 2.5 × 10 4 TEα cells were i.v. transferred into High or Low recipient mice and the following day mice were injected i.p. with 50μg of anti-CD134/OX40 and 25 μg of anti-CD137/4–1BB agonist mAbs, or with 75 μg of rat IgG isotype control antibody (Bio X Cell, West Lebanon, NH) similar to our previous work ( Lee et al., 2004 ; Mittal et al., 2015 ). also included B6 recipients treated the following day with 100 μg Eα peptide 52–68 (Invivogen, San Diego, CA) i.p. together with 50 μg of anti-CD134 (clone OX86, IgG1) and 25 mg anti-CD137 (clone 3H3, IgG2a) similar to our previous work ( McAleer et al., 2010 ).

    Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Transgenic Assay, Software

    Sequences.

    Journal: Frontiers in Immunology

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice

    doi: 10.3389/fimmu.2018.01848

    Figure Lengend Snippet: Sequences.

    Article Snippet: Agonist 4-1BB mAb (10 μg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.

    Techniques:

    Activation of 4-1BB signaling promotes the secretion of pro-fibrotic mediators by MH-S cells. MH-S cells treated with or without agonist 4-1BB mAb (10 µg/mL) or IgG (10 µg/mL) for 2 h prior to exposure to crystalline silica (50 µg/cm 2 ) for 12 h. (A) Western blots analysis of ASK-1 and downstream mitogen-activated protein kinase proteins (p38 and JNK/stress activated protein kinase) and their phosphorylated forms. (B–D) The levels of phospho-ASK1, phospho-p38, and phospho-JNK were normalized to those of β-actin ( n = 3). (E) Western blots analysis of IκBα and phospho-IκBα. (F) The level of phospho-IκBα was normalized to those of β-actin ( n = 3). (G–I) Real-time polymerase chain reaction analysis of MMP12, MMP9, and monocyte chemoattractant protein-1 mRNA expression ( n = 4). (J–L) ELISA analysis was used to quantify the secretion of IL-1β, IL-6 and tumor necrosis factor-α ( n = 4). The results were representative of three independent experiments. Results were graphed as the mean ± SEM (* p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001).

    Journal: Frontiers in Immunology

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice

    doi: 10.3389/fimmu.2018.01848

    Figure Lengend Snippet: Activation of 4-1BB signaling promotes the secretion of pro-fibrotic mediators by MH-S cells. MH-S cells treated with or without agonist 4-1BB mAb (10 µg/mL) or IgG (10 µg/mL) for 2 h prior to exposure to crystalline silica (50 µg/cm 2 ) for 12 h. (A) Western blots analysis of ASK-1 and downstream mitogen-activated protein kinase proteins (p38 and JNK/stress activated protein kinase) and their phosphorylated forms. (B–D) The levels of phospho-ASK1, phospho-p38, and phospho-JNK were normalized to those of β-actin ( n = 3). (E) Western blots analysis of IκBα and phospho-IκBα. (F) The level of phospho-IκBα was normalized to those of β-actin ( n = 3). (G–I) Real-time polymerase chain reaction analysis of MMP12, MMP9, and monocyte chemoattractant protein-1 mRNA expression ( n = 4). (J–L) ELISA analysis was used to quantify the secretion of IL-1β, IL-6 and tumor necrosis factor-α ( n = 4). The results were representative of three independent experiments. Results were graphed as the mean ± SEM (* p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001).

    Article Snippet: Agonist 4-1BB mAb (10 μg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.

    Techniques: Activation Assay, Western Blot, Real-time Polymerase Chain Reaction, Expressing, Enzyme-linked Immunosorbent Assay

    Blockade of 4-1BB signaling attenuates the secretion of pro-fibrotic mediators by MH-S cells. Transfected (Len-cont. and sh-4-1BB) MH-S cells were treated with or without NQDI 1 (10 µM), 4-1BBIg (10 µg/mL), or IgG1 (10 µg/mL) for 2 h, then exposed to crystalline silica (50 µg/cm 2 ) for 12 h. (A) Western blots analysis of ASK-1 and downstream mitogen-activated protein kinase proteins (p38 and JNK/stress activated protein kinase) and their phosphorylated forms. (B–D) Levels of phospho-ASK1, phospho-p38, and phospho-JNK were normalized to those of β-actin ( n = 3). (E,G) Western blots analysis of IκBα and phospho-IκBα. (F,H) The level of phospho-IκBα was normalized to those of β-actin ( n = 3). (I–K) The expressions of MMP12, MMP9, and monocyte chemoattractant protein-1 were detected by real-time polymerase chain reaction analysis ( n = 4). ELISA analysis of cytokines in the culture supernatants. (L) IL-1β, (M) IL-6, (N) tumor necrosis factor-α ( n = 4). Data were shown as mean ± SEM (* p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; ns, not significant). The data were representative of three independent experiments.

    Journal: Frontiers in Immunology

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice

    doi: 10.3389/fimmu.2018.01848

    Figure Lengend Snippet: Blockade of 4-1BB signaling attenuates the secretion of pro-fibrotic mediators by MH-S cells. Transfected (Len-cont. and sh-4-1BB) MH-S cells were treated with or without NQDI 1 (10 µM), 4-1BBIg (10 µg/mL), or IgG1 (10 µg/mL) for 2 h, then exposed to crystalline silica (50 µg/cm 2 ) for 12 h. (A) Western blots analysis of ASK-1 and downstream mitogen-activated protein kinase proteins (p38 and JNK/stress activated protein kinase) and their phosphorylated forms. (B–D) Levels of phospho-ASK1, phospho-p38, and phospho-JNK were normalized to those of β-actin ( n = 3). (E,G) Western blots analysis of IκBα and phospho-IκBα. (F,H) The level of phospho-IκBα was normalized to those of β-actin ( n = 3). (I–K) The expressions of MMP12, MMP9, and monocyte chemoattractant protein-1 were detected by real-time polymerase chain reaction analysis ( n = 4). ELISA analysis of cytokines in the culture supernatants. (L) IL-1β, (M) IL-6, (N) tumor necrosis factor-α ( n = 4). Data were shown as mean ± SEM (* p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; ns, not significant). The data were representative of three independent experiments.

    Article Snippet: Agonist 4-1BB mAb (10 μg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.

    Techniques: Transfection, Western Blot, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    Expression of 4-1BB (CD137) and 4-1BBL (CD137L) on pulmonary macrophages. After 7-days exposure to crystalline silica, mice were sacrificed. The lungs were prepared as single-cell suspensions for flow cytometric analyses ( n = 3–4). (A) Representative plots of flow cytometric analyses for 4-1BB and 4-1BBL on alveolar macrophages (AMs) and interstitial macrophages (IMs). (B,C) The percentage of AMs expressing 4-1BB and 4-1BBL. (D,E) The frequency of AMs and IMs in CD45+ cells from the lungs. (F,G) The percentage of IMs expressing 4-1BB and 4-1BBL. The experiments were performed twice with similar results. Data were shown as mean ± SEM (* p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; ns, not significant).

    Journal: Frontiers in Immunology

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice

    doi: 10.3389/fimmu.2018.01848

    Figure Lengend Snippet: Expression of 4-1BB (CD137) and 4-1BBL (CD137L) on pulmonary macrophages. After 7-days exposure to crystalline silica, mice were sacrificed. The lungs were prepared as single-cell suspensions for flow cytometric analyses ( n = 3–4). (A) Representative plots of flow cytometric analyses for 4-1BB and 4-1BBL on alveolar macrophages (AMs) and interstitial macrophages (IMs). (B,C) The percentage of AMs expressing 4-1BB and 4-1BBL. (D,E) The frequency of AMs and IMs in CD45+ cells from the lungs. (F,G) The percentage of IMs expressing 4-1BB and 4-1BBL. The experiments were performed twice with similar results. Data were shown as mean ± SEM (* p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; ns, not significant).

    Article Snippet: Agonist 4-1BB mAb (10 μg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.

    Techniques: Expressing

    Expression of 4-1BB on CD4+ T cells. (A) Representative plots of flow cytometric analyses for 4-1BB on effector and naïve T cells. (B) The percentage of effector T cells expressing 4-1BB. (C,D) The frequency of effector and naïve T cells in CD4+ T cells from the lungs. (E) The percentage of naïve T cells expressing 4-1BB. The experiments were performed twice with similar results. Data were shown as mean ± SEM (* p ≤ 0.05; ** p ≤ 0.01; ns, not significant).

    Journal: Frontiers in Immunology

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice

    doi: 10.3389/fimmu.2018.01848

    Figure Lengend Snippet: Expression of 4-1BB on CD4+ T cells. (A) Representative plots of flow cytometric analyses for 4-1BB on effector and naïve T cells. (B) The percentage of effector T cells expressing 4-1BB. (C,D) The frequency of effector and naïve T cells in CD4+ T cells from the lungs. (E) The percentage of naïve T cells expressing 4-1BB. The experiments were performed twice with similar results. Data were shown as mean ± SEM (* p ≤ 0.05; ** p ≤ 0.01; ns, not significant).

    Article Snippet: Agonist 4-1BB mAb (10 μg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.

    Techniques: Expressing

    4-1BB expression on the mouse alveolar macrophage cell line, MH-S. MH-S cells were treated with crystalline silica (50 µg/cm 2 ) or saline for 12 h. (A,B) The percentage of MH-S cells expressing 4-1BB ( n = 4). (C) Western blot analysis of 4-1BB protein from whole cell lysates. (D) Quantification of the 4-1BB protein level relative to that of β-actin is shown ( n = 3). (E) Total RNA was isolated to analyze 4-1BB mRNA expression ( n = 4) relative to GAPDH. The data were representative of three independent experiments. Data were expressed as mean ± SEM (*** p ≤ 0.001).

    Journal: Frontiers in Immunology

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice

    doi: 10.3389/fimmu.2018.01848

    Figure Lengend Snippet: 4-1BB expression on the mouse alveolar macrophage cell line, MH-S. MH-S cells were treated with crystalline silica (50 µg/cm 2 ) or saline for 12 h. (A,B) The percentage of MH-S cells expressing 4-1BB ( n = 4). (C) Western blot analysis of 4-1BB protein from whole cell lysates. (D) Quantification of the 4-1BB protein level relative to that of β-actin is shown ( n = 3). (E) Total RNA was isolated to analyze 4-1BB mRNA expression ( n = 4) relative to GAPDH. The data were representative of three independent experiments. Data were expressed as mean ± SEM (*** p ≤ 0.001).

    Article Snippet: Agonist 4-1BB mAb (10 μg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.

    Techniques: Expressing, Saline, Western Blot, Isolation

    The secretion of pro-fibrotic mediators is reduced in the lungs from crystalline silica (CS)-injured mice, upon inhibition of 4-1BB signaling. C57BL/6 mice were administered a CS suspension or saline, respectively; 4-1BBIg or isotype control (IgG1) were injected intraperitoneally (i.p.; n = 3–4). (A–D) Quantification of MMP9 and MMP12 protein levels by western blot, which were normalized to those of β-actin in lungs. Shown as bar graph. (E–H) ELISA analysis of cytokines in lung tissues. (E) IL-1β, (F) IL-6, (G) tumor necrosis factor-α, (H) monocyte chemoattractant protein-1. Experiments were performed three times. C57BL/6 mice were administered a CS suspension or saline, respectively; NQDI 1 or isotype control were injected i.p. ( n = 10). (I) Immunohistochemical staining of paraffin-embedded lung tissue sections at 7 and 56 days showed CD68, MMP9, and MMP12 expression. Nuclei were stained by hematoxylin (blue). (J–L) Identification of MMP9 and MMP12 protein levels in mouse lung tissues at 7 and 56 days by western blot. The levels of MMP9 and MMP12 were normalized to those of β-actin. (M) Representative images for the immunohistochemical staining of collagen I in paraffin-embedded lung tissue sections 56 days after CS instillation. Nuclei were stained by hematoxylin (blue). (I,M) Scale bar, 50 µm. Experiments were performed three times. Data are shown as mean ± SEM (* p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; ns, not significant).

    Journal: Frontiers in Immunology

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice

    doi: 10.3389/fimmu.2018.01848

    Figure Lengend Snippet: The secretion of pro-fibrotic mediators is reduced in the lungs from crystalline silica (CS)-injured mice, upon inhibition of 4-1BB signaling. C57BL/6 mice were administered a CS suspension or saline, respectively; 4-1BBIg or isotype control (IgG1) were injected intraperitoneally (i.p.; n = 3–4). (A–D) Quantification of MMP9 and MMP12 protein levels by western blot, which were normalized to those of β-actin in lungs. Shown as bar graph. (E–H) ELISA analysis of cytokines in lung tissues. (E) IL-1β, (F) IL-6, (G) tumor necrosis factor-α, (H) monocyte chemoattractant protein-1. Experiments were performed three times. C57BL/6 mice were administered a CS suspension or saline, respectively; NQDI 1 or isotype control were injected i.p. ( n = 10). (I) Immunohistochemical staining of paraffin-embedded lung tissue sections at 7 and 56 days showed CD68, MMP9, and MMP12 expression. Nuclei were stained by hematoxylin (blue). (J–L) Identification of MMP9 and MMP12 protein levels in mouse lung tissues at 7 and 56 days by western blot. The levels of MMP9 and MMP12 were normalized to those of β-actin. (M) Representative images for the immunohistochemical staining of collagen I in paraffin-embedded lung tissue sections 56 days after CS instillation. Nuclei were stained by hematoxylin (blue). (I,M) Scale bar, 50 µm. Experiments were performed three times. Data are shown as mean ± SEM (* p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; ns, not significant).

    Article Snippet: Agonist 4-1BB mAb (10 μg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.

    Techniques: Inhibition, Suspension, Saline, Control, Injection, Western Blot, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Staining, Expressing

    A model for alveolar macrophages (AMs) expressing 4-1BB in the regulation of pulmonary fibrosis through secreting pro-fibrotic mediators. The expression of 4-1BB increases in AMs in response to crystalline silica, which leads to elevated secretion of pro-inflammatory and pro-fibrotic cytokines, chemokines, and MMPs. These pro-fibrotic mediators promote pulmonary alveoli injury, the accumulation of monocytes, lymphocytes, and fibrocytes, and collagen deposition, resulting in pulmonary fibrosis.

    Journal: Frontiers in Immunology

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice

    doi: 10.3389/fimmu.2018.01848

    Figure Lengend Snippet: A model for alveolar macrophages (AMs) expressing 4-1BB in the regulation of pulmonary fibrosis through secreting pro-fibrotic mediators. The expression of 4-1BB increases in AMs in response to crystalline silica, which leads to elevated secretion of pro-inflammatory and pro-fibrotic cytokines, chemokines, and MMPs. These pro-fibrotic mediators promote pulmonary alveoli injury, the accumulation of monocytes, lymphocytes, and fibrocytes, and collagen deposition, resulting in pulmonary fibrosis.

    Article Snippet: Agonist 4-1BB mAb (10 μg/mL; cat: MAB9371, R&D Systems, Minneapolis, MN, USA) ( ) was used to detect the effect of 4-1BB signaling on macrophages 2 h before CS exposure.

    Techniques: Expressing