Journal: Cell

Article Title: Therapeutic potential of co-signaling receptor modulation in hepatitis B

doi: 10.1016/j.cell.2024.05.038

Figure Lengend Snippet:

Article Snippet: InVivoPlus anti-mouse CD137 (4-1BB) , Bio X Cell , Cat# BE0239; RRID: AB_2687721.

Techniques: Purification, Virus, Recombinant, Staining, Saline, Fluorsave, Sequencing, DNA Labeling, Cell Isolation, Sample Prep, Reverse Transcription, Software, Microscopy

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:

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

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

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

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

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

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

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

Journal: Signal transduction and targeted therapy

Article Title: Dual targeting PD-L1 and 4-1BB to overcome dendritic cell-mediated lenalidomide resistance in follicular lymphoma.

doi: 10.1038/s41392-024-02105-7

Figure Lengend Snippet: Fig. 4 PU.1 modulated PD-1/PD-L1 and 4-1BB/4-1BBL-mediated FL cell interaction with cDC1. a Immune checkpoints with differential expression between the cDC1 high and low groups identified by RNA sequencing. b Correlation between PU.1 and PD-L1, as well as PU.1 and 4-1BBL expression calculated by Pearson correlation coefficient analysis. c Relative luciferase activities in PD-L1 WT or PD-L1 MUT transfected HEK-293T cells, SC1 cells, DOHH2 cells were detected. Mean with SD was presented (n = 3). d Relative luciferase activities in 4-1BBL WT or 4-1BBL MUT transfected HEK-293T cells, SC1 cells, DOHH2 cells were detected. Mean with SD was presented (n = 3). e Western blot of PU.1, PD-L1, and 4-1BBL expression in the SpCas9-PU.1 transfected SC1 cells and pGMLV-PU.1 transfected DOHH2 cells

Article Snippet: For PD-L1 and 4-1BB antibody treatment, each mouse received an injection of 200 μg anti-mouse 4-1BB agonistic antibody (BE0296, Bioxcell, Lebanon, USA) and anti-mouse PD-L1 antibody (BE0101, Bioxcell) via the tail vein, three times each week for 2 weeks.

Techniques: Quantitative Proteomics, RNA Sequencing, Expressing, Luciferase, Transfection, Western Blot

Journal: Signal transduction and targeted therapy

Article Title: Dual targeting PD-L1 and 4-1BB to overcome dendritic cell-mediated lenalidomide resistance in follicular lymphoma.

doi: 10.1038/s41392-024-02105-7

Figure Lengend Snippet: Fig. 5 Dual targeting PD-L1 and 4-1BB counteracted PU.1-mediated cDC1 alterations in vitro. a Cell growth of the SpCas9-PU.1-transfected SC1 co-culture system, treated with or without the bi-specific PD-L1/4-1BB antibody assessed by MTT assay. Mean with SD was presented (n = 5). b cDC1 absolute cell numbers in the SpCas9-PU.1 transfected SC1 co-culture system treated with or without bi-specific PD-L1/4-1BB antibody assessed by flow cytometry. Mean with SD was presented (n = 3). c cDC1 maturation in the SpCas9-PU.1 transfected SC1 co-culture system treated with or without bi-specific PD-L1/4-1BB antibody assessed by flow cytometry. Mean with SD was presented (n = 3). d cDC1 endocytic activity in the SpCas9-PU.1 transfected SC1 co-culture system treated with or without bi-specific PD-L1/4-1BB antibody assessed by flow cytometry. Mean with SD was presented (n = 3). e LC3B expression in the SpCas9-PU.1 transfected SC1 co-culture system treated with or without bi-specific PD-L1/4-1BB antibody. Mean with SD was presented (n = 3). f Typical autophagosomes in the SpCas9-PU.1-transfected SC1 co-culture system, with or without bi-specific PD-L1/4-1BB antibody treatment assessed by transmission electron microscope. Cells were counted from randomly selected fields (n = 5). Mean with SD was presented

Article Snippet: For PD-L1 and 4-1BB antibody treatment, each mouse received an injection of 200 μg anti-mouse 4-1BB agonistic antibody (BE0296, Bioxcell, Lebanon, USA) and anti-mouse PD-L1 antibody (BE0101, Bioxcell) via the tail vein, three times each week for 2 weeks.

Techniques: In Vitro, Transfection, Co-Culture Assay, MTT Assay, Cytometry, Activity Assay, Expressing, Transmission Assay, Microscopy

Journal: Signal transduction and targeted therapy

Article Title: Dual targeting PD-L1 and 4-1BB to overcome dendritic cell-mediated lenalidomide resistance in follicular lymphoma.

doi: 10.1038/s41392-024-02105-7

Figure Lengend Snippet: Fig. 6 Combined treatment of PD-L1 and 4-1BB antibody exhibited in vivo anti-lymphoma activity on PU.1-altered murine xenograft model. a SpCas9-scramble and SpCas9-PU.1 cells were injected subcutaneously treated with or without lenalidomide, or co-treated with PD-L1 and 4-1BB antibody. Tumor size was measured. Mean with SD was presented (n = 3). b Standardized uptake value intensity in SpCas9-scramble group and SpCas9-PU.1 group treated with or without lenalidomide, or co-treated with PD-L1 and 4-1BB antibody assessed by Micro PET-CT. c, d cDC1 maturation in SpCas9-scramble group and SpCas9-PU.1 group treated with or without lenalidomide, or co-treated with PD-L1 and 4-1BB antibody assessed by flow cytometry. Mean with SD was presented (n = 3). e Flow cytometry analysis of cDC1 endocytic activity in SpCas9-scramble group and SpCas9-PU.1 group treated with or without lenalidomide, or co-treated with PD-L1 and 4-1BB antibody. Mean with SD was presented (n = 3). f Typical autophagosomes in the SpCas9-scramble group and SpCas9-PU.1 group, with or without lenalidomide treatment, or co-treated with PD-L1 and 4-1BB antibody assessed by transmission electron microscopy. Cells were counted from randomly selected fields (n = 5). Mean with SD was presented

Article Snippet: For PD-L1 and 4-1BB antibody treatment, each mouse received an injection of 200 μg anti-mouse 4-1BB agonistic antibody (BE0296, Bioxcell, Lebanon, USA) and anti-mouse PD-L1 antibody (BE0101, Bioxcell) via the tail vein, three times each week for 2 weeks.

Techniques: In Vivo, Activity Assay, Injection, Micro-PET, Cytometry, Flow Cytometry, Transmission Assay, Electron Microscopy

Journal: Nutrition & Metabolism

Article Title: Evaluation of markers of beige adipocytes in white adipose tissue of the mouse

doi: 10.1186/s12986-016-0081-2

Figure Lengend Snippet: List or thermoregulatory or beige markers employed in this study and their function

Article Snippet: Primary antibodies were: anti-FGF21, AF3057, and anti-CD137, AF937 (R&D Systems, Minneapolis, MN), anti-P2RX5, sc-398682 (Santa Cruz Biotechnology, Dallas, TX), anti-beta-actin, sc-47778 (Santa Cruz, Dallas, TX), and anti-UCP1, ab10983, (abcam, Cambridge, MA).

Techniques: Marker

Journal: Nutrition & Metabolism

Article Title: Evaluation of markers of beige adipocytes in white adipose tissue of the mouse

doi: 10.1186/s12986-016-0081-2

Figure Lengend Snippet: List of primer/probe sets

Article Snippet: Primary antibodies were: anti-FGF21, AF3057, and anti-CD137, AF937 (R&D Systems, Minneapolis, MN), anti-P2RX5, sc-398682 (Santa Cruz Biotechnology, Dallas, TX), anti-beta-actin, sc-47778 (Santa Cruz, Dallas, TX), and anti-UCP1, ab10983, (abcam, Cambridge, MA).

Techniques:

Journal: Nutrition & Metabolism

Article Title: Evaluation of markers of beige adipocytes in white adipose tissue of the mouse

doi: 10.1186/s12986-016-0081-2

Figure Lengend Snippet: Markers of beige adipocytes in vitro . The transcripts for FGF21 a , P2RX5 b , PAT2 c , CITED1 d , CAR4 e , TBX1 f , CD137 g , and TMEM26 ( h ) were evaluated in white and beige adipocytes in culture treated with isoproterenol (+ Iso) or without (− Iso) for 6 hrs. before harvesting the cells. Transcripts were normalized to white – Iso. Values are means ± SEM ( n = 3). (*) different from white – Iso, (#) different from white + Iso, (+) different from beige + Iso, (@) different from beige – Iso, p ≤ 0.05, two way ANOVA followed by multiple comparisons using the Holm-Sidak method

Article Snippet: Primary antibodies were: anti-FGF21, AF3057, and anti-CD137, AF937 (R&D Systems, Minneapolis, MN), anti-P2RX5, sc-398682 (Santa Cruz Biotechnology, Dallas, TX), anti-beta-actin, sc-47778 (Santa Cruz, Dallas, TX), and anti-UCP1, ab10983, (abcam, Cambridge, MA).

Techniques: In Vitro

Journal: Nutrition & Metabolism

Article Title: Evaluation of markers of beige adipocytes in white adipose tissue of the mouse

doi: 10.1186/s12986-016-0081-2

Figure Lengend Snippet: Summary of beigeing criteria met by the thermoregulatory genes and putative beige markers when studied under in vitro and in vivo conditions

Article Snippet: Primary antibodies were: anti-FGF21, AF3057, and anti-CD137, AF937 (R&D Systems, Minneapolis, MN), anti-P2RX5, sc-398682 (Santa Cruz Biotechnology, Dallas, TX), anti-beta-actin, sc-47778 (Santa Cruz, Dallas, TX), and anti-UCP1, ab10983, (abcam, Cambridge, MA).

Techniques: In Vitro, In Vivo, Cell Culture

Journal: Nutrition & Metabolism

Article Title: Evaluation of markers of beige adipocytes in white adipose tissue of the mouse

doi: 10.1186/s12986-016-0081-2

Figure Lengend Snippet: Markers of beige adipocytes in inguinal adipose tissue. The transcripts for FGF21, P2RX5, PAT2, CITED1, CAR4, TBX1, CD137, and TMEM26 were evaluated in inguinal adipose tissue from mice maintained at room temperature (22 °C) or exposed to cold (5 °C) for one week in 3 separate experiments were the ages of the mice at the end of the experiment were 2 months and 10 days (Exp-1), 2 months and 23 days (Exp-2), and 4 months and 10 days (Exp-3). The levels of the transcripts at 5 °C were normalized to the transcript of the same gene at 22 °C. Values are means ± SEM ( n = 4–5). (*) different from 22 °C, p ≤ 0.05, two-tailed t-test

Article Snippet: Primary antibodies were: anti-FGF21, AF3057, and anti-CD137, AF937 (R&D Systems, Minneapolis, MN), anti-P2RX5, sc-398682 (Santa Cruz Biotechnology, Dallas, TX), anti-beta-actin, sc-47778 (Santa Cruz, Dallas, TX), and anti-UCP1, ab10983, (abcam, Cambridge, MA).

Techniques: Two Tailed Test

Journal: Nutrition & Metabolism

Article Title: Evaluation of markers of beige adipocytes in white adipose tissue of the mouse

doi: 10.1186/s12986-016-0081-2

Figure Lengend Snippet: Markers of beige adipocytes in the adipocyte and SVF derived from inguinal adipose tissue. The transcripts for FGF21 a , P2RX5 b , PAT2 c , CITED1 d , CAR4 e , TBX1 f , CD137 g , and TMEM26 ( h ) were evaluated in the adipocyte and SVF of 4 month old mice kept at room temperature (22 °C) or exposed to the cold (5 °C) for one week. Transcripts were normalized to 22 °C, SVF. Values are means ± SEM ( n = 3–5). (*) different from SVF 22 °C, (#) different from SVF 5 °C, (+) different from adipocytes 5 °C, (@) different from adipocytes 22 °C, p ≤ 0.05, two way ANOVA followed by multiple comparisons using the Holm-Sidak method

Article Snippet: Primary antibodies were: anti-FGF21, AF3057, and anti-CD137, AF937 (R&D Systems, Minneapolis, MN), anti-P2RX5, sc-398682 (Santa Cruz Biotechnology, Dallas, TX), anti-beta-actin, sc-47778 (Santa Cruz, Dallas, TX), and anti-UCP1, ab10983, (abcam, Cambridge, MA).

Techniques: Derivative Assay

Journal: Nutrition & Metabolism

Article Title: Evaluation of markers of beige adipocytes in white adipose tissue of the mouse

doi: 10.1186/s12986-016-0081-2

Figure Lengend Snippet: Beige markers in preadipocytes and SVF cells in vitro . The transcripts for CAR4, CITED1, PAT2, FGF21, P2RX5, CD137, TBX1, and TMEM26 were evaluated in SVF cells and beige adipocytes in culture. Transcripts were normalized to SVF cells. Values are means ± SEM ( n = 3). (*) different from SVF, p ≤ 0.05, two-tailed t-test

Article Snippet: Primary antibodies were: anti-FGF21, AF3057, and anti-CD137, AF937 (R&D Systems, Minneapolis, MN), anti-P2RX5, sc-398682 (Santa Cruz Biotechnology, Dallas, TX), anti-beta-actin, sc-47778 (Santa Cruz, Dallas, TX), and anti-UCP1, ab10983, (abcam, Cambridge, MA).

Techniques: In Vitro, Two Tailed Test