4 1bb fusion protein treatment100  (Sino Biological)


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    Human respiratory syncytial virus Fusion glycoprotein RSV F Gene Lentiviral ORF cDNA expression plasmid C GFPSpark tag
    Description:
    Full length Clone DNA of Human RSV subtype A strain Long Fusion glycoprotein RSV F
    Catalog Number:
    VG40039-ACGLN
    Price:
    525.0
    Category:
    cDNA Clone
    Size:
    1Unit
    Product Aliases:
    F cDNA ORF Clone RSV, HRSVgp08 cDNA ORF Clone RSV
    Molecule Name:
    RSV-F,F,RSV Fusion,
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    Structured Review

    Sino Biological 4 1bb fusion protein treatment100
    Human respiratory syncytial virus Fusion glycoprotein RSV F Gene Lentiviral ORF cDNA expression plasmid C GFPSpark tag
    Full length Clone DNA of Human RSV subtype A strain Long Fusion glycoprotein RSV F
    https://www.bioz.com/result/4 1bb fusion protein treatment100/product/Sino Biological
    Average 93 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    4 1bb fusion protein treatment100 - by Bioz Stars, 2021-08
    93/100 stars

    Images

    1) Product Images from "4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice"

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

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.01848

    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.
    Figure Legend 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.

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

    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.
    Figure Legend 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.

    Techniques Used: Affinity Magnetic Separation, 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).
    Figure Legend 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).

    Techniques Used: 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).
    Figure Legend 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).

    Techniques Used: Expressing, Western Blot, Isolation

    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).
    Figure Legend 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).

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

    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).
    Figure Legend 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).

    Techniques Used: Mouse Assay, Inhibition, Injection, Western Blot, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining, Expressing

    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).
    Figure Legend 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).

    Techniques Used: Expressing, Mouse Assay, Affinity Magnetic Separation

    2) Product Images from "Active immunoprophylaxis with a synthetic DNA-encoded monoclonal anti-respiratory syncytial virus scFv-Fc fusion protein confers protection against infection and durable activity"

    Article Title: Active immunoprophylaxis with a synthetic DNA-encoded monoclonal anti-respiratory syncytial virus scFv-Fc fusion protein confers protection against infection and durable activity

    Journal: Human Vaccines & Immunotherapeutics

    doi: 10.1080/21645515.2020.1748979

    Schematic of RSV-F dMAb in-vivo delivery platform : (a) In vivo EP facilitates the delivery of the dMAb to the muscle tissue, myocytes express (b) and secret the encoded mAb (c). mAb is distributed systemically. (b) Example of expression of dMAb in myocytes 3 days after delivery of 50 µg dMAb or pVax (empty expression vector control) to mouse TA (DAPI – blue, dMAb – green). (c) Serum concentration of RSV-F dMAb and Palivizumab after delivery of 200 µg RSV-F dMAb plasmid or 15 mg/kg (clinical dose) Palivizumab measured by ELISA (± SEM, n = 6) dotted line indicates the level of protection (20 µg/ml) for Palivizumab.
    Figure Legend Snippet: Schematic of RSV-F dMAb in-vivo delivery platform : (a) In vivo EP facilitates the delivery of the dMAb to the muscle tissue, myocytes express (b) and secret the encoded mAb (c). mAb is distributed systemically. (b) Example of expression of dMAb in myocytes 3 days after delivery of 50 µg dMAb or pVax (empty expression vector control) to mouse TA (DAPI – blue, dMAb – green). (c) Serum concentration of RSV-F dMAb and Palivizumab after delivery of 200 µg RSV-F dMAb plasmid or 15 mg/kg (clinical dose) Palivizumab measured by ELISA (± SEM, n = 6) dotted line indicates the level of protection (20 µg/ml) for Palivizumab.

    Techniques Used: In Vivo, Expressing, Plasmid Preparation, Concentration Assay, Enzyme-linked Immunosorbent Assay

    Design and in-vitro testing of scFv-Fc RSV-F dMAb construct : (a) Illustration of human IgG1 and scFv-Fc formats. The complete hinge and Fc portions of the molecule are retained in the scFv-Fc format. Yellow line connecting VH and VL of the scFv-Fc molecule represents the (G4 S) 3 linker (b) Plasmid map of RSV-F dMAb (bGH PolyA – bovine growth hormone polyadenylation signal, KanR – kanamycin resistance gene, pUC ori – pUC origin of replication, hCMV promotor – human cytomegalovirus promotor) (c) In vitro expression of RSV-F dMAb. Immuno-fluorescence staining was performed on HEK293 T cells 3 days after in vitro transfection (DAPI – blue, RSV-F dMAb – red) (d) Western-blot analysis of cell-culture supernatant of RSV-F dMAb in vitro transfected HEK293 T cells harvested on day 3 post-transfection (lane 1: IgG-RSV dMAb, lane 2: scFv-Fc RSV-F dMAb).
    Figure Legend Snippet: Design and in-vitro testing of scFv-Fc RSV-F dMAb construct : (a) Illustration of human IgG1 and scFv-Fc formats. The complete hinge and Fc portions of the molecule are retained in the scFv-Fc format. Yellow line connecting VH and VL of the scFv-Fc molecule represents the (G4 S) 3 linker (b) Plasmid map of RSV-F dMAb (bGH PolyA – bovine growth hormone polyadenylation signal, KanR – kanamycin resistance gene, pUC ori – pUC origin of replication, hCMV promotor – human cytomegalovirus promotor) (c) In vitro expression of RSV-F dMAb. Immuno-fluorescence staining was performed on HEK293 T cells 3 days after in vitro transfection (DAPI – blue, RSV-F dMAb – red) (d) Western-blot analysis of cell-culture supernatant of RSV-F dMAb in vitro transfected HEK293 T cells harvested on day 3 post-transfection (lane 1: IgG-RSV dMAb, lane 2: scFv-Fc RSV-F dMAb).

    Techniques Used: In Vitro, Construct, Plasmid Preparation, Expressing, Fluorescence, Staining, Transfection, Western Blot, Cell Culture

    Characterization of RSV-F dMAb in cotton rats: (a) Local expression of dMAb demonstrated by Immuno-fluorescence staining of RSV-F dMAb in cotton rat TA muscle tissue 7 days after delivery of 400 µg RSV-F dMAb pDNA to the tissue site (DAPI – blue; RSV-F dMAb – green; sectioned perpendicular to myocytes) (b) Levels of RSV-F dMAb (ng/ml) was measured for 39 days after IM administration of 800 µg dMAb-plasmid (±SEM, n = 5). (c) Virus neutralization function of in-vivo expressed RSV-F dMAb. Serum samples were harvested and tested 7 days after delivery of 2.4 mg scFv-Fc dMAb-pDNA: neutralizing titer (±SEM, dotted line indicates LOD at serum-dilution of 1/20). (d) Concentration of RSV-F dMAb in BAL samples from treated cotton rats.
    Figure Legend Snippet: Characterization of RSV-F dMAb in cotton rats: (a) Local expression of dMAb demonstrated by Immuno-fluorescence staining of RSV-F dMAb in cotton rat TA muscle tissue 7 days after delivery of 400 µg RSV-F dMAb pDNA to the tissue site (DAPI – blue; RSV-F dMAb – green; sectioned perpendicular to myocytes) (b) Levels of RSV-F dMAb (ng/ml) was measured for 39 days after IM administration of 800 µg dMAb-plasmid (±SEM, n = 5). (c) Virus neutralization function of in-vivo expressed RSV-F dMAb. Serum samples were harvested and tested 7 days after delivery of 2.4 mg scFv-Fc dMAb-pDNA: neutralizing titer (±SEM, dotted line indicates LOD at serum-dilution of 1/20). (d) Concentration of RSV-F dMAb in BAL samples from treated cotton rats.

    Techniques Used: Expressing, Fluorescence, Staining, Plasmid Preparation, Neutralization, In Vivo, Concentration Assay

    In-vivo expression of RSV-F dMAb in BALB/c mice : 200 µg RSV-F dMAb plasmid was administered IM in BALB/c mice (a-d). Serum samples were taken from treated mice before treatment (day 0) and 7 days after treatment to measure (a) Serum expression of RSV-F dMAb. scFv-Fc construct (± SEM, n = 5–8) (b) RSV-F antigen binding of dMAb. RSV-F binding signal of serum samples from treated (squares) and naïve (triangles) mice (± SEM, n = 4) (c) In vitro neutralization of RSV-A virus. Neutralization titer of serum samples from treated (squares) or naïve (triangles) mice (log2 serum dilution of 60% reduction of plaque-formation; ±SEM, n = 3–11, dotted line indicates LOD at serum-dilution of 1/20) (d) Concentration of scFv-Fc RSV-F dMAb in BAL samples 7 days after RSV-dMAb pDNA delivery in treated (squares) or naïve (triangles) mice (mol scFv-Fc dMAb per gram total protein in lavage sample, n(naïve) = 2, n(RSV-F dMAb) = 3).
    Figure Legend Snippet: In-vivo expression of RSV-F dMAb in BALB/c mice : 200 µg RSV-F dMAb plasmid was administered IM in BALB/c mice (a-d). Serum samples were taken from treated mice before treatment (day 0) and 7 days after treatment to measure (a) Serum expression of RSV-F dMAb. scFv-Fc construct (± SEM, n = 5–8) (b) RSV-F antigen binding of dMAb. RSV-F binding signal of serum samples from treated (squares) and naïve (triangles) mice (± SEM, n = 4) (c) In vitro neutralization of RSV-A virus. Neutralization titer of serum samples from treated (squares) or naïve (triangles) mice (log2 serum dilution of 60% reduction of plaque-formation; ±SEM, n = 3–11, dotted line indicates LOD at serum-dilution of 1/20) (d) Concentration of scFv-Fc RSV-F dMAb in BAL samples 7 days after RSV-dMAb pDNA delivery in treated (squares) or naïve (triangles) mice (mol scFv-Fc dMAb per gram total protein in lavage sample, n(naïve) = 2, n(RSV-F dMAb) = 3).

    Techniques Used: In Vivo, Expressing, Mouse Assay, Plasmid Preparation, Construct, Binding Assay, In Vitro, Neutralization, Concentration Assay

    RSV-F dMAb confers protection against LRD after RSV/A challenge of cotton rats: (a) Schematic of cotton rat challenge study: Animals were treated with 2.4 mg RSV-dMAb 7 days before challenge or with an IM injection of 15 mg/kg Palivizumab 1 day before challenge (b) Viral load of cotton rat lung tissue (pfu/g) harvested 5 days after intra-nasal live virus challenge with RSV/A/long (± SEM n = 4–5) (i). RSV Nonstructural protein-1 (NS-1) mRNA levels (log2 and normalized to beta-Actin) of cotton rat lung tissue 5 days after intra-nasal live virus challenge with RSV/A/long (± SEM, n = 4–5, Mann–Whitney non-parametric t-test: p (untreated vs RSV-dMAb) = 0.0159; p (untreated vs Palivizumab) = 0.0079) (ii). (c) Serum levels of Palivizumab and scFv-Fc RSV-F dMAb in cotton rats at the day of challenge (day 0) and 5 days after challenge (± SEM, n = 4–5).
    Figure Legend Snippet: RSV-F dMAb confers protection against LRD after RSV/A challenge of cotton rats: (a) Schematic of cotton rat challenge study: Animals were treated with 2.4 mg RSV-dMAb 7 days before challenge or with an IM injection of 15 mg/kg Palivizumab 1 day before challenge (b) Viral load of cotton rat lung tissue (pfu/g) harvested 5 days after intra-nasal live virus challenge with RSV/A/long (± SEM n = 4–5) (i). RSV Nonstructural protein-1 (NS-1) mRNA levels (log2 and normalized to beta-Actin) of cotton rat lung tissue 5 days after intra-nasal live virus challenge with RSV/A/long (± SEM, n = 4–5, Mann–Whitney non-parametric t-test: p (untreated vs RSV-dMAb) = 0.0159; p (untreated vs Palivizumab) = 0.0079) (ii). (c) Serum levels of Palivizumab and scFv-Fc RSV-F dMAb in cotton rats at the day of challenge (day 0) and 5 days after challenge (± SEM, n = 4–5).

    Techniques Used: Injection, MANN-WHITNEY

    3) Product Images from "A Novel Inactivated Intranasal Respiratory Syncytial Virus Vaccine Promotes Viral Clearance without Th2 Associated Vaccine-Enhanced Disease"

    Article Title: A Novel Inactivated Intranasal Respiratory Syncytial Virus Vaccine Promotes Viral Clearance without Th2 Associated Vaccine-Enhanced Disease

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0021823

    Intranasal vaccination of mice with NE-RSV results in RSV-specific antibody responses. Mice were immunized with NE-RSV containing 10 5 virus particles at Day 0 and Day 28. In (A), the levels of RSV specific antibodies (IgG+IgM) in serum were determined at weeks 2, 4, 6, 8, and 10 via ELISA using purified RSV protein. In (B), serum samples were serially diluted to obtain endpoints titers. In (C), RSV-specific IgA, IgG2a, IgG1, and IgE were assessed at week 8 via isotype-specific ELISA. In (D), IgG2a and IgG1 antibodies specific for RSV F and G were assessed at week 8 via isotype-specific ELISA using purified RSV F and G glycoproteins. In (E), RSV-specific IgA responses in bronchoalveolar lavage samples (BAL) were assessed using an isotype-specific ELISA. Each time point represents the mean of a minimum of 5 samples +/− SEM, and the experiment was repeated with similar results. * = P
    Figure Legend Snippet: Intranasal vaccination of mice with NE-RSV results in RSV-specific antibody responses. Mice were immunized with NE-RSV containing 10 5 virus particles at Day 0 and Day 28. In (A), the levels of RSV specific antibodies (IgG+IgM) in serum were determined at weeks 2, 4, 6, 8, and 10 via ELISA using purified RSV protein. In (B), serum samples were serially diluted to obtain endpoints titers. In (C), RSV-specific IgA, IgG2a, IgG1, and IgE were assessed at week 8 via isotype-specific ELISA. In (D), IgG2a and IgG1 antibodies specific for RSV F and G were assessed at week 8 via isotype-specific ELISA using purified RSV F and G glycoproteins. In (E), RSV-specific IgA responses in bronchoalveolar lavage samples (BAL) were assessed using an isotype-specific ELISA. Each time point represents the mean of a minimum of 5 samples +/− SEM, and the experiment was repeated with similar results. * = P

    Techniques Used: Mouse Assay, Enzyme-linked Immunosorbent Assay, Purification

    4) Product Images from "Age-Specific Profiles of Antibody Responses against Respiratory Syncytial Virus Infection"

    Article Title: Age-Specific Profiles of Antibody Responses against Respiratory Syncytial Virus Infection

    Journal: EBioMedicine

    doi: 10.1016/j.ebiom.2017.01.014

    Evaluation of serum IgG to recognize antigenic sites I, II, and IV in F protein. Sera were subjected to competitive ELISA by using monoclonal antibody specific to antigenic sites I, II, or IV. The amounts of serum IgG specific for each antigenic epitopes are represented by IC50 resulting in 50% reduction of biding RSV F with competitive monoclonal antibody. The limitation of the IC50 detection was defined as 5. The concentration of serum IgG specific to antigenic site II was calculated based on the result of palivizumab on competitive ELISA. The mean ± 95% CI was estimated by the linear model, and were shown in the graphs. IC50 of antigenic site I-specific IgG in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 26; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 32; adults, n = 23. IC50 of antigenic site I-specific IgG in convalescent phase: 0–3 mo, n = 24; 4–6 mo, n = 27; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 32; adults, n = 23. GMRs C/A of antigenic site I-specific IgG: 0–3 mo, n = 21; 4–6 mo, n = 25; 7–12 mo, n = 26; 13–18 mo, n = 34; 19–36 mo, n = 31; adults, n = 23. IC50 of antigenic site II-specific IgG in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 28; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 33; adults, n = 23. IC50 of antigenic site II-specific IgG in convalescent phase: 0–3 mo, n = 27; 4–6 mo, n = 29; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 33; adults, n = 23. GMRs C/A of antigenic site II-specific IgG: 0–3 mo, n = 24; 4–6 mo, n = 27; 7–12 mo, n = 26; 13–18 mo, n = 34; 19–36 mo, n = 32; adults, n = 23. IC50 of antigenic site IV-specific IgG in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 28; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 31; adults, n = 23. IC50 of antigenic site IV-specific IgG in convalescent phase: 0–3 mo, n = 27; 4–6 mo, n = 29; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 33; adults, n = 23. GMRs C/A of antigenic site IV-specific IgG: 0–3 mo, n = 24; 4–6 mo, n = 27; 7–12 mo, n = 26; 13–18 mo, n = 34; 19–36 mo, n = 31; adults, n = 23. * p
    Figure Legend Snippet: Evaluation of serum IgG to recognize antigenic sites I, II, and IV in F protein. Sera were subjected to competitive ELISA by using monoclonal antibody specific to antigenic sites I, II, or IV. The amounts of serum IgG specific for each antigenic epitopes are represented by IC50 resulting in 50% reduction of biding RSV F with competitive monoclonal antibody. The limitation of the IC50 detection was defined as 5. The concentration of serum IgG specific to antigenic site II was calculated based on the result of palivizumab on competitive ELISA. The mean ± 95% CI was estimated by the linear model, and were shown in the graphs. IC50 of antigenic site I-specific IgG in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 26; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 32; adults, n = 23. IC50 of antigenic site I-specific IgG in convalescent phase: 0–3 mo, n = 24; 4–6 mo, n = 27; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 32; adults, n = 23. GMRs C/A of antigenic site I-specific IgG: 0–3 mo, n = 21; 4–6 mo, n = 25; 7–12 mo, n = 26; 13–18 mo, n = 34; 19–36 mo, n = 31; adults, n = 23. IC50 of antigenic site II-specific IgG in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 28; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 33; adults, n = 23. IC50 of antigenic site II-specific IgG in convalescent phase: 0–3 mo, n = 27; 4–6 mo, n = 29; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 33; adults, n = 23. GMRs C/A of antigenic site II-specific IgG: 0–3 mo, n = 24; 4–6 mo, n = 27; 7–12 mo, n = 26; 13–18 mo, n = 34; 19–36 mo, n = 32; adults, n = 23. IC50 of antigenic site IV-specific IgG in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 28; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 31; adults, n = 23. IC50 of antigenic site IV-specific IgG in convalescent phase: 0–3 mo, n = 27; 4–6 mo, n = 29; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 33; adults, n = 23. GMRs C/A of antigenic site IV-specific IgG: 0–3 mo, n = 24; 4–6 mo, n = 27; 7–12 mo, n = 26; 13–18 mo, n = 34; 19–36 mo, n = 31; adults, n = 23. * p

    Techniques Used: Competitive ELISA, Concentration Assay

    Serum antibody response to RSV F protein. The sera were subjected to ELISA coating RSV F protein. Series of detection antibody labeled with HRP were used to acquire the profile of IgG subclasses specific to RSV F protein. A left panel represents concentration of total IgG, IgG1, IgG2 and IgG3 for RSV F in acute phase or convalescent phase, and a right panel does the GMRs C/A of F IgGtotal, IgG1, IgG2, and IgG3. The mean ± 95% CI was estimated by the linear model, and were shown in the graphs. Concentration of antibody in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 28; 7–12 mo, n = 29; 13–18 mo, n = 34; 19–36 mo, n = 35; adults, n = 23. Concentration of antibody in convalescent phase: 0–3 mo, n = 28; 4–6 mo, n = 30; 7–12 mo, n = 33; 13–18 mo, n = 35; 19–36 mo, n = 34; adults, n = 23. GMRs C/A: 0–3 mo, n = 23; 4–6 mo, n = 28; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 34; adults, n = 23. * p
    Figure Legend Snippet: Serum antibody response to RSV F protein. The sera were subjected to ELISA coating RSV F protein. Series of detection antibody labeled with HRP were used to acquire the profile of IgG subclasses specific to RSV F protein. A left panel represents concentration of total IgG, IgG1, IgG2 and IgG3 for RSV F in acute phase or convalescent phase, and a right panel does the GMRs C/A of F IgGtotal, IgG1, IgG2, and IgG3. The mean ± 95% CI was estimated by the linear model, and were shown in the graphs. Concentration of antibody in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 28; 7–12 mo, n = 29; 13–18 mo, n = 34; 19–36 mo, n = 35; adults, n = 23. Concentration of antibody in convalescent phase: 0–3 mo, n = 28; 4–6 mo, n = 30; 7–12 mo, n = 33; 13–18 mo, n = 35; 19–36 mo, n = 34; adults, n = 23. GMRs C/A: 0–3 mo, n = 23; 4–6 mo, n = 28; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 34; adults, n = 23. * p

    Techniques Used: Enzyme-linked Immunosorbent Assay, Labeling, Concentration Assay

    Antibody affinity to RSV F protein. Sera were subjected to a surface plasmon resonance (SPR) assay. Apparent dissociation rate constants ( k d ) were calculated and shown in different age groups. The mean ± 95% CI was estimated by linear model, and were shown in the graphs. k d in acute phase: 0–3 mo, n = 30; 4–6 mo, n = 29; 7–12 mo, n = 30; 13–18 mo, n = 30; 19–36 mo, n = 32; adults, n = 22. k d in convalescent phase: 0–3 mo, n = 29; 4–6 mo, n = 30; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 33; adults, n = 23. GMRs C/A of F IgG k d : 0–3 mo, n = 25; 4–6 mo, n = 29; 7–12 mo, n = 28; 13–18 mo, n = 30; 19–36 mo, n = 31; adults, n = 22. * p
    Figure Legend Snippet: Antibody affinity to RSV F protein. Sera were subjected to a surface plasmon resonance (SPR) assay. Apparent dissociation rate constants ( k d ) were calculated and shown in different age groups. The mean ± 95% CI was estimated by linear model, and were shown in the graphs. k d in acute phase: 0–3 mo, n = 30; 4–6 mo, n = 29; 7–12 mo, n = 30; 13–18 mo, n = 30; 19–36 mo, n = 32; adults, n = 22. k d in convalescent phase: 0–3 mo, n = 29; 4–6 mo, n = 30; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 33; adults, n = 23. GMRs C/A of F IgG k d : 0–3 mo, n = 25; 4–6 mo, n = 29; 7–12 mo, n = 28; 13–18 mo, n = 30; 19–36 mo, n = 31; adults, n = 22. * p

    Techniques Used: SPR Assay

    5) Product Images from "Virus-like particle vaccines containing F or F and G proteins confer protection against respiratory syncytial virus without pulmonary inflammation in cotton rats"

    Article Title: Virus-like particle vaccines containing F or F and G proteins confer protection against respiratory syncytial virus without pulmonary inflammation in cotton rats

    Journal: Human Vaccines & Immunotherapeutics

    doi: 10.1080/21645515.2016.1272743

    FG VLP is effective in inducing RSV (F)protein-specific antibodies in cotton rats. (A) Prime IgG antibodies specific for RSV F protein. (B) Boost IgG antibodies specific for RSV F protein. (C) Prime IgG antibodies specific for RSV. (D) Boost IgG antibodies specific for RSV. (E) Prime IgG antibodies specific for RSV G protein. (F) Boost IgG antibodies specific for RSV G protein. Cotton rats (n = 5 per group) were immunized i.m. with F VLP, G VLP, mixed F VLP and G VLP (FG VLP), FI-RSV (FI-RSV), intranasally incoculated with live RSV (0.3 × 10 6 PFU). and PBS (Naïve) on days 0 (prime) and 28 (boost). Serum samples were collected at 3 weeks after prime or boost immunization and RSV-specific antibody levels were measured by ELISA. Results are presented as mean ± SEM and statistical significance was performed by one-way ANOVA with Tukey's multiple comparisons post-test in Graph Pad Prism. *** p
    Figure Legend Snippet: FG VLP is effective in inducing RSV (F)protein-specific antibodies in cotton rats. (A) Prime IgG antibodies specific for RSV F protein. (B) Boost IgG antibodies specific for RSV F protein. (C) Prime IgG antibodies specific for RSV. (D) Boost IgG antibodies specific for RSV. (E) Prime IgG antibodies specific for RSV G protein. (F) Boost IgG antibodies specific for RSV G protein. Cotton rats (n = 5 per group) were immunized i.m. with F VLP, G VLP, mixed F VLP and G VLP (FG VLP), FI-RSV (FI-RSV), intranasally incoculated with live RSV (0.3 × 10 6 PFU). and PBS (Naïve) on days 0 (prime) and 28 (boost). Serum samples were collected at 3 weeks after prime or boost immunization and RSV-specific antibody levels were measured by ELISA. Results are presented as mean ± SEM and statistical significance was performed by one-way ANOVA with Tukey's multiple comparisons post-test in Graph Pad Prism. *** p

    Techniques Used: Enzyme-linked Immunosorbent Assay

    FG VLP or (F)VLP vaccination is effective in inducing RSV F-specific antibodies and antibody secreting cells. (A) BM cells secreting F specific IgG antibodies. (B) MLN cells secreting F specific IgG antibodies. (C) Spleen cells secreting F specific IgG antibodies. Cells from BM, spleens, and MLN were incubated in the culture plates coated with RSV F protein (400 ng/ml) for 2 d. Secreted antibodies were detected by ELISA analysis. RSV F-protein specific l IgG antibody responses in BALF (D) and lung extracts (E) were determined by ELISA. Results are presented as mean ± SEM and statistical significance was performed by one-way ANOVA with Tukey's multiple comparisons post-test in Graph Pad Prism; *** p
    Figure Legend Snippet: FG VLP or (F)VLP vaccination is effective in inducing RSV F-specific antibodies and antibody secreting cells. (A) BM cells secreting F specific IgG antibodies. (B) MLN cells secreting F specific IgG antibodies. (C) Spleen cells secreting F specific IgG antibodies. Cells from BM, spleens, and MLN were incubated in the culture plates coated with RSV F protein (400 ng/ml) for 2 d. Secreted antibodies were detected by ELISA analysis. RSV F-protein specific l IgG antibody responses in BALF (D) and lung extracts (E) were determined by ELISA. Results are presented as mean ± SEM and statistical significance was performed by one-way ANOVA with Tukey's multiple comparisons post-test in Graph Pad Prism; *** p

    Techniques Used: Incubation, Enzyme-linked Immunosorbent Assay

    6) Product Images from "SPLUNC1 Is a Significant Marker in Pleural Effusion from Lung Cancer Compared to Tuberculosis"

    Article Title: SPLUNC1 Is a Significant Marker in Pleural Effusion from Lung Cancer Compared to Tuberculosis

    Journal: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy

    doi: 10.1089/mab.2014.0073

    Specificity of prepared MAbs against SPLUNC1. ( A ) Specific identification by ELISA. SPLUNC1-hFc, mE3-hFc, and human IgG1-Fc were separately coated, and MAbs from different clones were added. 1, clone 9; 2, clone 130; 3, clone 160; 4, clone 104; 5, positive
    Figure Legend Snippet: Specificity of prepared MAbs against SPLUNC1. ( A ) Specific identification by ELISA. SPLUNC1-hFc, mE3-hFc, and human IgG1-Fc were separately coated, and MAbs from different clones were added. 1, clone 9; 2, clone 130; 3, clone 160; 4, clone 104; 5, positive

    Techniques Used: Enzyme-linked Immunosorbent Assay, Clone Assay

    Related Articles

    Binding Assay:

    Article Title: Active immunoprophylaxis with a synthetic DNA-encoded monoclonal anti-respiratory syncytial virus scFv-Fc fusion protein confers protection against infection and durable activity
    Article Snippet: .. Antigen binding ELISAAssay plates were coated with 1 µg/well of human RSV F glycoprotein (Sino Biological, Wayne, PA) diluted in 1x DPBS (ThermoFisher) overnight at 4°C. ..

    Produced:

    Article Title: SPLUNC1 Is a Significant Marker in Pleural Effusion from Lung Cancer Compared to Tuberculosis
    Article Snippet: .. A control fusion protein mE3-hFc, which was mouse CD137 cysteine-rich domains with human IgG-Fc, was produced by our laboratory,( ) and another control of human IgG1-Fc protein was purchased from Sino Biological (Beijing, China). ..

    Purification:

    Article Title: A Novel Inactivated Intranasal Respiratory Syncytial Virus Vaccine Promotes Viral Clearance without Th2 Associated Vaccine-Enhanced Disease
    Article Snippet: .. RSV F and G specific ELISAs were performed similarly, except that plates were coated with purified recombinant RSV F or RSV G glycoproteins (1.3 ug/ml) expressed in Baculovirus (Sino Biological). ..

    Recombinant:

    Article Title: A Novel Inactivated Intranasal Respiratory Syncytial Virus Vaccine Promotes Viral Clearance without Th2 Associated Vaccine-Enhanced Disease
    Article Snippet: .. RSV F and G specific ELISAs were performed similarly, except that plates were coated with purified recombinant RSV F or RSV G glycoproteins (1.3 ug/ml) expressed in Baculovirus (Sino Biological). ..

    Enzyme-linked Immunosorbent Assay:

    Article Title: Age-Specific Profiles of Antibody Responses against Respiratory Syncytial Virus Infection
    Article Snippet: .. 2.4 Enzyme-linked ImmunoSorbent Assay (ELISA) of RSV F and G Protein The 96-well plates were coated with 0.5 μg/mL of F or G protein (SinoBiological Inc.) and serially diluted human IgG protein (Nordic Immunology, Human Standard Serum NOR-01) at 4 °C for 12 h to 18 h. After washing with PBS (-) containing 0.05% Tween 20 (PBST), the plates were treated with the blocking solution (1% BSA in PBST). ..

    Article Title: Virus-like particle vaccines containing F or F and G proteins confer protection against respiratory syncytial virus without pulmonary inflammation in cotton rats
    Article Snippet: .. RSV F specific antibodies and G specific antibodies (IgG) in primed and boosted sera were determined by ELISA using RSV F (BEI, NIAID, NIH) or G protein (Sino Biological, PA) as a coating antigen. ..

    Blocking Assay:

    Article Title: Age-Specific Profiles of Antibody Responses against Respiratory Syncytial Virus Infection
    Article Snippet: .. 2.4 Enzyme-linked ImmunoSorbent Assay (ELISA) of RSV F and G Protein The 96-well plates were coated with 0.5 μg/mL of F or G protein (SinoBiological Inc.) and serially diluted human IgG protein (Nordic Immunology, Human Standard Serum NOR-01) at 4 °C for 12 h to 18 h. After washing with PBS (-) containing 0.05% Tween 20 (PBST), the plates were treated with the blocking solution (1% BSA in PBST). ..

    Amplification:

    Article Title: Gene delivery of hypoxia-inducible VEGF targeting collagen effectively improves cardiac function after myocardial infarction
    Article Snippet: .. The ORF of human VEGF-A-Flag fusion fragment without the termination codon was amplified by fusion PCR primer pairs (vegf-F/vegf-R) from the DNA template of pCMV3-VEGFA-Flag plasmid (Sino Biological Inc. Beijing, China, #HG11066-CF), which consists of the human VEGF-A (GenBank No. NM_001171626.1) coding sequence and a Flag tag. ..

    Polymerase Chain Reaction:

    Article Title: Gene delivery of hypoxia-inducible VEGF targeting collagen effectively improves cardiac function after myocardial infarction
    Article Snippet: .. The ORF of human VEGF-A-Flag fusion fragment without the termination codon was amplified by fusion PCR primer pairs (vegf-F/vegf-R) from the DNA template of pCMV3-VEGFA-Flag plasmid (Sino Biological Inc. Beijing, China, #HG11066-CF), which consists of the human VEGF-A (GenBank No. NM_001171626.1) coding sequence and a Flag tag. ..

    Plasmid Preparation:

    Article Title: Gene delivery of hypoxia-inducible VEGF targeting collagen effectively improves cardiac function after myocardial infarction
    Article Snippet: .. The ORF of human VEGF-A-Flag fusion fragment without the termination codon was amplified by fusion PCR primer pairs (vegf-F/vegf-R) from the DNA template of pCMV3-VEGFA-Flag plasmid (Sino Biological Inc. Beijing, China, #HG11066-CF), which consists of the human VEGF-A (GenBank No. NM_001171626.1) coding sequence and a Flag tag. ..

    Sequencing:

    Article Title: Gene delivery of hypoxia-inducible VEGF targeting collagen effectively improves cardiac function after myocardial infarction
    Article Snippet: .. The ORF of human VEGF-A-Flag fusion fragment without the termination codon was amplified by fusion PCR primer pairs (vegf-F/vegf-R) from the DNA template of pCMV3-VEGFA-Flag plasmid (Sino Biological Inc. Beijing, China, #HG11066-CF), which consists of the human VEGF-A (GenBank No. NM_001171626.1) coding sequence and a Flag tag. ..

    FLAG-tag:

    Article Title: Gene delivery of hypoxia-inducible VEGF targeting collagen effectively improves cardiac function after myocardial infarction
    Article Snippet: .. The ORF of human VEGF-A-Flag fusion fragment without the termination codon was amplified by fusion PCR primer pairs (vegf-F/vegf-R) from the DNA template of pCMV3-VEGFA-Flag plasmid (Sino Biological Inc. Beijing, China, #HG11066-CF), which consists of the human VEGF-A (GenBank No. NM_001171626.1) coding sequence and a Flag tag. ..

    Injection:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: .. For 4-1BB Fusion Protein Treatment100 or 50 µg of 4-1BB fusion protein (4-1BBIg; cat: 50811-M02H, Sino Biological Inc., Beijing, China) per mouse or 100 µg isotype IgG (human IgG1; Sino Biological Inc.) were injected intraperitoneally into mice, on days 1 and 4 after CS administration (3–4 mice per group). ..

    Mouse Assay:

    Article Title: 4-1BB Signaling Promotes Alveolar Macrophages-Mediated Pro-Fibrotic Responses and Crystalline Silica-Induced Pulmonary Fibrosis in Mice
    Article Snippet: .. For 4-1BB Fusion Protein Treatment100 or 50 µg of 4-1BB fusion protein (4-1BBIg; cat: 50811-M02H, Sino Biological Inc., Beijing, China) per mouse or 100 µg isotype IgG (human IgG1; Sino Biological Inc.) were injected intraperitoneally into mice, on days 1 and 4 after CS administration (3–4 mice per group). ..

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    Sino Biological 4 1bb fusion protein treatment100
    Blockade of <t>4-1BB</t> 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.
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    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: For 4-1BB Fusion Protein Treatment100 or 50 µg of 4-1BB fusion protein (4-1BBIg; cat: 50811-M02H, Sino Biological Inc., Beijing, China) per mouse or 100 µg isotype IgG (human IgG1; Sino Biological Inc.) were injected intraperitoneally into mice, on days 1 and 4 after CS administration (3–4 mice per group).

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

    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: For 4-1BB Fusion Protein Treatment100 or 50 µg of 4-1BB fusion protein (4-1BBIg; cat: 50811-M02H, Sino Biological Inc., Beijing, China) per mouse or 100 µg isotype IgG (human IgG1; Sino Biological Inc.) were injected intraperitoneally into mice, on days 1 and 4 after CS administration (3–4 mice per group).

    Techniques: Affinity Magnetic Separation, 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: For 4-1BB Fusion Protein Treatment100 or 50 µg of 4-1BB fusion protein (4-1BBIg; cat: 50811-M02H, Sino Biological Inc., Beijing, China) per mouse or 100 µg isotype IgG (human IgG1; Sino Biological Inc.) were injected intraperitoneally into mice, on days 1 and 4 after CS administration (3–4 mice per group).

    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: For 4-1BB Fusion Protein Treatment100 or 50 µg of 4-1BB fusion protein (4-1BBIg; cat: 50811-M02H, Sino Biological Inc., Beijing, China) per mouse or 100 µg isotype IgG (human IgG1; Sino Biological Inc.) were injected intraperitoneally into mice, on days 1 and 4 after CS administration (3–4 mice per group).

    Techniques: Expressing, Western Blot, Isolation

    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: For 4-1BB Fusion Protein Treatment100 or 50 µg of 4-1BB fusion protein (4-1BBIg; cat: 50811-M02H, Sino Biological Inc., Beijing, China) per mouse or 100 µg isotype IgG (human IgG1; Sino Biological Inc.) were injected intraperitoneally into mice, on days 1 and 4 after CS administration (3–4 mice per group).

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

    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: For 4-1BB Fusion Protein Treatment100 or 50 µg of 4-1BB fusion protein (4-1BBIg; cat: 50811-M02H, Sino Biological Inc., Beijing, China) per mouse or 100 µg isotype IgG (human IgG1; Sino Biological Inc.) were injected intraperitoneally into mice, on days 1 and 4 after CS administration (3–4 mice per group).

    Techniques: Mouse Assay, Inhibition, Injection, Western Blot, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Staining, Expressing

    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: For 4-1BB Fusion Protein Treatment100 or 50 µg of 4-1BB fusion protein (4-1BBIg; cat: 50811-M02H, Sino Biological Inc., Beijing, China) per mouse or 100 µg isotype IgG (human IgG1; Sino Biological Inc.) were injected intraperitoneally into mice, on days 1 and 4 after CS administration (3–4 mice per group).

    Techniques: Expressing, Mouse Assay, Affinity Magnetic Separation

    Schematic of RSV-F dMAb in-vivo delivery platform : (a) In vivo EP facilitates the delivery of the dMAb to the muscle tissue, myocytes express (b) and secret the encoded mAb (c). mAb is distributed systemically. (b) Example of expression of dMAb in myocytes 3 days after delivery of 50 µg dMAb or pVax (empty expression vector control) to mouse TA (DAPI – blue, dMAb – green). (c) Serum concentration of RSV-F dMAb and Palivizumab after delivery of 200 µg RSV-F dMAb plasmid or 15 mg/kg (clinical dose) Palivizumab measured by ELISA (± SEM, n = 6) dotted line indicates the level of protection (20 µg/ml) for Palivizumab.

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Active immunoprophylaxis with a synthetic DNA-encoded monoclonal anti-respiratory syncytial virus scFv-Fc fusion protein confers protection against infection and durable activity

    doi: 10.1080/21645515.2020.1748979

    Figure Lengend Snippet: Schematic of RSV-F dMAb in-vivo delivery platform : (a) In vivo EP facilitates the delivery of the dMAb to the muscle tissue, myocytes express (b) and secret the encoded mAb (c). mAb is distributed systemically. (b) Example of expression of dMAb in myocytes 3 days after delivery of 50 µg dMAb or pVax (empty expression vector control) to mouse TA (DAPI – blue, dMAb – green). (c) Serum concentration of RSV-F dMAb and Palivizumab after delivery of 200 µg RSV-F dMAb plasmid or 15 mg/kg (clinical dose) Palivizumab measured by ELISA (± SEM, n = 6) dotted line indicates the level of protection (20 µg/ml) for Palivizumab.

    Article Snippet: Antigen binding ELISAAssay plates were coated with 1 µg/well of human RSV F glycoprotein (Sino Biological, Wayne, PA) diluted in 1x DPBS (ThermoFisher) overnight at 4°C.

    Techniques: In Vivo, Expressing, Plasmid Preparation, Concentration Assay, Enzyme-linked Immunosorbent Assay

    Design and in-vitro testing of scFv-Fc RSV-F dMAb construct : (a) Illustration of human IgG1 and scFv-Fc formats. The complete hinge and Fc portions of the molecule are retained in the scFv-Fc format. Yellow line connecting VH and VL of the scFv-Fc molecule represents the (G4 S) 3 linker (b) Plasmid map of RSV-F dMAb (bGH PolyA – bovine growth hormone polyadenylation signal, KanR – kanamycin resistance gene, pUC ori – pUC origin of replication, hCMV promotor – human cytomegalovirus promotor) (c) In vitro expression of RSV-F dMAb. Immuno-fluorescence staining was performed on HEK293 T cells 3 days after in vitro transfection (DAPI – blue, RSV-F dMAb – red) (d) Western-blot analysis of cell-culture supernatant of RSV-F dMAb in vitro transfected HEK293 T cells harvested on day 3 post-transfection (lane 1: IgG-RSV dMAb, lane 2: scFv-Fc RSV-F dMAb).

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Active immunoprophylaxis with a synthetic DNA-encoded monoclonal anti-respiratory syncytial virus scFv-Fc fusion protein confers protection against infection and durable activity

    doi: 10.1080/21645515.2020.1748979

    Figure Lengend Snippet: Design and in-vitro testing of scFv-Fc RSV-F dMAb construct : (a) Illustration of human IgG1 and scFv-Fc formats. The complete hinge and Fc portions of the molecule are retained in the scFv-Fc format. Yellow line connecting VH and VL of the scFv-Fc molecule represents the (G4 S) 3 linker (b) Plasmid map of RSV-F dMAb (bGH PolyA – bovine growth hormone polyadenylation signal, KanR – kanamycin resistance gene, pUC ori – pUC origin of replication, hCMV promotor – human cytomegalovirus promotor) (c) In vitro expression of RSV-F dMAb. Immuno-fluorescence staining was performed on HEK293 T cells 3 days after in vitro transfection (DAPI – blue, RSV-F dMAb – red) (d) Western-blot analysis of cell-culture supernatant of RSV-F dMAb in vitro transfected HEK293 T cells harvested on day 3 post-transfection (lane 1: IgG-RSV dMAb, lane 2: scFv-Fc RSV-F dMAb).

    Article Snippet: Antigen binding ELISAAssay plates were coated with 1 µg/well of human RSV F glycoprotein (Sino Biological, Wayne, PA) diluted in 1x DPBS (ThermoFisher) overnight at 4°C.

    Techniques: In Vitro, Construct, Plasmid Preparation, Expressing, Fluorescence, Staining, Transfection, Western Blot, Cell Culture

    Characterization of RSV-F dMAb in cotton rats: (a) Local expression of dMAb demonstrated by Immuno-fluorescence staining of RSV-F dMAb in cotton rat TA muscle tissue 7 days after delivery of 400 µg RSV-F dMAb pDNA to the tissue site (DAPI – blue; RSV-F dMAb – green; sectioned perpendicular to myocytes) (b) Levels of RSV-F dMAb (ng/ml) was measured for 39 days after IM administration of 800 µg dMAb-plasmid (±SEM, n = 5). (c) Virus neutralization function of in-vivo expressed RSV-F dMAb. Serum samples were harvested and tested 7 days after delivery of 2.4 mg scFv-Fc dMAb-pDNA: neutralizing titer (±SEM, dotted line indicates LOD at serum-dilution of 1/20). (d) Concentration of RSV-F dMAb in BAL samples from treated cotton rats.

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Active immunoprophylaxis with a synthetic DNA-encoded monoclonal anti-respiratory syncytial virus scFv-Fc fusion protein confers protection against infection and durable activity

    doi: 10.1080/21645515.2020.1748979

    Figure Lengend Snippet: Characterization of RSV-F dMAb in cotton rats: (a) Local expression of dMAb demonstrated by Immuno-fluorescence staining of RSV-F dMAb in cotton rat TA muscle tissue 7 days after delivery of 400 µg RSV-F dMAb pDNA to the tissue site (DAPI – blue; RSV-F dMAb – green; sectioned perpendicular to myocytes) (b) Levels of RSV-F dMAb (ng/ml) was measured for 39 days after IM administration of 800 µg dMAb-plasmid (±SEM, n = 5). (c) Virus neutralization function of in-vivo expressed RSV-F dMAb. Serum samples were harvested and tested 7 days after delivery of 2.4 mg scFv-Fc dMAb-pDNA: neutralizing titer (±SEM, dotted line indicates LOD at serum-dilution of 1/20). (d) Concentration of RSV-F dMAb in BAL samples from treated cotton rats.

    Article Snippet: Antigen binding ELISAAssay plates were coated with 1 µg/well of human RSV F glycoprotein (Sino Biological, Wayne, PA) diluted in 1x DPBS (ThermoFisher) overnight at 4°C.

    Techniques: Expressing, Fluorescence, Staining, Plasmid Preparation, Neutralization, In Vivo, Concentration Assay

    In-vivo expression of RSV-F dMAb in BALB/c mice : 200 µg RSV-F dMAb plasmid was administered IM in BALB/c mice (a-d). Serum samples were taken from treated mice before treatment (day 0) and 7 days after treatment to measure (a) Serum expression of RSV-F dMAb. scFv-Fc construct (± SEM, n = 5–8) (b) RSV-F antigen binding of dMAb. RSV-F binding signal of serum samples from treated (squares) and naïve (triangles) mice (± SEM, n = 4) (c) In vitro neutralization of RSV-A virus. Neutralization titer of serum samples from treated (squares) or naïve (triangles) mice (log2 serum dilution of 60% reduction of plaque-formation; ±SEM, n = 3–11, dotted line indicates LOD at serum-dilution of 1/20) (d) Concentration of scFv-Fc RSV-F dMAb in BAL samples 7 days after RSV-dMAb pDNA delivery in treated (squares) or naïve (triangles) mice (mol scFv-Fc dMAb per gram total protein in lavage sample, n(naïve) = 2, n(RSV-F dMAb) = 3).

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Active immunoprophylaxis with a synthetic DNA-encoded monoclonal anti-respiratory syncytial virus scFv-Fc fusion protein confers protection against infection and durable activity

    doi: 10.1080/21645515.2020.1748979

    Figure Lengend Snippet: In-vivo expression of RSV-F dMAb in BALB/c mice : 200 µg RSV-F dMAb plasmid was administered IM in BALB/c mice (a-d). Serum samples were taken from treated mice before treatment (day 0) and 7 days after treatment to measure (a) Serum expression of RSV-F dMAb. scFv-Fc construct (± SEM, n = 5–8) (b) RSV-F antigen binding of dMAb. RSV-F binding signal of serum samples from treated (squares) and naïve (triangles) mice (± SEM, n = 4) (c) In vitro neutralization of RSV-A virus. Neutralization titer of serum samples from treated (squares) or naïve (triangles) mice (log2 serum dilution of 60% reduction of plaque-formation; ±SEM, n = 3–11, dotted line indicates LOD at serum-dilution of 1/20) (d) Concentration of scFv-Fc RSV-F dMAb in BAL samples 7 days after RSV-dMAb pDNA delivery in treated (squares) or naïve (triangles) mice (mol scFv-Fc dMAb per gram total protein in lavage sample, n(naïve) = 2, n(RSV-F dMAb) = 3).

    Article Snippet: Antigen binding ELISAAssay plates were coated with 1 µg/well of human RSV F glycoprotein (Sino Biological, Wayne, PA) diluted in 1x DPBS (ThermoFisher) overnight at 4°C.

    Techniques: In Vivo, Expressing, Mouse Assay, Plasmid Preparation, Construct, Binding Assay, In Vitro, Neutralization, Concentration Assay

    RSV-F dMAb confers protection against LRD after RSV/A challenge of cotton rats: (a) Schematic of cotton rat challenge study: Animals were treated with 2.4 mg RSV-dMAb 7 days before challenge or with an IM injection of 15 mg/kg Palivizumab 1 day before challenge (b) Viral load of cotton rat lung tissue (pfu/g) harvested 5 days after intra-nasal live virus challenge with RSV/A/long (± SEM n = 4–5) (i). RSV Nonstructural protein-1 (NS-1) mRNA levels (log2 and normalized to beta-Actin) of cotton rat lung tissue 5 days after intra-nasal live virus challenge with RSV/A/long (± SEM, n = 4–5, Mann–Whitney non-parametric t-test: p (untreated vs RSV-dMAb) = 0.0159; p (untreated vs Palivizumab) = 0.0079) (ii). (c) Serum levels of Palivizumab and scFv-Fc RSV-F dMAb in cotton rats at the day of challenge (day 0) and 5 days after challenge (± SEM, n = 4–5).

    Journal: Human Vaccines & Immunotherapeutics

    Article Title: Active immunoprophylaxis with a synthetic DNA-encoded monoclonal anti-respiratory syncytial virus scFv-Fc fusion protein confers protection against infection and durable activity

    doi: 10.1080/21645515.2020.1748979

    Figure Lengend Snippet: RSV-F dMAb confers protection against LRD after RSV/A challenge of cotton rats: (a) Schematic of cotton rat challenge study: Animals were treated with 2.4 mg RSV-dMAb 7 days before challenge or with an IM injection of 15 mg/kg Palivizumab 1 day before challenge (b) Viral load of cotton rat lung tissue (pfu/g) harvested 5 days after intra-nasal live virus challenge with RSV/A/long (± SEM n = 4–5) (i). RSV Nonstructural protein-1 (NS-1) mRNA levels (log2 and normalized to beta-Actin) of cotton rat lung tissue 5 days after intra-nasal live virus challenge with RSV/A/long (± SEM, n = 4–5, Mann–Whitney non-parametric t-test: p (untreated vs RSV-dMAb) = 0.0159; p (untreated vs Palivizumab) = 0.0079) (ii). (c) Serum levels of Palivizumab and scFv-Fc RSV-F dMAb in cotton rats at the day of challenge (day 0) and 5 days after challenge (± SEM, n = 4–5).

    Article Snippet: Antigen binding ELISAAssay plates were coated with 1 µg/well of human RSV F glycoprotein (Sino Biological, Wayne, PA) diluted in 1x DPBS (ThermoFisher) overnight at 4°C.

    Techniques: Injection, MANN-WHITNEY

    Intranasal vaccination of mice with NE-RSV results in RSV-specific antibody responses. Mice were immunized with NE-RSV containing 10 5 virus particles at Day 0 and Day 28. In (A), the levels of RSV specific antibodies (IgG+IgM) in serum were determined at weeks 2, 4, 6, 8, and 10 via ELISA using purified RSV protein. In (B), serum samples were serially diluted to obtain endpoints titers. In (C), RSV-specific IgA, IgG2a, IgG1, and IgE were assessed at week 8 via isotype-specific ELISA. In (D), IgG2a and IgG1 antibodies specific for RSV F and G were assessed at week 8 via isotype-specific ELISA using purified RSV F and G glycoproteins. In (E), RSV-specific IgA responses in bronchoalveolar lavage samples (BAL) were assessed using an isotype-specific ELISA. Each time point represents the mean of a minimum of 5 samples +/− SEM, and the experiment was repeated with similar results. * = P

    Journal: PLoS ONE

    Article Title: A Novel Inactivated Intranasal Respiratory Syncytial Virus Vaccine Promotes Viral Clearance without Th2 Associated Vaccine-Enhanced Disease

    doi: 10.1371/journal.pone.0021823

    Figure Lengend Snippet: Intranasal vaccination of mice with NE-RSV results in RSV-specific antibody responses. Mice were immunized with NE-RSV containing 10 5 virus particles at Day 0 and Day 28. In (A), the levels of RSV specific antibodies (IgG+IgM) in serum were determined at weeks 2, 4, 6, 8, and 10 via ELISA using purified RSV protein. In (B), serum samples were serially diluted to obtain endpoints titers. In (C), RSV-specific IgA, IgG2a, IgG1, and IgE were assessed at week 8 via isotype-specific ELISA. In (D), IgG2a and IgG1 antibodies specific for RSV F and G were assessed at week 8 via isotype-specific ELISA using purified RSV F and G glycoproteins. In (E), RSV-specific IgA responses in bronchoalveolar lavage samples (BAL) were assessed using an isotype-specific ELISA. Each time point represents the mean of a minimum of 5 samples +/− SEM, and the experiment was repeated with similar results. * = P

    Article Snippet: RSV F and G specific ELISAs were performed similarly, except that plates were coated with purified recombinant RSV F or RSV G glycoproteins (1.3 ug/ml) expressed in Baculovirus (Sino Biological).

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay, Purification

    Evaluation of serum IgG to recognize antigenic sites I, II, and IV in F protein. Sera were subjected to competitive ELISA by using monoclonal antibody specific to antigenic sites I, II, or IV. The amounts of serum IgG specific for each antigenic epitopes are represented by IC50 resulting in 50% reduction of biding RSV F with competitive monoclonal antibody. The limitation of the IC50 detection was defined as 5. The concentration of serum IgG specific to antigenic site II was calculated based on the result of palivizumab on competitive ELISA. The mean ± 95% CI was estimated by the linear model, and were shown in the graphs. IC50 of antigenic site I-specific IgG in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 26; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 32; adults, n = 23. IC50 of antigenic site I-specific IgG in convalescent phase: 0–3 mo, n = 24; 4–6 mo, n = 27; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 32; adults, n = 23. GMRs C/A of antigenic site I-specific IgG: 0–3 mo, n = 21; 4–6 mo, n = 25; 7–12 mo, n = 26; 13–18 mo, n = 34; 19–36 mo, n = 31; adults, n = 23. IC50 of antigenic site II-specific IgG in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 28; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 33; adults, n = 23. IC50 of antigenic site II-specific IgG in convalescent phase: 0–3 mo, n = 27; 4–6 mo, n = 29; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 33; adults, n = 23. GMRs C/A of antigenic site II-specific IgG: 0–3 mo, n = 24; 4–6 mo, n = 27; 7–12 mo, n = 26; 13–18 mo, n = 34; 19–36 mo, n = 32; adults, n = 23. IC50 of antigenic site IV-specific IgG in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 28; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 31; adults, n = 23. IC50 of antigenic site IV-specific IgG in convalescent phase: 0–3 mo, n = 27; 4–6 mo, n = 29; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 33; adults, n = 23. GMRs C/A of antigenic site IV-specific IgG: 0–3 mo, n = 24; 4–6 mo, n = 27; 7–12 mo, n = 26; 13–18 mo, n = 34; 19–36 mo, n = 31; adults, n = 23. * p

    Journal: EBioMedicine

    Article Title: Age-Specific Profiles of Antibody Responses against Respiratory Syncytial Virus Infection

    doi: 10.1016/j.ebiom.2017.01.014

    Figure Lengend Snippet: Evaluation of serum IgG to recognize antigenic sites I, II, and IV in F protein. Sera were subjected to competitive ELISA by using monoclonal antibody specific to antigenic sites I, II, or IV. The amounts of serum IgG specific for each antigenic epitopes are represented by IC50 resulting in 50% reduction of biding RSV F with competitive monoclonal antibody. The limitation of the IC50 detection was defined as 5. The concentration of serum IgG specific to antigenic site II was calculated based on the result of palivizumab on competitive ELISA. The mean ± 95% CI was estimated by the linear model, and were shown in the graphs. IC50 of antigenic site I-specific IgG in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 26; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 32; adults, n = 23. IC50 of antigenic site I-specific IgG in convalescent phase: 0–3 mo, n = 24; 4–6 mo, n = 27; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 32; adults, n = 23. GMRs C/A of antigenic site I-specific IgG: 0–3 mo, n = 21; 4–6 mo, n = 25; 7–12 mo, n = 26; 13–18 mo, n = 34; 19–36 mo, n = 31; adults, n = 23. IC50 of antigenic site II-specific IgG in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 28; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 33; adults, n = 23. IC50 of antigenic site II-specific IgG in convalescent phase: 0–3 mo, n = 27; 4–6 mo, n = 29; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 33; adults, n = 23. GMRs C/A of antigenic site II-specific IgG: 0–3 mo, n = 24; 4–6 mo, n = 27; 7–12 mo, n = 26; 13–18 mo, n = 34; 19–36 mo, n = 32; adults, n = 23. IC50 of antigenic site IV-specific IgG in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 28; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 31; adults, n = 23. IC50 of antigenic site IV-specific IgG in convalescent phase: 0–3 mo, n = 27; 4–6 mo, n = 29; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 33; adults, n = 23. GMRs C/A of antigenic site IV-specific IgG: 0–3 mo, n = 24; 4–6 mo, n = 27; 7–12 mo, n = 26; 13–18 mo, n = 34; 19–36 mo, n = 31; adults, n = 23. * p

    Article Snippet: 2.4 Enzyme-linked ImmunoSorbent Assay (ELISA) of RSV F and G Protein The 96-well plates were coated with 0.5 μg/mL of F or G protein (SinoBiological Inc.) and serially diluted human IgG protein (Nordic Immunology, Human Standard Serum NOR-01) at 4 °C for 12 h to 18 h. After washing with PBS (-) containing 0.05% Tween 20 (PBST), the plates were treated with the blocking solution (1% BSA in PBST).

    Techniques: Competitive ELISA, Concentration Assay

    Serum antibody response to RSV F protein. The sera were subjected to ELISA coating RSV F protein. Series of detection antibody labeled with HRP were used to acquire the profile of IgG subclasses specific to RSV F protein. A left panel represents concentration of total IgG, IgG1, IgG2 and IgG3 for RSV F in acute phase or convalescent phase, and a right panel does the GMRs C/A of F IgGtotal, IgG1, IgG2, and IgG3. The mean ± 95% CI was estimated by the linear model, and were shown in the graphs. Concentration of antibody in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 28; 7–12 mo, n = 29; 13–18 mo, n = 34; 19–36 mo, n = 35; adults, n = 23. Concentration of antibody in convalescent phase: 0–3 mo, n = 28; 4–6 mo, n = 30; 7–12 mo, n = 33; 13–18 mo, n = 35; 19–36 mo, n = 34; adults, n = 23. GMRs C/A: 0–3 mo, n = 23; 4–6 mo, n = 28; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 34; adults, n = 23. * p

    Journal: EBioMedicine

    Article Title: Age-Specific Profiles of Antibody Responses against Respiratory Syncytial Virus Infection

    doi: 10.1016/j.ebiom.2017.01.014

    Figure Lengend Snippet: Serum antibody response to RSV F protein. The sera were subjected to ELISA coating RSV F protein. Series of detection antibody labeled with HRP were used to acquire the profile of IgG subclasses specific to RSV F protein. A left panel represents concentration of total IgG, IgG1, IgG2 and IgG3 for RSV F in acute phase or convalescent phase, and a right panel does the GMRs C/A of F IgGtotal, IgG1, IgG2, and IgG3. The mean ± 95% CI was estimated by the linear model, and were shown in the graphs. Concentration of antibody in acute phase: 0–3 mo, n = 29; 4–6 mo, n = 28; 7–12 mo, n = 29; 13–18 mo, n = 34; 19–36 mo, n = 35; adults, n = 23. Concentration of antibody in convalescent phase: 0–3 mo, n = 28; 4–6 mo, n = 30; 7–12 mo, n = 33; 13–18 mo, n = 35; 19–36 mo, n = 34; adults, n = 23. GMRs C/A: 0–3 mo, n = 23; 4–6 mo, n = 28; 7–12 mo, n = 28; 13–18 mo, n = 34; 19–36 mo, n = 34; adults, n = 23. * p

    Article Snippet: 2.4 Enzyme-linked ImmunoSorbent Assay (ELISA) of RSV F and G Protein The 96-well plates were coated with 0.5 μg/mL of F or G protein (SinoBiological Inc.) and serially diluted human IgG protein (Nordic Immunology, Human Standard Serum NOR-01) at 4 °C for 12 h to 18 h. After washing with PBS (-) containing 0.05% Tween 20 (PBST), the plates were treated with the blocking solution (1% BSA in PBST).

    Techniques: Enzyme-linked Immunosorbent Assay, Labeling, Concentration Assay

    Antibody affinity to RSV F protein. Sera were subjected to a surface plasmon resonance (SPR) assay. Apparent dissociation rate constants ( k d ) were calculated and shown in different age groups. The mean ± 95% CI was estimated by linear model, and were shown in the graphs. k d in acute phase: 0–3 mo, n = 30; 4–6 mo, n = 29; 7–12 mo, n = 30; 13–18 mo, n = 30; 19–36 mo, n = 32; adults, n = 22. k d in convalescent phase: 0–3 mo, n = 29; 4–6 mo, n = 30; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 33; adults, n = 23. GMRs C/A of F IgG k d : 0–3 mo, n = 25; 4–6 mo, n = 29; 7–12 mo, n = 28; 13–18 mo, n = 30; 19–36 mo, n = 31; adults, n = 22. * p

    Journal: EBioMedicine

    Article Title: Age-Specific Profiles of Antibody Responses against Respiratory Syncytial Virus Infection

    doi: 10.1016/j.ebiom.2017.01.014

    Figure Lengend Snippet: Antibody affinity to RSV F protein. Sera were subjected to a surface plasmon resonance (SPR) assay. Apparent dissociation rate constants ( k d ) were calculated and shown in different age groups. The mean ± 95% CI was estimated by linear model, and were shown in the graphs. k d in acute phase: 0–3 mo, n = 30; 4–6 mo, n = 29; 7–12 mo, n = 30; 13–18 mo, n = 30; 19–36 mo, n = 32; adults, n = 22. k d in convalescent phase: 0–3 mo, n = 29; 4–6 mo, n = 30; 7–12 mo, n = 32; 13–18 mo, n = 35; 19–36 mo, n = 33; adults, n = 23. GMRs C/A of F IgG k d : 0–3 mo, n = 25; 4–6 mo, n = 29; 7–12 mo, n = 28; 13–18 mo, n = 30; 19–36 mo, n = 31; adults, n = 22. * p

    Article Snippet: 2.4 Enzyme-linked ImmunoSorbent Assay (ELISA) of RSV F and G Protein The 96-well plates were coated with 0.5 μg/mL of F or G protein (SinoBiological Inc.) and serially diluted human IgG protein (Nordic Immunology, Human Standard Serum NOR-01) at 4 °C for 12 h to 18 h. After washing with PBS (-) containing 0.05% Tween 20 (PBST), the plates were treated with the blocking solution (1% BSA in PBST).

    Techniques: SPR Assay