recombinant murine xcl1  (R&D Systems)

Journal: Frontiers in immunology

Article Title: Blockade of XCL1/Lymphotactin Ameliorates Severity of Periprosthetic Osteolysis Triggered by Polyethylene-Particles.

doi: 10.3389/fimmu.2020.01720

Figure Lengend Snippet: FIGURE 1 | Expression of XCL1 and XCR1 in periprosthetic tissues. (A–C) Detection of XCR1 in human tissues around loosening hip-implant by the immunofluorescence test. Deparaffinized sections were stained to observe F4/80, CD68, iNOS (green), XCR1 (red), and cell nuclei (blue). Scale bars are 100 µm. Representative images exhibit the sectioned tissues from three patients. (D) Detection of XCL1 in synovial fluid from same patients by Western blotting analysis. (E) Gene expressions of XCL1 and XCR 1 in calvarial bone tissues in a murine osteolysis calvarial model. Results represent the means of relative expression values ± SEM of three mice. *indicates a significant difference as determined by the Student t-test (p ≤0.05).

Article Snippet: For the XCL1-induced osteolysis model, collagen sponges of the single layer type (PELNAC, Tokyo, Japan) were soaked in 1 or 2 μg of recombinant mouse XCL1 (R&D system) or PBS (n = 6 for each group) and transplanted subcutaneously onto the calvariae for 7 days (13).

Techniques: Expressing, Staining, Western Blot

Journal: Frontiers in immunology

Article Title: Blockade of XCL1/Lymphotactin Ameliorates Severity of Periprosthetic Osteolysis Triggered by Polyethylene-Particles.

doi: 10.3389/fimmu.2020.01720

Figure Lengend Snippet: FIGURE 2 | Administration of XCL1 exaggerates osteolytic lesions in a polyethylene-particles-induced osteolysis model. (A) Representative images for micro-CT of calvariae. The right panel shows quantification of the lytic area on the calvarial bone tissues of mice. Results represent the means ± SEM of seven mice. *represents the significance determined by one-way ANOVA, followed by a Tukey’s multiple-comparison procedure. Arrows indicate osteolytic lesions. (B) Representative images for histological analyses of bone sections stained by H&E and TRAP. Scale bar is 100 µm. Arrows indicate bone lesions. (C) Cell count of inflammatory cells in calvarial bone sections. (D) Quantification of TRAP-stained areas in calvarial bone sections. The results represent the means ± SEM for three mice. *indicates a significant difference, as determined by one-way ANOVA, followed by the Tukey’s multiple-comparison procedure (p ≤0.05).

Article Snippet: For the XCL1-induced osteolysis model, collagen sponges of the single layer type (PELNAC, Tokyo, Japan) were soaked in 1 or 2 μg of recombinant mouse XCL1 (R&D system) or PBS (n = 6 for each group) and transplanted subcutaneously onto the calvariae for 7 days (13).

Techniques: Micro-CT, Comparison, Staining, Cell Counting

Journal: Frontiers in immunology

Article Title: Blockade of XCL1/Lymphotactin Ameliorates Severity of Periprosthetic Osteolysis Triggered by Polyethylene-Particles.

doi: 10.3389/fimmu.2020.01720

Figure Lengend Snippet: FIGURE 3 | Blockade of XCL1/lymphotactin by neutralizing antibody ameliorates severity of osteolysis triggered by polyethylene-particles in murine model. (A) Quantification of the lytic area on the calvarial bone tissues of mice determined by micro-CT. Left panel shows representative images of calvariae. Results represent the means ± SEM of seven mice. *represents the significance determined by one-way ANOVA, followed by a Tukey’s multiple-comparison procedure. Arrows indicate osteolytic lesions. (B) Histological analyses of bone sections stained by H&E and TRAP. Scale bar is 100 µm. (C) Cell count of inflammatory cells in calvarial bone sections. (D) Quantification of TRAP-stained areas in calvarial bone sections. Arrows indicate bone lesions. The results represent the means ± SEM for four mice. *indicates a significant difference, as determined the Student t-test (p ≤0.05).

Article Snippet: For the XCL1-induced osteolysis model, collagen sponges of the single layer type (PELNAC, Tokyo, Japan) were soaked in 1 or 2 μg of recombinant mouse XCL1 (R&D system) or PBS (n = 6 for each group) and transplanted subcutaneously onto the calvariae for 7 days (13).

Techniques: Micro-CT, Comparison, Staining, Cell Counting

Journal: Frontiers in immunology

Article Title: Blockade of XCL1/Lymphotactin Ameliorates Severity of Periprosthetic Osteolysis Triggered by Polyethylene-Particles.

doi: 10.3389/fimmu.2020.01720

Figure Lengend Snippet: FIGURE 4 | Sponge-soaked protein-induced murine model. (A) Representative images for micro-CT of calvariae. Arrows indicate osteolytic lesions. (B) Quantification of the lytic area on the calvarial bone tissues of mice determined by micro-CT. Results represent the means ± SEM of six mice. *represents the significance determined by one-way ANOVA, followed by a Tukey’s multiple-comparison procedure. (C) Representative images for histological analyses of bone sections stained by H&E and TRAP. Scale bar is 100 µm. Arrows indicate bone lesions. (D) Quantification of TRAP-stained areas in calvarial bone sections. Results represent the means ± SEM of three mice. *indicates a significant difference, as determined by one-way ANOVA, followed by the Tukey’s multiple-comparison procedure (p ≤0.05). (E) Heat map for the gene expression of inflammatory and osteoclast marker genes in bone tissues. Calvarial bone tissues were harvested for the analysis of gene expressions after the implantation of XCL1-soaked sponges. Scale bar (Log2) represents the means of relative expression values of each target gene after normalizing with the GAPDH ± SEM of three mice. *indicates a significant difference, as determined by the t-test (p ≤0.05).

Article Snippet: For the XCL1-induced osteolysis model, collagen sponges of the single layer type (PELNAC, Tokyo, Japan) were soaked in 1 or 2 μg of recombinant mouse XCL1 (R&D system) or PBS (n = 6 for each group) and transplanted subcutaneously onto the calvariae for 7 days (13).

Techniques: Micro-CT, Comparison, Staining, Gene Expression, Marker, Expressing

Journal: Frontiers in immunology

Article Title: Blockade of XCL1/Lymphotactin Ameliorates Severity of Periprosthetic Osteolysis Triggered by Polyethylene-Particles.

doi: 10.3389/fimmu.2020.01720

Figure Lengend Snippet: FIGURE 5 | Effect of XCL1 on osteoclast differentiation and bone resorption. (A) Count of TRAP-positive cells in RANKL-stimulated monocytes in the presence or absence of XCL1. Left panel shows representative images for cells stained by TRAP. Results represent the means ± SEM of triplicates. *indicates a significant difference, as determined by the Tukey’s multiple comparisons test (p ≤0.05). (B) Actin ring staining assay for cells in RANKL-stimulated monocytes in the presence or absence of XCL1. (C) Quantification of the bone resorbed areas on dentin slices. Results represent the means ± SEM of values from three dentin slices. Left panel shows representative images for the resorbed areas. Scale bars are 200 µm.

Article Snippet: For the XCL1-induced osteolysis model, collagen sponges of the single layer type (PELNAC, Tokyo, Japan) were soaked in 1 or 2 μg of recombinant mouse XCL1 (R&D system) or PBS (n = 6 for each group) and transplanted subcutaneously onto the calvariae for 7 days (13).

Techniques: Staining

Journal: Frontiers in immunology

Article Title: Blockade of XCL1/Lymphotactin Ameliorates Severity of Periprosthetic Osteolysis Triggered by Polyethylene-Particles.

doi: 10.3389/fimmu.2020.01720

Figure Lengend Snippet: FIGURE 7 | The summary of the current study. XCL1 promotes cells infiltrate, inflammatory response, and osteoclast differentiation leading to aseptic loosening. Scale bars are indicated on histological images. Blocking of the XCL1 might be a potent therapeutic target for this clinical problem.

Article Snippet: For the XCL1-induced osteolysis model, collagen sponges of the single layer type (PELNAC, Tokyo, Japan) were soaked in 1 or 2 μg of recombinant mouse XCL1 (R&D system) or PBS (n = 6 for each group) and transplanted subcutaneously onto the calvariae for 7 days (13).

Techniques: Blocking Assay

Journal: Frontiers in immunology

Article Title: Blockade of XCL1/Lymphotactin Ameliorates Severity of Periprosthetic Osteolysis Triggered by Polyethylene-Particles.

doi: 10.3389/fimmu.2020.01720

Figure Lengend Snippet: FIGURE 6 | Effect of XCL1 on osteoblast activation and function. (A) Effect of XCL1 on the gene expression of inflammatory and osteoclastogenic factors in differentiated osteoblasts. Results represent the means ± SEM of triplicates and *indicates a significant difference, as determined by the Tukey’s multiple comparisons test (p ≤0.05). (B) Effects of stimulation by recombinant proteins on NFKB, SAPK/JNK, P42/p44-MAPK (Erk1/2), and P38- activities. Human osteoblasts were cultured in a differentiation medium supplemented with either XCL1 or TNFα (positive control) and harvested for the gene expression analysis by qRT-PCR or for protein analysis by western blotting.

Article Snippet: For the XCL1-induced osteolysis model, collagen sponges of the single layer type (PELNAC, Tokyo, Japan) were soaked in 1 or 2 μg of recombinant mouse XCL1 (R&D system) or PBS (n = 6 for each group) and transplanted subcutaneously onto the calvariae for 7 days (13).

Techniques: Activation Assay, Gene Expression, Recombinant, Cell Culture, Positive Control, Quantitative RT-PCR, Western Blot