human cx3cl1 chemokine domain blocking antibody  (R&D Systems)

Journal: Scientific Reports

Article Title: Indicators of responsiveness to immune checkpoint inhibitors

doi: 10.1038/s41598-017-01000-2

Figure Lengend Snippet: Responding tumors show increased T-cells and Chemokines prior to treatment. ( a , c ) Representative CD8 + and CD3 + IHC staining of the invasive tumor margin and intratumoral region in pretreatment metastatic melanoma tumors (responding N = 4, non-responding N = 4). Tumor compartments were demarcated by a dermatopathologist. ( b , d ) Average CD8 + and CD3 + cell counts for responding and non-responding tumors’ compartments. T-cell counts were generated by averaging the counts of 10 randomly selected fields at 20x objective for each tumor compartment (10 invasive margin; 10 intratumoral). Individual tumor counts can be found in Extended Data Fig. 1. RIM = Responding invasive margin; NRIM = Non-responding invasive margin; RIT = Responding intratumoral; NRIT = Non-responding intratumoral. ( e ) Reverse western assay with the human chemokine antibody arrays (R&D Systems). Results are ratios of summed intensities of responding and non-responding tumors, ratios >2 were defined as a significant change. Chemokine signaling was higher in responding tumors with 10 of 31 chemokines showing >2 fold change. All error bars denote the s.e.m.

Article Snippet: RIM = Responding invasive margin; NRIM = Non-responding invasive margin; RIT = Responding intratumoral; NRIT = Non-responding intratumoral. ( e ) Reverse western assay with the human chemokine antibody arrays (R&D Systems).

Techniques: Immunohistochemistry, Generated, Western Blot

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Arterial and Venous Endothelia Display Differential Functional Fractalkine (CX 3 CL1) Expression by Angiotensin-II

doi: 10.1161/atvbaha.112.254870

Figure Lengend Snippet: Figure 1. Effect of angiotensin-II (Ang-II) on arteriolar leukocyte adhesion (A), venular leukocyte adhesion (B), and venular leu kocyte emigration (C) on mouse cremasteric microcirculation. CX3CL1 was expressed in the cremasteric arterioles and postcapil lary venules of Ang-II–injected animals (D). Results are mean±SEM of n=4 to 6 animals per group. **P<0.01 relative to vehicle-treated mice; +P<0.05 relative to CX3CR1−/+-treated mice.

Article Snippet: The PE-conjugated mouse monoclonal anti-human CX3CL1 (clone 51637), the CFS-conjugated mouse monoclonal anti-human CX3CR1 (clone 528728) and the mouse monoclonal anti-human CX3CL1 (clone 81506) were purchased from R&D Systems (Abingdon, UK).

Techniques: Injection

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Arterial and Venous Endothelia Display Differential Functional Fractalkine (CX 3 CL1) Expression by Angiotensin-II

doi: 10.1161/atvbaha.112.254870

Figure Lengend Snippet: Figure 2. Angiotensin-II (Ang-II) induces CX3CL1 mRNA (A and B) and protein (flow cytometry; C and D) expression in human umbilical arterial endothelial cells (HUAEC) and human umbilical vein endothelial cells (HUVEC). A neutralizing antibody against CX3CL1 function inhibited the recruitment of mononuclear leukocytes to Ang-II–stimulated HUAEC but not HUVEC (E and F). Results are mean±SEM of n=4 to 6 independent experiments. *P<0.05 or **P<0.01 relative to values in the medium group; +P<0.05 relative to values in the Ang-II group. CX3CL1 expression was visualized in nonpermeabilized HUAEC and HUVEC by immunofluorescence (green). Nuclei were coun terstained with 4′,6-diamidino-2-phenylindole. Results are representative of n=5 independent experiments. In the flow chamber assay, results are mean±SEM of n= 5 to 7 independent experiments. *P<0.05 or **P<0.01 relative to values in the medium group; +P<0.05 rela tive to Ang-II+MOPC21 group. TNFα indicates tumor necrosis factor-α.

Article Snippet: The PE-conjugated mouse monoclonal anti-human CX3CL1 (clone 51637), the CFS-conjugated mouse monoclonal anti-human CX3CR1 (clone 528728) and the mouse monoclonal anti-human CX3CL1 (clone 81506) were purchased from R&D Systems (Abingdon, UK).

Techniques: Flow Cytometry, Expressing, Immunofluorescence, Boyden Chamber Assay

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Arterial and Venous Endothelia Display Differential Functional Fractalkine (CX 3 CL1) Expression by Angiotensin-II

doi: 10.1161/atvbaha.112.254870

Figure Lengend Snippet: Figure 4. Angiotensin-II (Ang-II)–induced CX3CL1 expression (A) and mononuclear cell arrest (B) are inhibited by apocynin but not by allopurinol. Nox 5 but not Nox 2 or Nox 4 small interfering RNA (siRNA) inhibits Ang-II–induced CX3CL1 expression and mononuclear cell arrest in human umbilical arterial endothelial cells (HUAEC; C–H). HUAEC were stimulated with 1 µmol/L Ang-II for 24 hours. Some cells were pretreated with apocynine (30 μmol/L) or allopurinol (100 μmol/L) 1 hour before Ang-II stimulation. Results are the mean±SEM of n=7 independent experiments. **P<0.01 relative to values in the medium group; ++P<0.01 relative to values in the Ang-II–stimulated group. In endothelial cells transfected with Nox 2, Nox 4 or Nox 5 siRNA or control siRNA, results are mean±SEM of n=6 independent experiments. *P<0.05 or **P<0.01 relative to values in the medium group; +P<0.05 or ++P<0.01 relative to values in Ang-II group in control siRNA–transfected cells. XO indicates xantine oxidase.

Article Snippet: The PE-conjugated mouse monoclonal anti-human CX3CL1 (clone 51637), the CFS-conjugated mouse monoclonal anti-human CX3CR1 (clone 528728) and the mouse monoclonal anti-human CX3CL1 (clone 81506) were purchased from R&D Systems (Abingdon, UK).

Techniques: Expressing, Small Interfering RNA, Transfection, Control

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Arterial and Venous Endothelia Display Differential Functional Fractalkine (CX 3 CL1) Expression by Angiotensin-II

doi: 10.1161/atvbaha.112.254870

Figure Lengend Snippet: Figure 3. Tumor necrosis factor-α (TNFα) mRNA (A) and pro tein (B) expression in human umbilical arterial endothelial cells (HUAEC) is abolished in HUAEC transfected with small interfer ing RNA (siRNA) targeting TNFα. TNFα siRNA inhibits Ang-II– induced CX3CL1 expression (C) and mononuclear–endothelial cell interactions (D) in HUAEC. Results are mean±SEM of n= 6 independent experiments. *P<0.05 or **P<0.01 relative to values in the medium group; +P<0.05 or ++P<0.01 relative to their respective group in control siRNA–transfected cells. MFI indicates mean fluorescence intensity.

Article Snippet: The PE-conjugated mouse monoclonal anti-human CX3CL1 (clone 51637), the CFS-conjugated mouse monoclonal anti-human CX3CR1 (clone 528728) and the mouse monoclonal anti-human CX3CL1 (clone 81506) were purchased from R&D Systems (Abingdon, UK).

Techniques: Expressing, Transfection, Control, Fluorescence

Journal: Arteriosclerosis, Thrombosis, and Vascular Biology

Article Title: Arterial and Venous Endothelia Display Differential Functional Fractalkine (CX 3 CL1) Expression by Angiotensin-II

doi: 10.1161/atvbaha.112.254870

Figure Lengend Snippet: Figure 6. Combined stimulation with tumor necrosis factor-α (TNFα), interferon γ (IFNγ), and angiotensin-II (Ang-II) promotes increased CX3CL1 expression (A) and mononuclear–endothelial cell interactions (B). Cells were treated with Ang-II (1 µmol/L), TNFα (20 ng/mL), IFNγ (20 ng/mL), Ang-II+TNFα, Ang-II+IFNγ, TNFα+IFNγ, or Ang-II+TNFα+IFNγ for 24 hours. Results are the mean±SEM of n=6 to 9 independent experiments (A) and n=6 to 7 independent experiments (B). *P<0.05 or **P<0.01 relative to values in the medium group; +P<0.05 or ++P<0.01 relative to val ues in the Ang-II group; &P<0.05 or &&P<0.01 relative to values in the TNFα group; ##P<0.01 relative to values in the IFNγ group.

Article Snippet: The PE-conjugated mouse monoclonal anti-human CX3CL1 (clone 51637), the CFS-conjugated mouse monoclonal anti-human CX3CR1 (clone 528728) and the mouse monoclonal anti-human CX3CL1 (clone 81506) were purchased from R&D Systems (Abingdon, UK).

Techniques: Expressing

Journal: Arthritis Research & Therapy

Article Title: Serum level of soluble CX3CL1/fractalkine is elevated in patients with polymyositis and dermatomyositis, which is correlated with disease activity

doi: 10.1186/ar3761

Figure Lengend Snippet: CX3CL1 expressions in affected muscle of patients with polymyositis and patients with dermatomyositis . Muscle specimens from (A, B) controls, (C, D, G, H) polymyositis (PM) patients, and (E, F) dermatomyositis (DM) patients were stained with anti-CX3CL1 mAb (A, C, E, G) or an isotype-matched control mAb (B, D, F, H). Original magnification, ×200.

Article Snippet: The sections were incubated with 2 μg/ml mouse anti-CX3CL1 mAb (81513; R&D Systems) or 2 μg/ml rabbit anti-CX3CR1 antibody (Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Staining, Control

Journal: Arthritis Research & Therapy

Article Title: Serum level of soluble CX3CL1/fractalkine is elevated in patients with polymyositis and dermatomyositis, which is correlated with disease activity

doi: 10.1186/ar3761

Figure Lengend Snippet: Expression of CX3CL1 and CX3CR1 in the muscle tissue from each of the patients

Article Snippet: The sections were incubated with 2 μg/ml mouse anti-CX3CL1 mAb (81513; R&D Systems) or 2 μg/ml rabbit anti-CX3CR1 antibody (Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Expressing

Journal: Arthritis Research & Therapy

Article Title: Serum level of soluble CX3CL1/fractalkine is elevated in patients with polymyositis and dermatomyositis, which is correlated with disease activity

doi: 10.1186/ar3761

Figure Lengend Snippet: CX3CL1 and CX3CR1 expressions in lung with ILD in polymyositis patients . Lung specimens from (A, B, G, H) controls and (C to F and I to L) polymyositis (PM) patients with ILD were stained with anti-CX3CL1 mAb (A, C, E), anti-CX3CR1 antibody (G, I, K), or isotype-matched control antibodies (B, D, F, H, J, L). Arrows indicate the positive infiltrated mononuclear cells (I), alveolar macrophages (M), vascular endothelial cells (V), and epithelial cells (E). Original magnification, ×400.

Article Snippet: The sections were incubated with 2 μg/ml mouse anti-CX3CL1 mAb (81513; R&D Systems) or 2 μg/ml rabbit anti-CX3CR1 antibody (Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Staining, Control

Journal: Arthritis Research & Therapy

Article Title: Serum level of soluble CX3CL1/fractalkine is elevated in patients with polymyositis and dermatomyositis, which is correlated with disease activity

doi: 10.1186/ar3761

Figure Lengend Snippet: Expression of CX3CL1 and CX3CR1 in the lung tissue from each of the patients

Article Snippet: The sections were incubated with 2 μg/ml mouse anti-CX3CL1 mAb (81513; R&D Systems) or 2 μg/ml rabbit anti-CX3CR1 antibody (Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Expressing

Journal: Arthritis Research & Therapy

Article Title: Serum level of soluble CX3CL1/fractalkine is elevated in patients with polymyositis and dermatomyositis, which is correlated with disease activity

doi: 10.1186/ar3761

Figure Lengend Snippet: Serum level of CX3CL1 in patients with polymyositis, patients with dermatomyositis and healthy controls . Serum samples were collected from polymyositis (PM) patients ( n = 7), dermatomyositis (DM) patients ( n = 22) and healthy controls ( n = 20). (A) Concentration of serum soluble CX3CL1 measured by ELISA. Values are mean ± standard error of the mean. (B) CX3CL1 levels before and after treatment in PM and DM patients ( n = 14; three PM patients and 11 DM patients). Symbols joined by a solid line represent data from an individual subject.

Article Snippet: The sections were incubated with 2 μg/ml mouse anti-CX3CL1 mAb (81513; R&D Systems) or 2 μg/ml rabbit anti-CX3CR1 antibody (Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay

Journal: Arthritis Research & Therapy

Article Title: Serum level of soluble CX3CL1/fractalkine is elevated in patients with polymyositis and dermatomyositis, which is correlated with disease activity

doi: 10.1186/ar3761

Figure Lengend Snippet: Correlation of serum CX3CL1 level and the disease activity of polymyositis and dermatomyositis . Correlations of the level of CX3CL1 with (A) serum creatinine kinase (CK) level ( n = 29), (B) manual muscle testing (MMT) score ( n = 18) in patients with polymyositis (PM) and patients with dermatomyositis (DM), and (C) the alveolar-arterial oxygen pressure difference (AaDO 2 ) ( n = 19) in patients with PM and with DM complicated by ILD. Each point represents an individual sample.

Article Snippet: The sections were incubated with 2 μg/ml mouse anti-CX3CL1 mAb (81513; R&D Systems) or 2 μg/ml rabbit anti-CX3CR1 antibody (Abcam, Cambridge, MA, USA) at 4°C overnight.

Techniques: Activity Assay