anti human tlr4 clone hta125 antibody  (Hycult Biotech)

Journal:

Article Title: Escherichia coli Strain Nissle 1917 Ameliorates Experimental Colitis via Toll-Like Receptor 2- and Toll-Like Receptor 4-Dependent Pathways

doi: 10.1128/IAI.01449-05

Figure Lengend Snippet: E. coli Nissle 1917-conditioned medium increases NF-κB activity via TLR-2- and TLR-4-dependent pathways. After TLR-2 and TLR-4 transfection, E. coli Nissle 1917-conditioned medium increases NF-κB activity. HEK 293 cells were transiently transfected as described in Materials and Methods. At 24 h after transfection cells were stimulated with LP3 or LPS (concentration in ng/ml) or E. coli Nissle 1917 supernatants (EcN) (concentration in μl/ml) or controls in Dulbecco modified Eagle medium. At 20 h after stimulation, cell extracts were prepared for determination of luciferase activity by using a Luciferase Reporter Gene Assay. LP3 was used at 20 ng/ml only. Each bar represents mean ± the SEM value of three separate experiments.

Article Snippet: For flow cytometry, fluorescein isothiocyanate (FITC)-labeled anti-human TLR-2 (clone 6C2) and anti-human TLR-4 (clone HTA125) MAb were purchased from eBioscience.

Techniques: Activity Assay, Transfection, Concentration Assay, Modification, Luciferase, Reporter Gene Assay

Journal:

Article Title: Escherichia coli Strain Nissle 1917 Ameliorates Experimental Colitis via Toll-Like Receptor 2- and Toll-Like Receptor 4-Dependent Pathways

doi: 10.1128/IAI.01449-05

Figure Lengend Snippet: Distinct effect of DSS application and E. coli Nissle 1917 treatment in wt mice and in TLR-2 and TLR-4 knockout mice. In TLR-4 knockout mice, DSS application induced significantly fewer symptoms of colitis compared to wt or TLR-2 knockout mice. wt mice, but not TLR-2 knockout or TLR-4 knockout mice, treated with E. coli Nissle 1917 showed a significant clinical improvement in disease activity, as evaluated with the DAI, compared to mice treated with 0.9% NaCl. Colitis was induced by DSS application in wt mice, TLR-2 knockout mice, and TLR-4 knockout mice. Mice were subsequently treated with an oral or rectal application of 107 E. coli Nissle 1917 (100 μl) or 0.9% NaCl (100 μl) twice daily for 8 days. After sacrifice, the DAI was determined as described in Materials and Methods. Each bar represents mean ± the SEM value of 10 animals. *, P < 0.05 for differences in E. coli Nissle 1917-treated groups versus saline-treated animals.

Article Snippet: For flow cytometry, fluorescein isothiocyanate (FITC)-labeled anti-human TLR-2 (clone 6C2) and anti-human TLR-4 (clone HTA125) MAb were purchased from eBioscience.

Techniques: Knock-Out, Activity Assay

Journal:

Article Title: Escherichia coli Strain Nissle 1917 Ameliorates Experimental Colitis via Toll-Like Receptor 2- and Toll-Like Receptor 4-Dependent Pathways

doi: 10.1128/IAI.01449-05

Figure Lengend Snippet: Differential effect of E. coli Nissle 1917 treatment on inflammation and regeneration in wt mice but not in TLR-2 or TLR-4 knockout mice. In TLR-4 knockout mice DSS application induced significantly less inflammation compared to wt or TLR-2 knockout mice. wt mice, but not TLR-2 knockout or TLR-4 knockout mice, treated with E. coli Nissle 1917 showed significantly less inflammation or crypt damage and more regeneration, as evaluated with the Dieleman index, compared to mice treated with 0.9% NaCl. Colitis was induced by DSS application in wt mice, TLR-2 knockout mice, and TLR-4 knockout mice. Mice were subsequently treated with an oral or rectal application of 107 E. coli Nissle 1917 (100 μl) or 0.9% NaCl (100 μl) twice daily for 8 days. After sacrifice, tissue samples were obtained from the distal 5 cm of the colon and prepared for histological evaluation as described in Materials and Methods. Each bar represents mean ± the SEM value of 10 animals. *, P < 0.05 for differences in E. coli Nissle 1917-treated groups versus saline-treated animals.

Article Snippet: For flow cytometry, fluorescein isothiocyanate (FITC)-labeled anti-human TLR-2 (clone 6C2) and anti-human TLR-4 (clone HTA125) MAb were purchased from eBioscience.

Techniques: Knock-Out

Journal:

Article Title: Escherichia coli Strain Nissle 1917 Ameliorates Experimental Colitis via Toll-Like Receptor 2- and Toll-Like Receptor 4-Dependent Pathways

doi: 10.1128/IAI.01449-05

Figure Lengend Snippet: Differential effect of E. coli Nissle 1917 treatment on mucosal damage in wt mice but not in TLR-2 or TLR-4 knockout mice. wt (A) and TLR-2 knockout (C) mice exposed to 5% DSS developed symptoms of acute colitis to a comparable degree, with a patchy pattern of severe mucosal damage characterized by a loss of crypts, necrosis, and the local influx of inflammatory cells into the lamina propria. (E) In contrast, in TLR-4 knockout mice, DSS application induced significantly fewer symptoms of colitis compared to wt and TLR-2 knockout mice. E. coli Nissle 1917 treatment ameliorated the severity of DSS-induced colitis in wt mice (B) but not in TLR-2 knockout (D) or TLR-4 knockout (F) mice. DSS colitis was induced as described in Materials and Methods, and mice were treated orally with 100 μl of 0.9% NaCl (A, C, and E) or 100 μl of 107 E. coli Nissle 1917 (B, D, and F). Control, non-DSS-treated wt C57BL/6 mice received, by gastric gavage, either 100 μl of isotonic sterile saline (G) or 100 μl of 107 E. coli Nissle 1917 (H). After sacrifice at day 8, tissue samples were obtained from the distal 5 cm of the colon and prepared for histological evaluation as described in Materials and Methods (hematoxylin and eosin stain; original magnifications, ×100).

Article Snippet: For flow cytometry, fluorescein isothiocyanate (FITC)-labeled anti-human TLR-2 (clone 6C2) and anti-human TLR-4 (clone HTA125) MAb were purchased from eBioscience.

Techniques: Knock-Out, H&E Stain

Journal:

Article Title: Escherichia coli Strain Nissle 1917 Ameliorates Experimental Colitis via Toll-Like Receptor 2- and Toll-Like Receptor 4-Dependent Pathways

doi: 10.1128/IAI.01449-05

Figure Lengend Snippet: Distinct effect of DSS application and E. coli Nissle 1917 treatment on MPO activity in wt, TLR-2 knockout, and TLR-4 knockout mice. In TLR-4 knockout mice, DSS application induced significantly less MPO activity in colonic tissue compared to wt or TLR-2 knockout mice. wt mice, but not TLR-2 knockout or TLR-4 knockout mice, treated rectally with 107 E. coli Nissle 1917 showed a significant decrease in MPO activity compared to mice treated with 0.9% NaCl. Colitis was induced by DSS application in wt, TLR-2 knockout, and TLR-4 knockout mice. Mice were subsequently treated with an oral or rectal application of 107 E. coli Nissle 1917 (100 μl) or 0.9% NaCl (100 μl) for 8 days. After sacrifice, the MPO activity was determined as described in Materials and Methods. Each bar represents mean ± the SEM value of 10 animals. *, P < 0.05 for differences in E. coli Nissle 1917-treated groups versus saline-treated animals.

Article Snippet: For flow cytometry, fluorescein isothiocyanate (FITC)-labeled anti-human TLR-2 (clone 6C2) and anti-human TLR-4 (clone HTA125) MAb were purchased from eBioscience.

Techniques: Activity Assay, Knock-Out

Journal:

Article Title: Escherichia coli Strain Nissle 1917 Ameliorates Experimental Colitis via Toll-Like Receptor 2- and Toll-Like Receptor 4-Dependent Pathways

doi: 10.1128/IAI.01449-05

Figure Lengend Snippet: Differential cytokine secretion in wt, TLR-2 knockout, and TLR-4 knockout mice. In wt animals, E. coli Nissle 1917 treatment decreased the secretion of TNF-α, IFN-γ, MCP-1, and IL-10. In TLR-2 knockout mice, only IFN-γ secretion was still high in control animals and reduced by E. coli Nissle 1917, whereas the secretion of TNF-α, MCP-1, and IL-10 was reduced compared to wt controls and not altered by E. coli Nissle 1917 treatment. In TLR-4 knockout mice, TNF-α, IFN-γ, IL-6, and IL-10 secretion was nearly abolished, and MCP-1 secretion was dramatically reduced compared to wt animals. A total of 105 freshly isolated PBT from wt, TLR-2 knockout, and TLR-4 knockout mice, treated orally or rectally with 0.9% NaCl or 107 E. coli Nissle 1917, were cultured for 3 days in the presence of anti-CD3 and CD28 MAbs. Supernatants were then collected, and cytokine secretion was analyzed by using a cytometric bead array assay. Each bar represents the mean ± the SEM value of 10 animals. *, P < 0.05 for differences in E. coli Nissle 1917-treated groups versus saline-treated animals.

Article Snippet: For flow cytometry, fluorescein isothiocyanate (FITC)-labeled anti-human TLR-2 (clone 6C2) and anti-human TLR-4 (clone HTA125) MAb were purchased from eBioscience.

Techniques: Knock-Out, Isolation, Cell Culture