Journal: Cells

Article Title: Human Lung-Resident Macrophages Express and Are Targets of Thymic Stromal Lymphopoietin in the Tumor Microenvironment

doi: 10.3390/cells10082012

Figure Lengend Snippet: Constitutive expression of TSLP system in human lung macrophages (HLMs). The constitutive expression of sfTSLP, lfTSLP, TSLPR and IL-7Rα mRNAs was evaluated by quantitative RT-PCR in highly purified HLMs (4.5 × 10 6 cells/well) ( A ). Data are mean ± SD of 6 independent experiments obtained from different patients. Cytocentrifuge preparations of HLMs were immunohistochemically stained for TSLP ( C ), TSLPR ( D ), and IL-7Rα ( E ) with specific primary antibodies or in absence ( B ) of primary antibody (CTRL) as described in Materials and Methods. Microscope magnification 60×. Results are representative of 6 independent experiments obtained from different patients. * p < 0.01 when compared to lfTSLP, TSLPR, IL-7Rα. px: pixels.

Article Snippet: Paraffin-embedded sections were processed for immunohistochemistry by peroxidase anti-peroxidase method using as primary antibody rabbit polyclonal anti-TSLP antibody (1:100) (PA5-78610), rabbit polyclonal antibody anti-TSLP Receptor (PA5-203789, or rabbit polyclonal antibody anti-IL-7Rα (1:100) (PA5-97870) (Invitrogen, Thermo Fisher Scientific, Monza, Italy).

Techniques: Expressing, Quantitative RT-PCR, Purification, Staining, Microscopy

Journal: Cells

Article Title: Human Lung-Resident Macrophages Express and Are Targets of Thymic Stromal Lymphopoietin in the Tumor Microenvironment

doi: 10.3390/cells10082012

Figure Lengend Snippet: Effects of IL-13 and IL-4, alone or in combination, and of LPS on TSLP system in HLMs. Highly purified HLMs (1.5 × 10 5 cells/well) were incubated (16 h, 37 °C) in the absence (CTRL) or in the presence of IL-13 (10 ng/mL), IL-4 (10 ng/mL) or their combination, or LPS (100 ng/mL ( A , B ). At the end of incubation, TSLP ( A ) and CXCL8 ( B ) proteins in supernatants were evaluated by ELISA. In parallel experiments, HLM (4.5 × 10 6 cells/well) were incubated (6 h, 37 °C) in the absence (CTRL) or in presence of IL-13 (10 ng/mL), IL-4 (10 ng/mL) or their combination, or LPS (100 ng/mL). At the end of incubation, sfTSLP ( C ), lfTSLP ( D ), TSLPR ( E ), and IL-7Rα ( F ) mRNAs were determined by quantitative RT-PCR. Data are mean ± SD of 6 independent experiments obtained from different patients. * p < 0.01, ** p < 0.001 and § p < 0.0001 vs. CTRL.

Article Snippet: Paraffin-embedded sections were processed for immunohistochemistry by peroxidase anti-peroxidase method using as primary antibody rabbit polyclonal anti-TSLP antibody (1:100) (PA5-78610), rabbit polyclonal antibody anti-TSLP Receptor (PA5-203789, or rabbit polyclonal antibody anti-IL-7Rα (1:100) (PA5-97870) (Invitrogen, Thermo Fisher Scientific, Monza, Italy).

Techniques: Purification, Incubation, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR

Journal: Cells

Article Title: Human Lung-Resident Macrophages Express and Are Targets of Thymic Stromal Lymphopoietin in the Tumor Microenvironment

doi: 10.3390/cells10082012

Figure Lengend Snippet: Effects of IL-13 and IL-4, alone or in combination, on TSLP and CXCL8 release from LPS-activated HLMs. Highly purified HLMs (1.5 × 10 5 cells/well) were preincubated (10 min, 37 °C) with IL-13 (10 ng/mL) or IL-4 (10 ng/mL), alone or in combination, before the stimulation with LPS (100 ng/mL). TSLP ( A ) and CXCL8 ( B ) proteins in supernatants were evaluated by ELISA. Data are mean ± SD of 6 independent experiments obtained from different patients. * p < 0.01, ** p < 0.001 and § p < 0.0001 vs. LPS alone.

Article Snippet: Paraffin-embedded sections were processed for immunohistochemistry by peroxidase anti-peroxidase method using as primary antibody rabbit polyclonal anti-TSLP antibody (1:100) (PA5-78610), rabbit polyclonal antibody anti-TSLP Receptor (PA5-203789, or rabbit polyclonal antibody anti-IL-7Rα (1:100) (PA5-97870) (Invitrogen, Thermo Fisher Scientific, Monza, Italy).

Techniques: Purification, Enzyme-linked Immunosorbent Assay

Journal: Cells

Article Title: Human Lung-Resident Macrophages Express and Are Targets of Thymic Stromal Lymphopoietin in the Tumor Microenvironment

doi: 10.3390/cells10082012

Figure Lengend Snippet: Constitutive expression of sfTSLP, lfTSLP, TSLPR, and IL-7Rα mRNAs and intracellular concentration of TSLP protein in MDMs and monocytes. MDMs (4.5 × 10 6 cells/well) ( A ) and freshly purified monocytes (4.5 × 10 6 cells/well) ( B ) were lysed with RNA lysis buffer to evaluate the expression of sfTSLP, lfTSLP, TSLPR and IL-7Rα mRNAs by quantitative RT-PCR ( A , B ). Total TSLP intracellular concentrations in MDMs and monocytes were evaluated by ELISA ( C ). Data are mean ± SD of 6 independent experiments obtained from different healthy donors. * p < 0.05 when compared to lfTSLP, TSLPR, IL-7Rα. ** p < 0.01 when compared to lfTSLP, TSLPR, IL-7Rα.

Article Snippet: Paraffin-embedded sections were processed for immunohistochemistry by peroxidase anti-peroxidase method using as primary antibody rabbit polyclonal anti-TSLP antibody (1:100) (PA5-78610), rabbit polyclonal antibody anti-TSLP Receptor (PA5-203789, or rabbit polyclonal antibody anti-IL-7Rα (1:100) (PA5-97870) (Invitrogen, Thermo Fisher Scientific, Monza, Italy).

Techniques: Expressing, Concentration Assay, Purification, Lysis, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay

Journal: Cells

Article Title: Human Lung-Resident Macrophages Express and Are Targets of Thymic Stromal Lymphopoietin in the Tumor Microenvironment

doi: 10.3390/cells10082012

Figure Lengend Snippet: Effects of IL-13, IL-4, alone or in combination, and of LPS on TSLP production by MDMs and monocytes. MDMs (1.5 × 10 5 cells/well) ( A ) and monocytes (1.5 × 10 5 cells/well) ( B ) were incubated (16 h, 37 °C) in the presence of IL-13 (10 ng/mL), IL-4 (10 ng/mL), alone or in combination, or LPS (100 ng/mL). In parallel experiments, MDMs (1.5 × 10 5 cells/well) ( C ) and monocytes (1.5 × 10 5 cells/well) ( D ) were preincubated (10 min, 37 °C) with IL-13 (10 ng/mL) or IL-4 (10 ng/mL), alone or in combination, before the stimulation with LPS (100 ng/mL). Incubation continued for 16 h at 37 °C. In both groups of experiments, at the end of incubations TSLP concentrations in supernatants were evaluated by ELISA. Data are mean ± SD of 6 independent experiments obtained from different healthy donors. * p < 0.01, ** p < 0.001 and § p < 0.0001 vs. CTRL ( A , B ) and vs. LPS ( C , D ).

Article Snippet: Paraffin-embedded sections were processed for immunohistochemistry by peroxidase anti-peroxidase method using as primary antibody rabbit polyclonal anti-TSLP antibody (1:100) (PA5-78610), rabbit polyclonal antibody anti-TSLP Receptor (PA5-203789, or rabbit polyclonal antibody anti-IL-7Rα (1:100) (PA5-97870) (Invitrogen, Thermo Fisher Scientific, Monza, Italy).

Techniques: Incubation, Enzyme-linked Immunosorbent Assay

Journal: Cells

Article Title: Human Lung-Resident Macrophages Express and Are Targets of Thymic Stromal Lymphopoietin in the Tumor Microenvironment

doi: 10.3390/cells10082012

Figure Lengend Snippet: Effects of TSLP on the release and expression of inflammatory, angiogenic and lymphangiogenic mediators from HLMs. HLMs (1.5 × 10 5 cells/well) were incubated (16 h, 37 °C) in the absence (CTRL) or in the presence of TSLP (5 ng/mL). At the end of the incubation, TNF-α ( A ), VEGF-A ( B ), VEGF-C ( C ) and ANGPT2 ( D ), and concentrations in the supernatants were evaluated by ELISA. In parallel experiments, HLMs (4.5 × 10 5 cells/well) were incubated (6 h, 37 °C) in the absence (CTRL) or in the presence of TSLP (5 ng/mL). TNF-α ( E ), VEGF-A ( F ), VEGF-C ( G ), and ANGPT2 ( H ) mRNAs were determined by quantitative RT-PRC. Data are mean ± SD of 4 independent experiments obtained from different patients. § p < 0.0001 vs. control (CTRL). * p < 0.05 vs. control; ** p < 0.01 vs. control (CTRL).

Article Snippet: Paraffin-embedded sections were processed for immunohistochemistry by peroxidase anti-peroxidase method using as primary antibody rabbit polyclonal anti-TSLP antibody (1:100) (PA5-78610), rabbit polyclonal antibody anti-TSLP Receptor (PA5-203789, or rabbit polyclonal antibody anti-IL-7Rα (1:100) (PA5-97870) (Invitrogen, Thermo Fisher Scientific, Monza, Italy).

Techniques: Expressing, Incubation, Enzyme-linked Immunosorbent Assay, Control

Journal: Cells

Article Title: Human Lung-Resident Macrophages Express and Are Targets of Thymic Stromal Lymphopoietin in the Tumor Microenvironment

doi: 10.3390/cells10082012

Figure Lengend Snippet: Expression of TSLP, TSLPR, and IL-7Rα in peritumoral and intratumoral human lung cancer by immunohistochemistry. Immunohistochemical staining for TSLP, TSLPR, and IL-7Rα in peritumoral and intratumoral human lung cancer. In the control (CTRL), the primary antibody was omitted. Microscope magnification 20×. This experiment is representative of 5 experiments obtained from different patients.

Article Snippet: Paraffin-embedded sections were processed for immunohistochemistry by peroxidase anti-peroxidase method using as primary antibody rabbit polyclonal anti-TSLP antibody (1:100) (PA5-78610), rabbit polyclonal antibody anti-TSLP Receptor (PA5-203789, or rabbit polyclonal antibody anti-IL-7Rα (1:100) (PA5-97870) (Invitrogen, Thermo Fisher Scientific, Monza, Italy).

Techniques: Expressing, Immunohistochemistry, Immunohistochemical staining, Staining, Control, Microscopy

Journal: Cells

Article Title: Human Lung-Resident Macrophages Express and Are Targets of Thymic Stromal Lymphopoietin in the Tumor Microenvironment

doi: 10.3390/cells10082012

Figure Lengend Snippet: Expression of sfTSLP, lfTSLP, TSLPR, and IL-7Rα mRNAs in peritumoral and intratumoral human lung tissues. Peritumoral and intratumoral human lung tissues (2 mg) disrupted by homogenization were lysed and 500 µL of 0.1% Triton X-100. sfTSLP and lfTSLP mRNAs were determined by quantitative RT-PCR ( A , B ). Total TSLP was evaluated in lysed peritumoral and intratumoral lung cancer tissue by ELISA ( C ). Data are mean ± SD of 5 independent experiments obtained from different patients. * p < 0.01 and § p < 0.0001 vs. peritumoral lung tissue.

Article Snippet: Paraffin-embedded sections were processed for immunohistochemistry by peroxidase anti-peroxidase method using as primary antibody rabbit polyclonal anti-TSLP antibody (1:100) (PA5-78610), rabbit polyclonal antibody anti-TSLP Receptor (PA5-203789, or rabbit polyclonal antibody anti-IL-7Rα (1:100) (PA5-97870) (Invitrogen, Thermo Fisher Scientific, Monza, Italy).

Techniques: Expressing, Homogenization, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay