Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: TSLP Interferes with Airway Tolerance by Suppressing the Generation of Antigen-specific Regulatory T cells 1

doi: 10.4049/jimmunol.1002503

Figure Lengend Snippet: A, TSLP suppressed murine iTreg differentiation in the presence of TGF-β1. Naïve CD4+CD62L+ T cells were isolated from spleens and lymph nodes, cultured with 10 μg/ml plate-bound anti-CD3 and 2 μg/ml anti-CD28 in the presence of 3 ng/ml TGF-β1 for 5 days. TSLP was used at 10 ng/ml. Data represent mean ± SEM (n = 3). B, TSLP suppressed human iTreg differentiation in the presence of TGF-β1. Naïve CD4+ T cells were isolated from PBMC and cultured in the same condition as (A). Human T cells showed considerable variability in responding to TSLP. A responder is shown on top, a non-responder on the bottom. The percentage inhibition of iTreg differentiation by TSLP from all 20 donor samples is indicated in the graph on the right. C, Purity of naïve CD4+ T cells isolated from human PBMC. Samples of PBMC and purified T cells from the same donor were stained with Blood Dendritic Cell Enumeration Kit (Miltenyi Biotec) and analyzed by flow cytometry. Data represent mean ± SEM (n = 6). D, Quantitative PCR analysis of the expression of human TSLP receptor TSLPR in activated CD4+ T cells, presented relative to naïve cells. Including of TSLP in culture (+TSLP) further enhanced TSLPR expression. Naïve CD4+ T cells were purified from PBMC and cultured in the same condition as (A). Data represent mean ± SEM (n = 6). *: p < 0.05; **: p < 0.01 One way analysis of variance.

Article Snippet: Since it is reported that human TSLP mainly exerts its effect through DCs and had little effect on CD4 + T cells ( 32 ), we examined DC contamination in our purified human naïve CD4+ T cells by Blood Dendritic Cell Enumeration Kit (Miltenyi Biotec).

Techniques: Isolation, Cell Culture, Inhibition, Purification, Staining, Flow Cytometry, Real-time Polymerase Chain Reaction, Expressing

Journal: Oncology Letters

Article Title: TSLP promotes angiogenesis of human umbilical vein endothelial cells by strengthening the crosstalk between cervical cancer cells and eosinophils

doi: 10.3892/ol.2017.7121

Figure Lengend Snippet: RhTSLP promotes VEGF secretion of HL-60E cells. Following treatment with different concentration of rhTSLP (1, 10 and 100 ng/ml) for 48 h, the secretion level of (A) VEGF and (B) FGF was determined in HL-60E cells using ELISA. The data are expressed as the mean ± standard error of the mean. *P<0.05 or ***P<0.001 vs. control, determined using one-way analysis of variance. RhTSLP, recombinant human TSLP protein; VEGF, vascular endothelial growth factor; FGF, fibroblast growth factor; NS, no statistical difference.

Article Snippet: Co-culture of HL-60E with HeLa or CasKi cells HeLa and CasKi cells (2×10 5 cells/well) were cultured with HL-60E cells (2×10 5 cells/well) in a humidified incubator with 5% CO 2 at 37°C for 48 h and subsequently incubated with or without recombinant human TSLP protein (rhTSLP; 100 ng/ml; R&D Systems, Inc., Minneapolis, MN, USA), anti-human TSLP or anti-TSLPR neutralizing antibodies (α-TSLP or α-TSLPR; 5 µg/ml; R&D Systems, Inc.) in a humidified incubator containing 5% CO 2 at 37°C for 48 h, and 1% PBS, as a negative control.

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

Journal: Oncology Letters

Article Title: TSLP promotes angiogenesis of human umbilical vein endothelial cells by strengthening the crosstalk between cervical cancer cells and eosinophils

doi: 10.3892/ol.2017.7121

Figure Lengend Snippet: TSLP-derived of cervical cancer cells upregulates the secretion of IL-8 and VEGF. HL-60E were co-cultured with HeLa or CaSki cells for 48 h and subsequently rhTSLP (100 ng/ml), α-TSLP (5 ug/ml) or α-TSLPR (5 ug/ml) was added for an additional 48 h. The secretion level of (A) IL-8, (B) VEGF and (C) FGF were determined using ELISA. The data are expressed as the mean ± standard error of the mean. *P<0.05, **P<0.01 or ***P<0.001 vs. CO of HL-60E with HeLa cells control, determined using a one-way analysis of variance; #P<0.05 or ###P<0.001 vs. CO of HL-60E with CasKi cells control, determined using a one-way analysis of variance. CO, co-culture group (HL-60E with HeLa or CaSki cells); TSLP, thymic stromal lymphopoietin; α-TSLP, anti-human TSLP neutralizing antibody; α-TSLPR, anti-human TSLP receptor neutralizing antibody.

Article Snippet: Co-culture of HL-60E with HeLa or CasKi cells HeLa and CasKi cells (2×10 5 cells/well) were cultured with HL-60E cells (2×10 5 cells/well) in a humidified incubator with 5% CO 2 at 37°C for 48 h and subsequently incubated with or without recombinant human TSLP protein (rhTSLP; 100 ng/ml; R&D Systems, Inc., Minneapolis, MN, USA), anti-human TSLP or anti-TSLPR neutralizing antibodies (α-TSLP or α-TSLPR; 5 µg/ml; R&D Systems, Inc.) in a humidified incubator containing 5% CO 2 at 37°C for 48 h, and 1% PBS, as a negative control.

Techniques: Derivative Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Co-Culture Assay

Journal: Oncology Letters

Article Title: TSLP promotes angiogenesis of human umbilical vein endothelial cells by strengthening the crosstalk between cervical cancer cells and eosinophils

doi: 10.3892/ol.2017.7121

Figure Lengend Snippet: Stimulatory effect of cervical cancer cells and EOS on angiogenesis is dependent on TSLP. Following pre-treatment with α-TSLP (5 ug/ml) or α-TSLPR (5 ug/ml) for 48 h, supernatants were selected from the co-culture system of HL-60E with (A) HeLa or (B) CaSki cells. Subsequently, HUVECs were stimulated with these supernatants and angiogenesis was analyzed using a tube formation assay. The data are expressed as the mean ± standard error of the mean. **P<0.01 vs. control, using one-way analysis of variance. EOS, eosinophils; Ctrl, control; S-H+H, supernatant of co-culture of HL-60E with HeLa cells; S-H+C, supernatant of co-culture of HL-60E with CaSki cells; EOS, eosinophils; TSLP, thymic stromal lymphopoietin; α-TSLP, anti-human TSLP neutralizing antibody; α-TSLPR, anti-human TSLP receptor neutralizing antibody; HUVECs, human umbilical vein endothelial cells.

Article Snippet: Co-culture of HL-60E with HeLa or CasKi cells HeLa and CasKi cells (2×10 5 cells/well) were cultured with HL-60E cells (2×10 5 cells/well) in a humidified incubator with 5% CO 2 at 37°C for 48 h and subsequently incubated with or without recombinant human TSLP protein (rhTSLP; 100 ng/ml; R&D Systems, Inc., Minneapolis, MN, USA), anti-human TSLP or anti-TSLPR neutralizing antibodies (α-TSLP or α-TSLPR; 5 µg/ml; R&D Systems, Inc.) in a humidified incubator containing 5% CO 2 at 37°C for 48 h, and 1% PBS, as a negative control.

Techniques: Co-Culture Assay, Tube Formation Assay

Journal: PLoS ONE

Article Title: Application of a phenotypic drug discovery strategy to identify biological and chemical starting points for inhibition of TSLP production in lung epithelial cells

doi: 10.1371/journal.pone.0189247

Figure Lengend Snippet: Production of TSLP in NHBE cells from the 2 donors used in this study.

Article Snippet: For synthesis of cDNA and qPCR, 2 qPCR reactions were performed using a TSLP (Hs01572933_m1) (FAM|L: 2900) or a GAPDH (Hs03929097_g1) (VIC_PL– 2900) TaqMan TM probe (Thermo Fisher Scientific).

Techniques:

Journal: PLoS ONE

Article Title: Application of a phenotypic drug discovery strategy to identify biological and chemical starting points for inhibition of TSLP production in lung epithelial cells

doi: 10.1371/journal.pone.0189247

Figure Lengend Snippet: Left panel shows % inhibition of TSLP production (vertical axis) for all screened compounds distributed by plates (horizontal axis). Upper horizontal blue line in the left panel indicates cut off value for hit selection (45%). Right panel is a summary of the hit selection process (DR = dose response evaluation, I = inhibition).

Article Snippet: For synthesis of cDNA and qPCR, 2 qPCR reactions were performed using a TSLP (Hs01572933_m1) (FAM|L: 2900) or a GAPDH (Hs03929097_g1) (VIC_PL– 2900) TaqMan TM probe (Thermo Fisher Scientific).

Techniques: Inhibition, Selection

Journal: PLoS ONE

Article Title: Application of a phenotypic drug discovery strategy to identify biological and chemical starting points for inhibition of TSLP production in lung epithelial cells

doi: 10.1371/journal.pone.0189247

Figure Lengend Snippet: For TSLP protein and mRNA determinations results were obtained using NHBE cells from donor #1. Results are shown as geometric mean IC 50 (μM) (95% confidence interval) or average % inhibition ± SD at the indicated concentration [μM]. Specific activities evaluated are indicated in each case and shown in bold. Activities reported in a , b , c , d and e .

Article Snippet: For synthesis of cDNA and qPCR, 2 qPCR reactions were performed using a TSLP (Hs01572933_m1) (FAM|L: 2900) or a GAPDH (Hs03929097_g1) (VIC_PL– 2900) TaqMan TM probe (Thermo Fisher Scientific).

Techniques: Inhibition, Concentration Assay

Journal: PLoS ONE

Article Title: Application of a phenotypic drug discovery strategy to identify biological and chemical starting points for inhibition of TSLP production in lung epithelial cells

doi: 10.1371/journal.pone.0189247

Figure Lengend Snippet: For TSLP protein determinations, results were obtained using NHBE cells from donor #1. Results are shown as geometric mean IC 50 (μM) (95% confidence interval) or average % inhibition ± SD at the indicated concentration [μM]. Specific activities evaluated are indicated in each case and shown in bold.

Article Snippet: For synthesis of cDNA and qPCR, 2 qPCR reactions were performed using a TSLP (Hs01572933_m1) (FAM|L: 2900) or a GAPDH (Hs03929097_g1) (VIC_PL– 2900) TaqMan TM probe (Thermo Fisher Scientific).

Techniques: Inhibition, Concentration Assay

Journal: PLoS ONE

Article Title: Application of a phenotypic drug discovery strategy to identify biological and chemical starting points for inhibition of TSLP production in lung epithelial cells

doi: 10.1371/journal.pone.0189247

Figure Lengend Snippet: Inhibition of TSLP production by mTOR inhibitors everolimus and AZD8055.

Article Snippet: For synthesis of cDNA and qPCR, 2 qPCR reactions were performed using a TSLP (Hs01572933_m1) (FAM|L: 2900) or a GAPDH (Hs03929097_g1) (VIC_PL– 2900) TaqMan TM probe (Thermo Fisher Scientific).

Techniques: Inhibition

Journal: PLoS ONE

Article Title: Application of a phenotypic drug discovery strategy to identify biological and chemical starting points for inhibition of TSLP production in lung epithelial cells

doi: 10.1371/journal.pone.0189247

Figure Lengend Snippet: Main pharmacological activities identified in compound 4.

Article Snippet: For synthesis of cDNA and qPCR, 2 qPCR reactions were performed using a TSLP (Hs01572933_m1) (FAM|L: 2900) or a GAPDH (Hs03929097_g1) (VIC_PL– 2900) TaqMan TM probe (Thermo Fisher Scientific).

Techniques: Inhibition, Activity Assay

Journal: PLoS ONE

Article Title: Application of a phenotypic drug discovery strategy to identify biological and chemical starting points for inhibition of TSLP production in lung epithelial cells

doi: 10.1371/journal.pone.0189247

Figure Lengend Snippet: Results obtained using NHBE cells from donor #1 in the TSLP assay. Results shown as geometric mean IC 50 , μM (95% confidence interval) or average % inhibition ± SD at the indicated concentration. Biochemical and cell based specific activities evaluated are indicated in each case and shown in bold.

Article Snippet: For synthesis of cDNA and qPCR, 2 qPCR reactions were performed using a TSLP (Hs01572933_m1) (FAM|L: 2900) or a GAPDH (Hs03929097_g1) (VIC_PL– 2900) TaqMan TM probe (Thermo Fisher Scientific).

Techniques: Inhibition, Concentration Assay

Journal: ERJ Open Research

Article Title: Inhibition of IL-4Rα reduces CCL26 in bronchial epithelial cells from COPD patients

doi: 10.1183/23120541.00813-2024

Figure Lengend Snippet: Interleukin (IL)-4Rα monoclonal antibody (mAb) treatment and effect on thymic stromal lymphopoietin (TSLP) and mucin 5AC (MUC5AC) gene expression in COPD epithelial cells during IL-4 or IL-13 cytokine incubation (30 ng·mL −1 ) and viral infection (multiplicity of infection 0.1). a and b) TSLP gene expression in COPD BECs 24 h after HRV1B infection and no significant changes when treated with IL-4Rα mAb and incubated with IL-4 or IL-13. MUC5AC gene expression (c and d) significantly increased after stimulation with IL-4 and IL-13, however, no significant decrease with IL-4Rα mAb treatment was observed. Results are shown as mean± sem for five individual donors (biological duplicates) of each duplicate samples in indicated gene and protein. Statistical methods used are Friedman's test and Dunn's multiple comparison test. p<0.05 was considered significant. *: p<0.05. **: p<0.01. Ab: antibody; HRV: human rhinovirus.

Article Snippet: IFN-β1 (Hs01077958_s1), TSLP (Hs00263639_m1), melanoma differentiation-associated protein 5 (MDA5) (Hs00223420_m1), retinoic acid-inducible gene I (RIG-I) (Hs00223420_m1), Toll-like receptor 3 (TLR3) (Hs01551079_g1), CCL26 (Hs00171146_m1) and mucin 5AC (MUC5AC) (Hs01365616_m1).

Techniques: Gene Expression, Incubation, Infection, Comparison

Journal: Molecules

Article Title: Lignans from Machilus thunbergii as Thymic Stromal Lymphopoietin Inhibitors

doi: 10.3390/molecules26164804

Figure Lengend Snippet: The inhibitory effects of isolated compounds 1 – 3 on thymic stromal lymphopoietin (TSLP)/TSLP receptor (TSLPR) interaction by ELISA.

Article Snippet: For the pSTAT5 assay measuring TSLP inhibitory activity, recombinant human TSLP was purchased from R&D Systems (Minneapolis, MN, USA) and BD Cytofix/Cytoperm and Alexa Fluor 647 mouse anti-STAT5 (pY694) were purchased from BD Biosciences (San Diego, CA, USA).

Techniques: Isolation, Enzyme-linked Immunosorbent Assay, Inhibition, Control

Journal: Molecules

Article Title: Lignans from Machilus thunbergii as Thymic Stromal Lymphopoietin Inhibitors

doi: 10.3390/molecules26164804

Figure Lengend Snippet: Molecular docking results of isolated compounds 1 – 3 on TSLP/TSLPR interaction.

Article Snippet: For the pSTAT5 assay measuring TSLP inhibitory activity, recombinant human TSLP was purchased from R&D Systems (Minneapolis, MN, USA) and BD Cytofix/Cytoperm and Alexa Fluor 647 mouse anti-STAT5 (pY694) were purchased from BD Biosciences (San Diego, CA, USA).

Techniques: Isolation

Journal: Molecules

Article Title: Lignans from Machilus thunbergii as Thymic Stromal Lymphopoietin Inhibitors

doi: 10.3390/molecules26164804

Figure Lengend Snippet: Molecular docking results of isolated compounds 1 – 3 against human TSLP in complex with TSLPR and interleukin-7 receptor α chain (IL-7Rα): ( A ) three-dimensional (3D) docking image of compound 1 ; ( B ) two-dimensional (2D) docking image of compound 1 ; ( C ) 3D docking image of compound 2 ; ( D ) 2D image picture of compound 2 ; ( E ) 3D docking image of compound 3 ; ( F ) 2D docking picture of compound 3 .

Article Snippet: For the pSTAT5 assay measuring TSLP inhibitory activity, recombinant human TSLP was purchased from R&D Systems (Minneapolis, MN, USA) and BD Cytofix/Cytoperm and Alexa Fluor 647 mouse anti-STAT5 (pY694) were purchased from BD Biosciences (San Diego, CA, USA).

Techniques: Isolation

Journal: Oncotarget

Article Title: Thymic stromal lymphopoietin in human pancreatic ductal adenocarcinoma: expression and prognostic significance

doi: 10.18632/oncotarget.25997

Figure Lengend Snippet: ( A ) Cytokine mRNA levels in BxPC-3, PT-45 and Capan-2 cells were assessed by real time RT-PCR and normalized to β-actin mRNA levels. Values are means (±SD) normalized gene expression. ( B ) Concentration of TSLP in CM derived from BxPC-3, PT-45, and Capan-2 cells measured by ELISA. The mean level (±SD) of cytokine detected in triplicate in CM samples is indicated.

Article Snippet: Neutralizing polyclonal antibody against TSLP (R&D Systems) or irrelevant control IgG were added to BxPC-3 cell-CM at the concentration used to neutralize TSLP biological activity (40 ng/ml) as reported above.

Techniques: Quantitative RT-PCR, Gene Expression, Concentration Assay, Derivative Assay, Enzyme-linked Immunosorbent Assay

Journal: Oncotarget

Article Title: Thymic stromal lymphopoietin in human pancreatic ductal adenocarcinoma: expression and prognostic significance

doi: 10.18632/oncotarget.25997

Figure Lengend Snippet: Incidence of OX40L expression on normal HLA-DR + DCs obtained ( A ) in the absence (control), and in the presence of 20% BxPC-3 or Capan-2 cell-CM. Values are means ± SE. P -values obtained by One Way ANOVA followed by Bonferroni t -Test, or ( B ) in the presence of BxPC-3 cell-CM, pre-treated with a neutralizing anti-TSLP polyclonal antibody. Values are means ± SE. P -values obtained by the Paired t -Test.

Article Snippet: Neutralizing polyclonal antibody against TSLP (R&D Systems) or irrelevant control IgG were added to BxPC-3 cell-CM at the concentration used to neutralize TSLP biological activity (40 ng/ml) as reported above.

Techniques: Expressing, Control

Journal: Oncotarget

Article Title: Thymic stromal lymphopoietin in human pancreatic ductal adenocarcinoma: expression and prognostic significance

doi: 10.18632/oncotarget.25997

Figure Lengend Snippet: Basal circulating TSLP levels in PDAC patients ( n = 82) and healthy donors ( n = 41) as determined by ELISA. P -values obtained by the Mann-Whitney Rank Sum Test. Median, 10th, 25th, 75th, and 90th percentiles are presented as vertical boxes with error bars. Dots indicate outliers.

Article Snippet: Neutralizing polyclonal antibody against TSLP (R&D Systems) or irrelevant control IgG were added to BxPC-3 cell-CM at the concentration used to neutralize TSLP biological activity (40 ng/ml) as reported above.

Techniques: Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

Journal: Oncotarget

Article Title: Thymic stromal lymphopoietin in human pancreatic ductal adenocarcinoma: expression and prognostic significance

doi: 10.18632/oncotarget.25997

Figure Lengend Snippet: Plasma levels of TSLP pre- and post-treatment

Article Snippet: Neutralizing polyclonal antibody against TSLP (R&D Systems) or irrelevant control IgG were added to BxPC-3 cell-CM at the concentration used to neutralize TSLP biological activity (40 ng/ml) as reported above.

Techniques: Clinical Proteomics