Journal: Experimental & Molecular Medicine

Article Title: H3K18 lactylation in cancer-associated fibroblasts drives malignant pleural effusion progression via TNFR2 + T reg recruitment

doi: 10.1038/s12276-025-01557-3

Figure Lengend Snippet: a KEGG pathway enrichment of DEGs between FACS-sorted TNFR2 + and TNFR2 − CD4 + T cells from MPE ( n = 3). b , c Volcano plot ( b ) and heatmap ( c ) of DEGs related to the chemokine signaling pathway. d , e Flow cytometry histograms ( d ) and comparisons of chemokine receptor expression (CXCR6, CCR4 and CCR6) ( e ) on TNFR2 + T reg cells and TNFR2 − T reg cells ( n = 13). f Schematic of Transwell chemotaxis assay testing TNFR2 − T reg chemotaxis toward MPE supernatant (by Figdraw). g , h Transwell chemotaxis assay comparing TNFR2 + T reg frequencies between freshly isolated cells and cells that migrated toward MPE supernatant after 4 h incubation. i Concentrations of CXCL16, CCL17, CCL22 and CCL20 in MPE and PB were quantified using ELISA ( n = 16). j Schematic of chemotaxis assay to investigate the chemotatic axis to attract TNFR2 + T reg cells in MPE. k , l Flow cytometry histograms and comparisons of chemotaxis of TNFR2 + T reg cells in response to MPE in the presence of anti-CXCL16, anti-CCL17, anti-CCL22 or anti-CCL20 mAbs. Data shown in d , e , g – i , k and l are representative of at least three independent experiments (mean ± s.d.). Statistical analysis was performed using paired two-tailed Student’s t -test ( e and h ), Wilcoxon test ( i ) or one-way ANOVA ( l ). * P < 0.05, ** P < 0.01, **** P < 0.0001. ns not significant, mAbs monoclonal antibodies.

Article Snippet: In specific experimental conditions, the following reagents were added to the lower chamber: anti-CCL17 antibody (50 ng/ml, #MAB364-SP, R&D Systems), anti-CCL20 antibody (50 ng/ml, #MAB360-SP, R&D Systems), anti-CCL22 antibody (50 ng/ml, #MAB336-SP, R&D Systems), anti-CXCL16 antibody (50 ng/ml, #MAB976, R&D Systems) or recombinant human CXCL16 (100 ng/ml, #976-CX-025/CF, R&D Systems).

Techniques: Flow Cytometry, Expressing, Chemotaxis Assay, Isolation, Incubation, Enzyme-linked Immunosorbent Assay, Two Tailed Test, Bioprocessing

Journal: Frontiers in endocrinology

Article Title: CCL17 and CCL22 chemokines are upregulated in human obesity and play a role in vascular dysfunction.

doi: 10.3389/fendo.2023.1154158

Figure Lengend Snippet: FIGURE 1 Circulating CCL17 and CCL22 chemokine levels in morbidly obese patients and age-matched controls. (A) CCL17 and (B) CCL22 levels were measured in plasma samples from obese patients (n = 60) and controls (n = 20). Scatter dot plots showing median with interquartile range. Comparison between groups were made by Mann Whitney test. Spearman test shows a positive correlation between CCL17 and CCL22 with HOMA- IR Index (C, D) and BMI (E, F) (n = 20 control subjects and n = 60 morbidly obese patients).

Article Snippet: Antigen was unmasked with proteinase K (cat#S3020, Dako, Santa Clara, CA) and blocked with 15% horse serum for 1 h. Samples were incubated with the following primary antibodies overnight at 4°C: mouse anti-human CCL17 (1:50, cat#DY364-05, R&D Systems), mouse anti-human CCL22 (1:50, cat#DY336, R&D Systems), goat anti-human CCR4 (1:100, ab1669; Abcam, Cambridge, UK), rabbit anti-human CD3 (1:100, cat#C7930, Sigma-Aldrich, St. Louis, MO), rabbit polyclonal anti-human CD31 (1:50, cat#ab32457, Abcam) and rat anti-human Mac-3 (1:100, cat#sc19991, Santa Cruz Biotechnology, Dallas, TX).

Techniques: Clinical Proteomics, Comparison, MANN-WHITNEY, Control

Journal: Frontiers in endocrinology

Article Title: CCL17 and CCL22 chemokines are upregulated in human obesity and play a role in vascular dysfunction.

doi: 10.3389/fendo.2023.1154158

Figure Lengend Snippet: FIGURE 2 Expression of CCL17 and CCL22 is increased in VCAT from morbidly obese patients. Relative quantification of mRNA levels for (A) CCL17 and (B) CCL22. Comparisons between groups were made by Wilcoxon matched-pair signed-rank test. Values are expressed as mean ± SEM (n = 33). (C) CCL17 and (D) CCL22 chemokine release into conditioned media was determined after 48 h of SCAT and VCAT explant culture. Chemokine secretion is expressed as pg/ml in the supernatant. Values are expressed as mean ± SEM (n = 22). Comparison between groups were made by Mann Whitney test. (E) Immunofluorescence representative images showing colocalization of CCL17 with CD3 (lymphocytes), CD31 (endothelial cells) and Mac-3 (macrophages); or CCL22 with CD3, CD31, Mac-3 in VCAT. Immunoreactivity was visualized using Alexa Fluor 594 (CCL17 and CCL22, red) and Alexa Fluor 488 (CD31, CD3, Mac-3, green) secondary antibodies. Nuclei were stained with Hoechst (blue). Scale bar, 20 mm. Nuclei were stained with Hoechst (blue).

Article Snippet: Antigen was unmasked with proteinase K (cat#S3020, Dako, Santa Clara, CA) and blocked with 15% horse serum for 1 h. Samples were incubated with the following primary antibodies overnight at 4°C: mouse anti-human CCL17 (1:50, cat#DY364-05, R&D Systems), mouse anti-human CCL22 (1:50, cat#DY336, R&D Systems), goat anti-human CCR4 (1:100, ab1669; Abcam, Cambridge, UK), rabbit anti-human CD3 (1:100, cat#C7930, Sigma-Aldrich, St. Louis, MO), rabbit polyclonal anti-human CD31 (1:50, cat#ab32457, Abcam) and rat anti-human Mac-3 (1:100, cat#sc19991, Santa Cruz Biotechnology, Dallas, TX).

Techniques: Expressing, Comparison, MANN-WHITNEY, Staining

Journal: Frontiers in immunology

Article Title: A novel monoclonal antibody against human thymic stromal lymphopoietin for the treatment of TSLP-mediated diseases.

doi: 10.3389/fimmu.2024.1442588

Figure Lengend Snippet: FIGURE 2 Neutralization of TSLP activity by TAVO101 and tezepelumab in functional potency assays. (A). Neutralization of human TSLP-driven STAT5 reporter gene activation by TAVO101 and tezepelumab. Increasing amounts of TAVO101 or tezepelumab along with 3 ng/mL recombinant human TSLP were applied to HEK293T cells transfected with human TSLP receptor complex and a STAT5-responsive luciferase reporter gene and reporter gene expression was quantitated. The percentages of TSLP activity normalized to the maximum activity driven by 3 ng/mL human TSLP were plotted against the concentrations of testing antibodies (Data expressed as mean ± SEM, n=3). (B). Neutralization of human TSLP-driven proliferation of BaF3 cells transfected with human TSLP receptor complex by TAVO101. Increasing amounts of TAVO101 along with 0.5 ng/mL recombinant human TSLP were applied to the transfected cells and cell proliferation was quantitated. Luminescence signals reflecting cell proliferation were plotted against the concentrations of TAVO101 (Data expressed as mean ± SEM, n=3). (C). Neutralization of TSLP-driven CCL17 release from activated dendritic cells by TAVO101. CD1c+ blood dendritic cells isolated from the PBMC of two healthy donors were treated with 15 ng/mL human TSLP with or without 1 mg/mL TAVO101. The CCL17 releases from the activated dendritic cells were quantitated. The CCL17 levels were plotted against the testing antibodies in the bar graphs as shown. (D). Neutralization of TSLP-driven proliferation of activated human CD4+ T cells. Human CD4+ T cells isolated from the PBMC of two healthy donors were labelled by Cell Proliferation Dye eFluor 450, activated by plate bound anti-CD3 antibody and treated with 50 ng/mL human TSLP with or without null control antibody, tezepelumab or TAVO101 at the indicated concentrations. The fraction of proliferated human CD4+ T cells were quantitated by flow cytometry as cells stained with diluted dyes. The fold of T cell proliferation over that without TSLP treatment were plotted against the testing antibodies in the bar graphs as shown. (Data expressed as mean ± SEM, n=2).

Article Snippet: The cell supernatants were collected and the TSLP-driven release of CCL17 protein was quantitated using the human CCL17/TARC kit (R&D Systems, Minneapolis, MN).

Techniques: Neutralization, Activity Assay, Functional Assay, Activation Assay, Recombinant, Transfection, Luciferase, Gene Expression, Isolation, Control, Cytometry, Staining

Journal: Frontiers in immunology

Article Title: A novel monoclonal antibody against human thymic stromal lymphopoietin for the treatment of TSLP-mediated diseases.

doi: 10.3389/fimmu.2024.1442588

Figure Lengend Snippet: FIGURE 3 Efficacy of TAVO101 in TSLP/OVA-induced asthma model using hTSLP/hTSLPR humanized mice. (A). Dosing regimen and animal grouping in the asthmatic model. Four mice were enrolled in the G1 group while eight mice were enrolled in each of the G2 to G6 groups. (B). The concentration of mouse total serum IgE and lung tissue CCL17 and IL-13. (C). Cell counts of mouse CD45+ leukocytes and eosinophils and the associated percentage of eosinophils in CD45+ leukocytes in BALF of asthmatic mice. (D). Scores of inflammatory cell infiltration and eosinophil infiltration by H&E staining and the positive area proportion of Goblet cells and mucus of asthmatic lungs. Note: Data was represented by mean ± SEM and analyzed by One- way ANOVA with Dunnett’s multiple comparisons test. Comparison between each experimental group and G2 group. (*p<0.05, **p<0.01, ***p<0.001, ****p<0.0001).

Article Snippet: The cell supernatants were collected and the TSLP-driven release of CCL17 protein was quantitated using the human CCL17/TARC kit (R&D Systems, Minneapolis, MN).

Techniques: Concentration Assay, Staining, Comparison

Journal: medRxiv

Article Title: NOTCH1 fusion genes in pediatric T-cell lymphoblastic lymphoma hallmark a common high-risk subgroup with blood TARC levels as possible diagnostic biomarker

doi: 10.1101/2024.01.31.24301517

Figure Lengend Snippet: A) TARC levels in pg/ml per patient, showing highly elevated TARC in blood of NOTCH1 -rearranged T-LBL patients but none of the other patients. 10000 pg/ml is the maximum measurable TARC level with used assay. Orange line in figure A, B and C represents maximum normal TARC level (1300 pg/ml) Patients that had a relapse are indicated with an asterisk. Patient TLBL049 developed a therapy-related AML (double asterisk). B) For three NOTCH1 -rearranged patients, blood TARC levels could be determined for a time point of remission after diagnosis, revealing normalized TARC levels in all three cases. C) For three NOTCH1 -rearranged patients blood TARC levels were determined at time point of relapse and remission after relapse (second remission), revealing increased levels in two relapses that again normalized in second remission. D) Staining for TARC using anti-CCL17 antibody for four NOTCH1 -rearranged patients showing that T-LBL cells do not express TARC.

Article Snippet: Immunohistochemistry staining for TARC was performed on the BOND-III fully automated staining system (Leica, IL, USA) using CCL17 rabbit polyclonal antibody (ProteinTech group, Chicago IL, USA).

Techniques: Staining