anti cc chemokine ligand 17 ccl17 antibody  (R&D Systems)

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: Nature Cardiovascular Research

Article Title: Identification of a non-canonical chemokine-receptor pathway suppressing regulatory T cells to drive atherosclerosis

doi: 10.1038/s44161-023-00413-9

Figure Lengend Snippet: Fig. 5 | CCL17–CCR8–CCL3 axis critically interferes with Treg differentiation. a, Schematic of co-culture experiment, where isolated splenic CD4+CD62L+ T cells from Ccr8WTApoe−/− or Ccr8KOApoe−/− mice were combined with sorted CD11c+MHCII+ cDCs from LN of CCR8WTApoe−/− mice and cultured for 3 d in the absence or presence of recombinant murine CCL17 (100 ng ml−1). b, CCL3 concentrations were measured in cell supernatants by ELISA (number of independent experiments per bar from left to right, n = 7, 9, 6, 9). c, CD4+CD25+Foxp3+ Treg cells were quantified using flow cytometry analysis (number of independent experiments per bar from left to right: n = 10, 12, 10 and 13). d, Scheme of co-culture experiment, where isolated splenic CD4+CD62L+ T cells from Ccr8WTApoe−/− or Ccr8KOApoe−/−mice were combined with sorted CD11c+MHCII+ cDCs from LN of Apoe−/− or Apoe−/−Ccl17e/e mice and cultured for 3 d. e, CCL3 concentrations in the supernatant were determined by ELISA

Article Snippet: Proximity ligation was carried out using the Duolink In Situ Red kit goat/rabbit (Sigma-Aldrich) on PFA-fixated mouse DCs cultured on collagen-coated coverslips that were pre-incubated with recombinant mouse CCL1 (Peprotech) and CCL17 (BioLegend) using primary polyclonal antibodies to mouse CCL17 (R&D systems), mouse CCL1 (Acris), mouse CCR4 (Thermo Scientific), mouse CCR5 (Santa Cruz Biotechnology) and mouse CCR8 (Abcam) according to the manufacturer’s instructions.

Techniques: Co-Culture Assay, Isolation, Cell Culture, Recombinant, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Nature Cardiovascular Research

Article Title: Identification of a non-canonical chemokine-receptor pathway suppressing regulatory T cells to drive atherosclerosis

doi: 10.1038/s44161-023-00413-9

Figure Lengend Snippet: Fig. 7 | CCL17-dependent CCL3 release controls Treg differentiation via CCR1. a, Experimental scheme wherein CD4+CD62L+ T cells isolated from spleens of Apoe−/−, Apoe−/−Ccr1−/− or Apoe−/−Ccr5−/− mice were cultured for 3 d under Treg-polarizing conditions (TGFβ at 100 ng ml−1) in the presence or absence of recombinant mouse CCL3 (100 ng ml−1). b, Quantification of CD45+CD4+CD25+FoxP3+ Treg cells (number of independent experiments per bar from left to right, n = 7, 7, 7, 7, 7, 7, 6, 6 and 6) using flow cytometry. c, Scheme of co-culture experiment wherein CD4+CD62L+ T cells isolated from spleens of Apoe−/−, Apoe−/−Ccr1−/− or Apoe−/−Ccr5−/− mice were combined with sorted CD45+CD11c+MHCII+eGFP+ cDCs from LNs of Apoe−/−Ccl17wt/e or Apoe−/−Ccl17e/e mice and cultured for 3 d. d, Quantification of CD45+CD4+CD25+FoxP3+ Treg cells (number of independent experiments per bar from left to right, n = 5, 5, 5, 4, 5 and 3) using flow cytometry. e, Experimental scheme of Apoe−/− or Apoe−/−Ccr1−/−

Article Snippet: Proximity ligation was carried out using the Duolink In Situ Red kit goat/rabbit (Sigma-Aldrich) on PFA-fixated mouse DCs cultured on collagen-coated coverslips that were pre-incubated with recombinant mouse CCL1 (Peprotech) and CCL17 (BioLegend) using primary polyclonal antibodies to mouse CCL17 (R&D systems), mouse CCL1 (Acris), mouse CCR4 (Thermo Scientific), mouse CCR5 (Santa Cruz Biotechnology) and mouse CCR8 (Abcam) according to the manufacturer’s instructions.

Techniques: Isolation, Cell Culture, Recombinant, Flow Cytometry, Co-Culture Assay