Journal: Cancer Research

Article Title: Specific Recruitment of CC Chemokine Receptor 4–Positive Regulatory T Cells in Hodgkin Lymphoma Fosters Immune Privilege

doi: 10.1158/0008-5472.can-06-0261

Figure Lengend Snippet: Figure 1. Chemotactic activity of the supernatants of HL cell lines. The chemotactic responses of human PBL induced by the supernatants of HL and ALCL cell lines, rh-TARC/CCL17 and rh-MDC/CCL22. The number of cells migrated (count/AL) into the lower well containing control medium is set as ‘‘one’’ and the relative numbers of cells of each subset migrated into the lower well containing the chemoattractants are presented. Horizontal dotted line, base unit. The supernatants of the HL cell lines did not show any chemotactic activity for whole lymphocytes (A) and CD8+

Article Snippet: Recombinant human (rh)-TARC/CCL17 (R&D Systems) and/or rh-MDC/ CCL22 (R&D Systems) were used as control chemoattractants.

Techniques: Activity Assay, Control

Journal: Cancer Research

Article Title: Specific Recruitment of CC Chemokine Receptor 4–Positive Regulatory T Cells in Hodgkin Lymphoma Fosters Immune Privilege

doi: 10.1158/0008-5472.can-06-0261

Figure Lengend Snippet: Figure 5. The attribution of the chemotactic activity of the supernatant of the HL cell line to TARC/CCL17 and MDC/CCL22 and KM2760-induced inhibition of the migration of CD4+CD25+ T cells. A to C, percentages were determined of CD4+ cells in PBMC, CD4+CCR4+ cells in PBMC, and CD4+CCR4+ cells in CD4+ cells, obtained from 15 healthy adult volunteers, incubated with KM2760 or Rituximab or without any antibody. Each closed circle represents the percentage of CD4+ cells in each PBMC, CD4+CCR4+ cells in each PBMC, and CD4+CCR4+ cells in each CD4+ cell population. Points, mean; bars, SD. *, P < 0.05, significant differences between each group (n.s., not significant). A 12-hour treatment with KM2760 significantly decreased the proportions of CD4+ cells in PBMC, CD4+CCR4+ cells in PBMC, and CD4+CCR4+ cells in CD4+ cells. D, the source of the chemotactic activity in the supernatants of the HL cell lines was examined by a chemotaxis assay of human PBL using the neutralizing antibodies antihuman TARC/CCL17 and antihuman MDC/CCL22 mAbs. Antihuman TARC/CCL17 alone or antihuman MDC/CCL22 mAb alone did not completely inhibit the migration of the CD4+CD25+ T cells induced by the supernatant of one of the HL cell lines, HDLM-2, which produces both TARC/CCL17 and MDC/CCL22. In contrast, the presence of both antibodies together caused an almost complete inhibition of the CD4+CD25+

Article Snippet: Recombinant human (rh)-TARC/CCL17 (R&D Systems) and/or rh-MDC/ CCL22 (R&D Systems) were used as control chemoattractants.

Techniques: Activity Assay, Inhibition, Migration, Incubation, Chemotaxis Assay

Journal: bioRxiv

Article Title: Macrophages foster adaptive anti-tumor immunity by ZEB1-dependent cytotoxic T cell chemoattraction

doi: 10.1101/2024.02.26.582102

Figure Lengend Snippet: A . Differentially expressed genes in unstimulated or LPS or IL-4 stimulated LysM ΔZeb1 compared to LysM Ctrl BMDMs as measured by a customized RT2 array and depicted in log 2 fold change of expression. nd marks non-detectable mRNA levels. All transcripts were normalized to Gapdh (n=3). B . Relative mRNA expression of Ccl2 and Ccl22 in LysM Ctrl and LysM ΔZeb1 BMDMs (n=3; means ±SD; 2-way ANOVA). C . Comparison of transcript and secretome alterations of Ccl2 and Ccl22 in LysM ΔZeb1 compared to LysM Ctrl BMDMs (n≥5; means ±SD).

Article Snippet: Final concentrations of cytokines were 1.93 ng/ ml CCL2 (R&D Systems, 479-JE-050) and 0.18 ng/ ml CCL22 (R&D Systems, 439-MD-025) and of antibodies 5 μg/ml IgG (Diagenode, C15410206), anti-CCL2 (Novus Biologicals, NBP1-07035SS) or anti-CCL22 (abcam, ab124768), respectively.

Techniques: Expressing, Comparison

Journal: bioRxiv

Article Title: Macrophages foster adaptive anti-tumor immunity by ZEB1-dependent cytotoxic T cell chemoattraction

doi: 10.1101/2024.02.26.582102

Figure Lengend Snippet: A-B . Venn diagrams of differentially expressed genes (DEGs) of LysM Ctrl (blue) and LysM ΔZeb1 BMDMs (red) after stimulation with LPS (A) or IL-4 (B) compared to unstimulated in total (left) and divided in up-/downregulated DEGs (right). C . GO term enrichment analysis for DEGs (FDR<0.05) uniquely up- or downregulated by LysM ΔZeb1 BMDMs after LPS stimulation. D . Log 2 fold change of expression of selected trafficking genes after LPS stimulation. X marks no significant deregulation. E . Representative images and quantification of OPP incorporation of LysM Ctrl and LysM ΔZeb1 BMDMs with 0h, 4h and 16h LPS pre-stimulation (n=3; means ±SD; 2-way ANOVA). F-G . Representative arrays of intracellular cytokines of LysM Ctrl and LysM ΔZeb1 BMDMs with LPS stimulation or additional Brefeldin A and Monensin treatment (F) and quantification of intracellular CCL2 and CCL22 after LPS, Brefeldin A and Monensin treatment (G; n≥2).

Article Snippet: Final concentrations of cytokines were 1.93 ng/ ml CCL2 (R&D Systems, 479-JE-050) and 0.18 ng/ ml CCL22 (R&D Systems, 439-MD-025) and of antibodies 5 μg/ml IgG (Diagenode, C15410206), anti-CCL2 (Novus Biologicals, NBP1-07035SS) or anti-CCL22 (abcam, ab124768), respectively.

Techniques: Expressing

Journal: bioRxiv

Article Title: Macrophages foster adaptive anti-tumor immunity by ZEB1-dependent cytotoxic T cell chemoattraction

doi: 10.1101/2024.02.26.582102

Figure Lengend Snippet: A . Confluence of KPC cells alone or co-cultured with LysM Ctrl or LysM ΔZeb1 BMDMs (n=3). B . Representative images at t=28h and quantification over time of KPC cell invasion into a scratch wound without or with co-culture of LysM Ctrl or LysM ΔZeb1 BMDMs (n=3). C . Confluence of KPC cells alone or with LysM Ctrl or LysM ΔZeb1 BMDM conditioned medium (CM) (n=2 KPC CM, n=3 LysM Ctrl and LysM ΔZeb1 CM). D . Transwell migration assay of CD8+ T cells alone or towards LysM Ctrl or LysM ΔZeb1 BMDMs in absence or presence of recombinant CCL2 and CCL22 (left panel, n>3) or absence or presence of anti-CCL2 and anti-CCL22 antibodies (right panel, n=3). Means ±SD; *:p<0.05; **:p<0.01; ns: not significant; 2-way ANOVA.

Article Snippet: Final concentrations of cytokines were 1.93 ng/ ml CCL2 (R&D Systems, 479-JE-050) and 0.18 ng/ ml CCL22 (R&D Systems, 439-MD-025) and of antibodies 5 μg/ml IgG (Diagenode, C15410206), anti-CCL2 (Novus Biologicals, NBP1-07035SS) or anti-CCL22 (abcam, ab124768), respectively.

Techniques: Cell Culture, Co-Culture Assay, Transwell Migration Assay, Recombinant