anti ccl5 neutralizing antibodies  (R&D Systems)

Journal: Signal Transduction and Targeted Therapy

Article Title: Employing epigenetic protein degradation techniques to block CCL5-mediated photodynamic therapy via a programmed delivery platform

doi: 10.1038/s41392-025-02542-y

Figure Lengend Snippet: Mechanistic study of the antitumor effects of ARV/Ce6@RDP micelles. a Venn diagrams of the intersection between upregulated genes in Ce6@RDP (+) vs RDP and downregulated genes in ARV/Ce6@RDP (+) vs Ce6@RDP (+) for 4T1 and B16F10 cells based on RNA-seq analysis. b Venn diagrams of the intersection between upregulated genes in Ce6@RDP (+) vs RDP and upregulated genes in ARV/Ce6@RDP (+) vs Ce6@RDP (+) for 4T1 and B16F10 cells. c Heatmap of the intersection genes from ( a ) and ( b ) expressed in 4T1 and B16F10 cells receiving different treatments. d Quantification of the CCL5 level in the supernatants of treated 4T1 and B16F10 cells ( n = 3 per group, two-tailed unpaired Student’s t -test). Cell viability ( e ) and apoptosis ( f ) analysis of Ccl5 -knockdown 4T1 and B16F10 cells with and without treatment with PDT ( n = 3 per group, two-tailed unpaired Student’s t -test). g – n Effects of Ccl5 knockdown on PDT efficacy in vivo. Tumor growth curves ( g , n = 6 per group, two-way ANOVA with Tukey’s multiple comparisons test), post-treatment tumor photographs ( h ), and tumor weight analysis ( i , n = 6 per group, two-tailed unpaired Student’s t -test) of Ccl5 -knockdown 4T1 cells and control cell xenografts treated with or without PDT; Tumor growth curves ( j , n = 5 per group, two-way ANOVA with Tukey’s multiple comparisons test), post-treatment tumor photographs ( k ) and tumor weight analysis ( l , n = 5 per group, two-tailed unpaired Student’s t -test) of Ccl5 -knockdown B16F10 cells and control cell xenografts treated with or without PDT; Expression levels of the CCL5 protein ( m ) and proteins ( n ) related to proliferation and apoptosis in tumor tissues collected from Ccl5 -knockdown 4T1 and B16F10 xenografts after treatment with or without PDT. o Flow cytometric analyses of M2 polarization in BMDMs after treatment with culture medium from Ccl5 -knockdown 4T1 cells or control cells ( n = 3 per group, two-tailed unpaired Student’s t -test). p M2 polarization analysis of BMDMs treated with culture medium from Ccl5 -overexpressing 4T1 cells and control cells ( n = 3 per group, two-tailed unpaired Student’s t -test). q Mrc1 , Arg1 , Irf4 , Ym1 , and Cd274 gene expression in BMDMs incubated with culture medium from different 4T1 cells determined by qPCR ( n = 3 per group, two-tailed unpaired Student’s t -test). r Recruitment of M2 macrophages by Ccl5 -knockdown or Ccl5 -overexpressing 4T1 cells ( n = 3 per group; two-tailed unpaired Student’s t -test; scale bar: 50 µm). s CFSE staining analysis of 4T1 cell proliferation in BMDMs subjected to different treatments ( n = 3 per group, two-tailed unpaired Student’s t -test). t Effects of nanomedicines on Ccl5 promoter activity detected by dual-luciferase assay ( n = 3 per group, two-tailed unpaired Student’s t -test). u ChIP‒qPCR analysis of BRD4 protein binding to the Ccl5 promoter in B16F10 cells ( n = 3 per group; two-way ANOVA with Sidak’s multiple comparisons test). v Schematic diagram of ARV-825-mediated inhibition of Ccl5 gene transcription to enhance PDT. The figure was created with Figdraw.com. The data are presented as the means ± SDs for in vitro experiments and means ± SEMs for in vivo experiments

Article Snippet: For IHC, the slides were subsequently separately stained with primary antibodies against Ki67 (# GB111141 , Servicebio), CD31 (# GB113151 , Servicebio), PDL1 (#ab213480, Abcam), CCL5 (SC-365826, Santa Cruz), CD206 (# GB113497 , Servicebio), and Foxp3 (# GB112325 , Servicebio), followed by incubation with an HRP-conjugated secondary antibody.

Techniques: RNA Sequencing, Two Tailed Test, Knockdown, In Vivo, Control, Expressing, Gene Expression, Incubation, Staining, Activity Assay, Luciferase, Protein Binding, Inhibition, In Vitro

Journal: Advanced Science

Article Title: Loss of SOCS1 in Donor T Cells Exacerbates Intestinal GVHD by Driving a Chemokine‐Dependent Pro‐Inflammatory Immune Microenvironment

doi: 10.1002/advs.202513735

Figure Lengend Snippet: Socs1 cKO Drives Monocyte Polarization Toward M1 Macrophages in Mice. (A) UMAP plot of annotated subsets extracted from MDM cells based on scRNA‐seq data in Figure , colored by clusters, and integrated with RNA velocity analysis to illustrate predicted future transcriptional states and developmental trajectories of the subsets. UMAP plots of feature genes are displayed around the main plot. (B) UMAP plots showing the distribution of MDM cells in WT and cKO groups, and the visualization of M1 and M2 signature scores. (C) Differential expression genes (DEGs) analysis of Monocyte‐like cells between WT and cKO groups. Red indicated upregulation in the cKO group, and blue indicated downregulation in the cKO group. (D) KEGG pathway analysis based on DEGs in Figure . (E) Representative flow cytometry plots and frequencies of F4/80 low CD11b hi Macrophage, Monocyte‐like cells, and iNOS + Macrophage in the spleen of WT and cKO mice (n = 5 per group). (F) Quantification of macrophage subpopulations in WT and cKO mice. Bar plots show the frequency (% of total single cells) of the indicated macrophage populations in the spleen as determined by flow cytometry. Populations shown include total macrophages, monocyte‐like, iNOS + , Ly6C + , CD80 + , CD80 + , CD86 + , CD206 + , MHC class I + and MHC class II + macrophages. (G) iTALK‐identified and visualized ligand‐receptor interaction signals between Monocyte‐like subsets and CD8 + Teff subsets, illustrating differential intercellular communication patterns in the cKO group compared to the WT group. Legend is shown in Figure S3G. (H) Transwell migration assay showing the migration rate of monocytes from WT mice toward CD8 + T cells derived from WT or cKO mice after 2 h (left) and 12 h (right) (n = 5). (I) Transwell migration assay showing the 12‐h migration rate of monocytes from WT mice toward CD8 + T cells derived from WT or cKO mice, either with or without CCL5 activation (rmCCL5, left panel), and with CCL5 neutralization using an anti‐CCL5 antibody (right panel) (n=4). (J) Experimental schematic showing the co‐culture system of BMDMs and CD8 + T cells. (K) Barplot and flow cytometry histogram showing expression of activation markers (CD80, CD86) and MHC Class II on macrophages following co‐culture with T cells from two groups. Data represent one experiment out of two independent experiments. P values were determined using an unpaired two‐tailed Student's t‐test (F, H‐I, K). Data represent mean ± SD (F, H‐I, K). ∗ ∗P <.05, ∗∗P <.01 and ∗∗∗∗P <.0001.

Article Snippet: Recipient mice were treated with anti‐CCL5 neutralizing antibodies (10 μg/mouse; R&D Systems, AF478) or isotype control antibodies.

Techniques: Quantitative Proteomics, Flow Cytometry, Transwell Migration Assay, Migration, Derivative Assay, Activation Assay, Neutralization, Co-Culture Assay, Expressing, Two Tailed Test

Journal: Advanced Science

Article Title: Loss of SOCS1 in Donor T Cells Exacerbates Intestinal GVHD by Driving a Chemokine‐Dependent Pro‐Inflammatory Immune Microenvironment

doi: 10.1002/advs.202513735

Figure Lengend Snippet: Inhibiting JAK/STAT signaling or CCL5 alleviates intestinal GVHD in Socs1 ‐deficient models. (A) Experimental schematic. Recipient mice were transplanted with 5 × 10 6 T‐cell depleted bone marrow (TCD‐BM) cells from WT mice and 1 × 10 6 splenic T cells from SOCS1‐cKO mice. Mice were treated with either control IgG or anti‐CCL5 antibody on Day 3, Day5, Day7 and Day9 post‐transplantation. (B–D) Survival curve (B), GVHD score (C), and body weight changes (D) of recipient mice treated with IgG or anti‐CCL5. Data are pooled from two independent experiments with a total of 8 mice per group. The median survival was 26 days in the IgG group versus 33.5 days in the anti‐CCL5 group. (E) Representative images of HE stained intestinal crypt architecture on Day 7 and Day 14 post‐transplantation in the IgG group and the anti‐CCL5 group. Scale bar, 200 µm. (F) Quantitative analysis of crypt depth (µm) and crypt area (µm 2 ) on Day 7 and Day 14 post‐transplantation in the IgG group and anti‐CCL5 group. (G) Representative multiplex immunofluorescence images of the small intestine from mice treated with control IgG or anti‐CCL5 antibody, analyzed on Day 7 and 14 post‐transplantation. Sections were stained for CD8 (yellow), F4/80 (green), and DAPI (blue). Scale bar, 50 µm. (H) Quantification of the abundance of F4/80 + cells in the small intestine at the indicated time points. (I) Experimental schematic. BALB/c recipient mice were transplanted with 5 × 10 6 TCD‐BM cells from WT mice and 1×10 6 splenic T cells from cKO mice and treated with control vehicle, JAK/STAT inhibitor Ruxolitinib (30 mg/kg, i.g., BID), CCR5 antagonist Maraviroc (30 mg/kg, i.p., QD) or the combination from Day 1 to Day 20. Survival was monitored daily. Body weight and GVHD score were assessed every five days. (J‐L) Survival curve (J), GVHD score (K), and body weight changes (L) of the recipients in four groups. Each group included 12‐13 mice from two independent experiments. The median survival time in the control group was 11 days. At the observation endpoint (Day 70), 8/12 mice in the Ruxolitinib group, 1/12 mice in the Maraviroc group, and 9/13 mice in the Combination group remained alive. (M) Proportions of CD8 + T cells and monocytes in PB of recipients on Day 7, Day 14, and Day 21 (top, n = 3‐5 per group); and proportion of CD8+ T cells and Ly6Chi macrophage in the intestinal IEL of recipients on Day 7 (bottom, n = 3 per group) among indicated groups. (N) Representative images of HE stained intestinal crypt architecture on Day 7 and Day 14 post‐transplantation in the control, Ruxolitinib, and Maraviroc treatment groups. Scale bar, 200 µm. (O) Quantitative analysis of crypt depth (µm) and crypt area (µm 2 ) on Day 7 and Day 14 post‐transplantation in the control, Ruxolitinib, and Maraviroc treatment groups. Data represent two independent experiments. P values were determined using chi‐squared test (B, J) or unpaired two‐tailed Student's t‐test (F, M, O). Weight loss and GVHD score (C‐D, K‐L) were analyzed by one‐way ANOVA. Data represent mean ± SEM (C‐D, F, K‐L, M, O). ∗ p <.05, ∗∗ p <.01, ∗∗∗ p <.001 and ∗∗∗∗ p <.0001.

Article Snippet: Recipient mice were treated with anti‐CCL5 neutralizing antibodies (10 μg/mouse; R&D Systems, AF478) or isotype control antibodies.

Techniques: Control, Transplantation Assay, Staining, Multiplex Assay, Immunofluorescence, Two Tailed Test