Journal: Cell Communication and Signaling : CCS

Article Title: Blocking CCR1 + macrophages overcomes resistance to immune checkpoint inhibitors in melanoma

doi: 10.1186/s12964-025-02444-0

Figure Lengend Snippet: CCR1 is mainly expressed in macrophages. A Differential signaling ligand-receptor pairs between immune cells in R and NR groups. Dot sizes represent P -values (only P <0.05 are shown). B Violin plot showing CD163, CCR1, and CCR5 expression in different immune cells. C Spatial feature plots of gene expression of CCR1 and CD68 in melanoma tissue. D Representative mIHC images showing CCR1 is expressed on macrophages in human melanoma tissues. CCR1 is indicated in red, CD68 in yellow, and DAPI in blue. Scale bars, 50 μm

Article Snippet: RAW264.7 cells were seeded in the upper chamber, and the indicated medium, supplemented with or without mCCL3 (200 nM, MCE, Cat# P7255) and the CCR1 inhibitor BX471, was carefully added to the lower chamber.

Techniques: Expressing, Gene Expression

Journal: Cell Communication and Signaling : CCS

Article Title: Blocking CCR1 + macrophages overcomes resistance to immune checkpoint inhibitors in melanoma

doi: 10.1186/s12964-025-02444-0

Figure Lengend Snippet: CCR1+ macrophages are associated with resistance to immune checkpoint blockade therapy in melanoma. A CCR1 mRNA expression in melanoma tissues compared between NR and R groups ( GSE78220 ). B CCR1 + Macrophage score in melanoma samples compared between NR and R groups ( GSE78220 ). C CCR1 mRNA expression in macrophages compared between NR and R groups ( GSE120575 ). D Number of CCR1 + cells per high-power field in melanoma tissue from Drum Tower Hospital Cohort, compared between good responders (n=5) and poor responders ( n =5). E Correlation between the CCR1 + Macrophage Score and the expression of exhausted T cell-related genes (PDCD1, HAVCR2 (TIM-3), CTLA4, and LAG3) in tumor tissues from TCGA-SKCM. Data with error bars are shown as mean ± SEM. ns, not significant; *, P <0.05; **, P <0.01; ***, P <0.001; ****, P <0.0001 as determined by unpaired Student t test ( A - D )

Article Snippet: RAW264.7 cells were seeded in the upper chamber, and the indicated medium, supplemented with or without mCCL3 (200 nM, MCE, Cat# P7255) and the CCR1 inhibitor BX471, was carefully added to the lower chamber.

Techniques: Expressing

Journal: Cell Communication and Signaling : CCS

Article Title: Blocking CCR1 + macrophages overcomes resistance to immune checkpoint inhibitors in melanoma

doi: 10.1186/s12964-025-02444-0

Figure Lengend Snippet: Synergistic therapeutic effect of BX471 (CCR1 inhibitor) combined with PD-1 blockade. A Schematic representation of syngeneic models established with B16F10 melanoma cells for combination therapy. Following tumor resection and single-cell dissociation, immune cells of interest were analyzed by flow cytometry. B Tumor growth curves of B16F10 cells treated with BX471 and PD-1 blockade (ICB) either in combination or as monotherapies. C - H ) Flow cytometry analysis of immune cell populations in tumors treated with either BX471, PD-1 blockade (ICB), a combination of both, or placebo. Cell populations analyzed include: CCR1 + macrophages ( C ); M2 macrophages ( D ); M1 macrophages ( E ); TIM3 + CD8 + T cells ( F ); PD-1 + CD8 + T cells ( G ) and CD69 + CD8 + T cells ( H ). Data with error bars are shown as mean ± SEM. ns, not significant; *, P <0.05; **, P <0.01; ***, P <0.001; ****, P <0.0001 as determined by unpaired Student t test (B-H)

Article Snippet: RAW264.7 cells were seeded in the upper chamber, and the indicated medium, supplemented with or without mCCL3 (200 nM, MCE, Cat# P7255) and the CCR1 inhibitor BX471, was carefully added to the lower chamber.

Techniques: Flow Cytometry

Journal: Cell Communication and Signaling : CCS

Article Title: Blocking CCR1 + macrophages overcomes resistance to immune checkpoint inhibitors in melanoma

doi: 10.1186/s12964-025-02444-0

Figure Lengend Snippet: Spatial niches of CCR1 + Macrophages and CD8 + Tex Cell in the melanoma tumor microenviroment (TME) and their association with ICB therapy response. A In the melanoma spatial map, the spot-level scores representing the abundance of CD8 + Tex cells and CCR1 + macrophage show a concordant pattern suggesting that the two cell types are co-localised. B Representative mIHC images demonstrating the spatial niches of CCR1 + macrophages and TIM3 + CD8 + T cells in human melanoma tissues. CCR1 is indicated in cyan, CD68 in green, CD8 in yellow, TIM3 in red and DAPI in blue. Scale bars, 20 μm. C Representative mIHC images used to calculate the nearest distance between CCR1 + macrophages and TIM3 + CD8 + T cells in human melanoma tissues. CCR1 is indicated in cyan, CD68 in green, CD8 in yellow, TIM3 in red and DAPI in blue. Scale bars, 100 μm. D Representative mIHC images demonstrating the spatial niches of CCR1 + macrophages and TIM3 + CD8 + T cells in mouse melanoma tissues from different treatment groups. CCR1 is indicated in green, F4/80 in white, CD8 in yellow, TIM3 in red and DAPI in blue. Scale bars, 100 μm. Data with error bars are shown as mean ± SEM. ns, not significant; *, P <0.05; **, P <0.01; ***, P <0.001; ****, P <0.0001 as determined by unpaired Student t test

Article Snippet: RAW264.7 cells were seeded in the upper chamber, and the indicated medium, supplemented with or without mCCL3 (200 nM, MCE, Cat# P7255) and the CCR1 inhibitor BX471, was carefully added to the lower chamber.

Techniques:

Journal: Cell Communication and Signaling : CCS

Article Title: Blocking CCR1 + macrophages overcomes resistance to immune checkpoint inhibitors in melanoma

doi: 10.1186/s12964-025-02444-0

Figure Lengend Snippet: CCL3-CCR1 axis promotes the formation of spatial niches for CCR1 + macrophages and CD8 + Tex cells. A Differential signaling ligand-receptor pairs between immune cells in responder (R) and non-responder (NR) groups. Dot sizes represent P -values (only P <0.05 are shown). B Violin plot showing the expression levels of CCR1, CCR5, CCL3, CCL3L1, CCL4, and CCL5 across different immune cell types. C Dynamic changes in the expression of CCL3, HAVCR2, PDCD1, and TCF7 during CD8 + T cell exhaustion. D Dynamic changes in the expression of CCL3, CCL4, CCL5, and CCR5 during CD8 + T cell exhaustion. E Correlation between the Exhausted T Cell Score and CCL3 expression in tumor tissues from TCGA-SKCM. F CCL3 concentration in conditioned medium of exhaustion-graded CD8⁺ T cells. G Western blot analysis of CCR1 expression in sg-CCR1 Raw264.7 cells. H Transwell assay evaluating CCL3-induced chemotactic migration of sg-CCR1 RAW264.7 cells. I Transwell to investigate chemotactic migration of RAW264.7 cells to CCL3 and the counteracting effect of CCR1 antagonist BX471. Data with error bars are shown as mean ± SEM. ns, not significant; *, P <0.05; **, P <0.01; ***, P <0.001; ****, P <0.0001 as determined by unpaired Student t test

Article Snippet: RAW264.7 cells were seeded in the upper chamber, and the indicated medium, supplemented with or without mCCL3 (200 nM, MCE, Cat# P7255) and the CCR1 inhibitor BX471, was carefully added to the lower chamber.

Techniques: Expressing, Concentration Assay, Western Blot, Transwell Assay, Migration