Journal: Discover Oncology

Article Title: CCR5 expression and conformational stability as potential cooperative modulators of immune phenotypes and therapy response in breast cancer

doi: 10.1007/s12672-025-04189-1

Figure Lengend Snippet: Comprehensive characterization of CCR5 expression, predictive significance, functional pathways, and immune contexture in breast cancer. A Expression distribution of CCR5 across normal human tissues. B Multivariate logistic regression analysis showing that high CCR5 expression is independently associated with reduced likelihood of achieving pathological complete response (pCR) after neoadjuvant chemotherapy (NAC). C Sensitivity analysis using a data-augmented cohort confirms the negative association between CCR5 and pCR, with a narrower confidence interval. D Gene Ontology (GO) gene set enrichment analysis (GSEA) reveals that CCR5-high tumors are enriched in immune-related processes including T cell activation and leukocyte adhesion, as well as cellular transport and metabolic pathways. E Hallmark GSEA indicates concurrent activation of pro-inflammatory pathways (e.g., IFN-γ response, TNFα–NF-κB, IL6–JAK–STAT3) and tumor-promoting programs such as epithelial-mesenchymal transition (EMT) and mTORC1 signaling. F KEGG pathway enrichment highlights both immunostimulatory (e.g., NK cell cytotoxicity) and protumorigenic (e.g., PI3K–Akt signaling) pathways in CCR5-high tumors. G Immune cell infiltration analysis based on multiple algorithms shows that CCR5-high tumors are associated with elevated CD8⁺ T cells, M1 macrophages, and activated dendritic cells, but also with immunosuppressive Tregs, M2 macrophages, and high stromal/immune scores, suggesting a biphasic immune microenvironment

Article Snippet: For CCR5 protein detection, we performed IHC using a rabbit anti-CCR5 polyclonal antibody (Proteintech#17476-1-AP, 1:900 dilution).

Techniques: Expressing, Functional Assay, Activation Assay, Protein-Protein interactions

Journal: Discover Oncology

Article Title: CCR5 expression and conformational stability as potential cooperative modulators of immune phenotypes and therapy response in breast cancer

doi: 10.1007/s12672-025-04189-1

Figure Lengend Snippet: Correlation between CCR5 expression and gene signatures of tumor-associated macrophage (TAM) polarization in breast cancer. A Correlation heatmap showing weak-to-moderate associations between CCR5 and M1 macrophage markers (e.g., NOS2, ARG2, PTGS2). B CCR5 expression demonstrates strong positive correlations with canonical M2 macrophage markers, including MRC1 ( r = 0.65), CD163 ( r = 0.699), and MS4A4A ( r = 0.749). C CCR5 is also highly correlated with TAM-associated immunosuppressive genes such as CD86, CCL22, and IL10, indicating a preferential link to immunoregulatory macrophage phenotypes

Article Snippet: For CCR5 protein detection, we performed IHC using a rabbit anti-CCR5 polyclonal antibody (Proteintech#17476-1-AP, 1:900 dilution).

Techniques: Expressing

Journal: Discover Oncology

Article Title: CCR5 expression and conformational stability as potential cooperative modulators of immune phenotypes and therapy response in breast cancer

doi: 10.1007/s12672-025-04189-1

Figure Lengend Snippet: Structural confidence and stability changes of CCR5 protein with V131I mutation ( A ) AlphaFold-predicted 3D structure of CCR5 highlighting residue 131. Confidence levels are color-coded based on pLDDT scores: very low (< 50), low (50–70), high (70–90), and very high (> 90). B – C Root Mean Square Fluctuation (RMSF) and Root Mean Square Deviation (RMSD) plots comparing wild-type (WT) and V131I mutant (MT) CCR5 during 100 ns molecular dynamics simulations. D Radius of gyration (Rg) trajectory indicating global compactness differences between WT and MT. E Solvent-accessible surface area (SASA) profile across simulation time. F Predicted change in protein stability (ΔΔG) upon V131I substitution at position 131, visualized as a heatmap. Red shading denotes destabilizing mutations

Article Snippet: For CCR5 protein detection, we performed IHC using a rabbit anti-CCR5 polyclonal antibody (Proteintech#17476-1-AP, 1:900 dilution).

Techniques: Mutagenesis, Residue, Solvent

Journal: International Journal of Molecular Sciences

Article Title: CCL4 Regulates Eosinophil Activation in Eosinophilic Airway Inflammation

doi: 10.3390/ijms232416149

Figure Lengend Snippet: CCL4-mediated eosinophil activation. ( A ) Purified peripheral blood eosinophils coincubated with BEAS-2B cells were treated overnight with human recombinant CCL4. CD69 expression in eosinophils was evaluated [(i) percentage of CD69+ Siglec-8+ double-positive cells and (ii) CD69 median fluorescence intensity (MFI) ratio to isotype control]. ( B ) CCL4 mRNA (i) and protein (ii) expression in eosinophils coincubated with BEAS-2B cells. ( C ) CCR5 mRNA (i) and protein (ii) expression in eosinophils coincubated with BEAS-2B cells. ( D ) Eosinophils pretreated with maraviroc (MVC)—a CCR5 antagonist—were stimulated with CCL4. The values represent the mean ± SEM of four experiments. * p < 0.05, ** p < 0.01 (between the two groups).

Article Snippet: A rabbit polyclonal antibody to CCR5 (GeneTex, Irvine, CA, USA) was used as a primary antibody, and β-actin was used as a loading control.

Techniques: Activation Assay, Purification, Recombinant, Expressing, Fluorescence, Control

Journal: International Journal of Molecular Sciences

Article Title: CCL4 Regulates Eosinophil Activation in Eosinophilic Airway Inflammation

doi: 10.3390/ijms232416149

Figure Lengend Snippet: CCL4-mediated eosinophil activation signaling. ( A ) Phosphorylation levels of multiple kinases in eosinophils. Purified peripheral blood eosinophils were treated with CCL4 (10 μg/mL) for 5 min using a Human Phospho-Kinase Antibody Array Kit (R&D). Phosphorylation levels of platelet-derived growth factor receptor (PDGFR)β, Src kinase family (Lck, Src, and Yes), extracellular signal-regulated kinase (ERK), and control are presented as dots (left panel) or a relative value to vehicle (right panel). ( B ) Colocalization of CCR5, PDGFRβ, and Src in eosinophils. CCR5 (red), PDGFRβ (green or pink), Src (light blue), and the nucleus (blue) are shown. Images were obtained using an FV3000 confocal microscope (600 × objectives). The scale bars in the bottom-right corner indicate 10 μm. The results are representative of at least three experiments. ( C ) Purified peripheral blood eosinophils were pretreated with imatinib—a PDGFRβ inhibitor (I, 5 μM) —and/or dasatinib—a Src inhibitor (D, 10 nM) —for 30 min, followed by overnight incubation with CCL4 (10 μg/mL). CD69 expression in eosinophils was determined [(i) percentage of CD69+ Siglec-8+ double-positive cells and (ii) CD69 median fluorescence intensity (MFI) ratio to isotype control]. The values represent the mean ± SEM of four experiments. # p < 0.05 (vs. nontreatment control); * p < 0.05 (between the two groups).

Article Snippet: A rabbit polyclonal antibody to CCR5 (GeneTex, Irvine, CA, USA) was used as a primary antibody, and β-actin was used as a loading control.

Techniques: Activation Assay, Phospho-proteomics, Purification, Ab Array, Derivative Assay, Control, Microscopy, Incubation, Expressing, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: CCL4 Regulates Eosinophil Activation in Eosinophilic Airway Inflammation

doi: 10.3390/ijms232416149

Figure Lengend Snippet: Mechanism of CCL4-mediated transactivation in the eosinophil signaling pathway. CCR5—a transmembrane protein coupled to G proteins that is stimulated with CCL4—induces PDGFRβ transactivation via tyrosine phosphorylation of Src, which leads to biological activities in eosinophils via signal transduction, such as kinase (e.g., ERK) phosphorylation.

Article Snippet: A rabbit polyclonal antibody to CCR5 (GeneTex, Irvine, CA, USA) was used as a primary antibody, and β-actin was used as a loading control.

Techniques: Phospho-proteomics, Transduction