Journal: Science Advances

Article Title: Nanocollision promotes locomotion of dendritic cells for tumor therapy

doi: 10.1126/sciadv.aeb7714

Figure Lengend Snippet: ( A ) Synthesis process of an NCG. (PFH, liquid-gas phase-change core; PDA, stabilizer; FA, solid-liquid phase-change shell) capable of gas-liquid-solid triphasic conversion. ( B ) Schematic of the strategy for NCG-triggered nanocollision for enhanced locomotion of DCs. First, ultrasound induces vaporization of PFH. Then, the gasified PFH generates a critical internal pressure, triggering instantaneous rupture of the FA shell with subsequent outward ejection of fragmented particulates. Subsequently, the fragmented particulates induce nanocollisions with DCs, eliciting localized fluctuation of plasma membrane. IV, Piezo1 detects the fluctuation and mediates Ca 2+ influx through its central pore. Finally, Ca 2+ influx induces F-actin polymerization (enhanced intrinsic locomotion) and high expression of CCR7 (enhanced chemotaxis). ( C ) Locomotion-enhanced DCs potentiate antigen capture and lymph node homing, thereby activating T cells to amplify antitumor immunity.

Article Snippet: Antibodies and reagents used for Western blot are as follows: rabbit anti–IL-12 (bs-0767R), rabbit anti–IFN-γ (bs-0480R), CCR7 rabbit polyclonal antibody (pAb) (bs-1305R), and β-actin mouse monoclonal antibody (bsm-33036 M) from Bioss; cPLA2 pAb (YT1084), cPLA2 (phospho Ser 505 ) pAb (YP0868), PIEZ1 rabbit pAb (YT8073), AP-1 (phospho Tyr 170 ) pAb (YP0018), and AP-1 (Acetyl Lys271) pAb (YK0062) from Immunoway; anti-calreticulin rabbit pAb ( GB112134 ), anti-HMGB1 rabbit pAb (GB11103) from Servicebio.

Techniques: Clinical Proteomics, Membrane, Expressing, Chemotaxis Assay

Journal: Science Advances

Article Title: Nanocollision promotes locomotion of dendritic cells for tumor therapy

doi: 10.1126/sciadv.aeb7714

Figure Lengend Snippet: ( A and B ) Representative flow cytometry dot plots (A) and percentage (B) of CFSE-stained DCs in adjacent lymph nodes. ( n = 5). ( C ) Western blot analysis of CCR7 expression and oligomerization. ( D ) cPLA 2 pathway–related gene alterations, heatmap. (C versus B). ( E and F ) Western blot analysis of expression for proteins in the cPLA 2 pathway, including Piezo1, p-cPLA 2 , CCR7, and their quantitative analysis. ( G ) Representative images of Ca 2+ diffusion from the protrusions to the cell interior. ( H ) Schematic illustration of Ca 2+ influx–induced CCR7 expression in DC. ( I ) Heatmap of collagen and integrin-related genes (A versus B versus C). ( J ) Sankey bubble plot of pathway changes corresponding to genes. (C versus B). ( K ) Density plot of RNA-seq. (A versus B versus C). ( L ) Bubble plot of nanocollision-mediated alterations in DC signaling cascades. (C versus B). ( M and N ) Representative fluorescence images (M) and statistical graphs (N) of deep infiltration of DCs. (A, control; B, magnetic nanospheres; C, magnetic nanospheres with magnetic field, which can achieve collision. n = 6) ( P values: ns, not significant, * P < 0.05, ** P < 0.01.)

Article Snippet: Antibodies and reagents used for Western blot are as follows: rabbit anti–IL-12 (bs-0767R), rabbit anti–IFN-γ (bs-0480R), CCR7 rabbit polyclonal antibody (pAb) (bs-1305R), and β-actin mouse monoclonal antibody (bsm-33036 M) from Bioss; cPLA2 pAb (YT1084), cPLA2 (phospho Ser 505 ) pAb (YP0868), PIEZ1 rabbit pAb (YT8073), AP-1 (phospho Tyr 170 ) pAb (YP0018), and AP-1 (Acetyl Lys271) pAb (YK0062) from Immunoway; anti-calreticulin rabbit pAb ( GB112134 ), anti-HMGB1 rabbit pAb (GB11103) from Servicebio.

Techniques: Flow Cytometry, Staining, Western Blot, Expressing, Diffusion-based Assay, RNA Sequencing, Fluorescence, Control

Journal: Science Advances

Article Title: Nanocollision promotes locomotion of dendritic cells for tumor therapy

doi: 10.1126/sciadv.aeb7714

Figure Lengend Snippet: ( A and B ) Representative fluorescence images of intracellular Ca 2+ distribution in DCs with different treatments. ( C to E ) Locomotion trajectory (C), accumulated distance (in 5 min) (D), and locomotion speed (E) of DCs with different treatments. ( F and G ) Western blot analysis of the nanocollision-induced alterations of monomeric and oligomeric CCR7 (F) and Piezo1 expression (G). ( H and I ), Representative fluorescence images showing the antigen capture (H) and antigen presenting (I) capabilities of DCs with different treatments. ( J ) Western blot analysis of IL-12 and IFN-γ expression. ( K to N ) Transcriptomics analysis of gene expression in DCs. RNA-seq density plot [(K), A versus B versus C], heatmap of immune activation-related genes [(L) A versus B versus C], bubble plot [(M) C versus B], and sankey bubble plot [(N) C versus B]. [(A) Control; (B) R848; (C) R848 + NCG US(40°C) ]. P values: ns, not significant, * P < 0.05, ** P < 0.01, **** P < 0.0001.)

Article Snippet: Antibodies and reagents used for Western blot are as follows: rabbit anti–IL-12 (bs-0767R), rabbit anti–IFN-γ (bs-0480R), CCR7 rabbit polyclonal antibody (pAb) (bs-1305R), and β-actin mouse monoclonal antibody (bsm-33036 M) from Bioss; cPLA2 pAb (YT1084), cPLA2 (phospho Ser 505 ) pAb (YP0868), PIEZ1 rabbit pAb (YT8073), AP-1 (phospho Tyr 170 ) pAb (YP0018), and AP-1 (Acetyl Lys271) pAb (YK0062) from Immunoway; anti-calreticulin rabbit pAb ( GB112134 ), anti-HMGB1 rabbit pAb (GB11103) from Servicebio.

Techniques: Fluorescence, Western Blot, Expressing, Gene Expression, RNA Sequencing, Activation Assay, Control

Journal: International Dental Journal

Article Title: CCR7 Promotes Neutrophil Recruitment and N2 Polarization Through Regulation of IL-16 in Head and Neck Squamous Cell Carcinoma

doi: 10.1016/j.identj.2025.109311

Figure Lengend Snippet: Knockdown of CCR7 in HNSCC restrains neutrophil recruitment. A, Assessment of CCR7 mRNA expression levels in five HNSCC cell lines via qRT‒PCR. B, Representative fluorescence images of the morphology of FADU and PCI-37B cells after siRNA transfection. FAM-labelled siRNA is shown in green (scale bar: 200 μm). C, The knockdown efficiency of siCCR7 was verified via qRT‒PCR in the FADU and PCI-37B cell lines. D, The proportion of CD11b + neutrophils induced by DMSO in HL-60 cells was examined by flow cytometry. E, Assessment of the effect of CM from HNSCC cells on neutrophil migration capability via Transwell migration assays (scale bar: 200 μm). The histogram shows the statistical results of the flow cytometry counts. F, Assessment of the effect of CM from HNSCC cells on neutrophil invasion ability via Transwell invasion assays (scale bar: 200 μm). The histogram shows the statistical results of the flow cytometry counts. * P < .05, ** P < .01, *** P < .001, **** P < .0001.

Article Snippet: CCL19, CCL21 (PeproTech, USA); CCR7 monoclonal antibody (BOSTER, China); FITC anti-mouse/human CD11b antibody, PE anti-mouse CD206 (MMR) antibody (BioLegend, USA); human IL-16 antibody (Affinity Biosciences, Australia); and CD206 rabbit polyclonal antibody (ABclonal, China).

Techniques: Knockdown, Expressing, Fluorescence, Transfection, Flow Cytometry, Migration

Journal: International Dental Journal

Article Title: CCR7 Promotes Neutrophil Recruitment and N2 Polarization Through Regulation of IL-16 in Head and Neck Squamous Cell Carcinoma

doi: 10.1016/j.identj.2025.109311

Figure Lengend Snippet: CCR7 overexpression is significantly correlated with neutrophil infiltration and the N2 phenotype in patients with HNSCC. A, CCR7 expression patterns of different tumour types in TIMER 2.0. The red boxes represent HNSCC tissues and adjacent normal tissues. B, CCR7 expression in HNSCC patients and healthy controls in the TCGA cohort. C, Protein expression levels of CCR7 in HNSCC tissues and normal oral epithelial mucosal epithelial tissues from the HPA, magnification, × 4. D, Immune infiltration levels in HNSCC patients stratified by CCR7 expression levels, utilizing data from the TCGA. E, Neutrophil infiltration levels in HNSCC were compared between the CCR7 high- and low-expression groups via QUANTISEQ. F, Correlation between CCR7 mRNA expression levels and neutrophil infiltration according to the TIMER algorithm. G, Spearman correlation analysis was used to assess the relationship between CCR7 mRNA expression levels and N2 neutrophil marker (MRC1, CCL2) mRNA expression via TIMER 2.0. * P < .05, ** P < .01, *** P < .001.

Article Snippet: CCL19, CCL21 (PeproTech, USA); CCR7 monoclonal antibody (BOSTER, China); FITC anti-mouse/human CD11b antibody, PE anti-mouse CD206 (MMR) antibody (BioLegend, USA); human IL-16 antibody (Affinity Biosciences, Australia); and CD206 rabbit polyclonal antibody (ABclonal, China).

Techniques: Over Expression, Expressing, Marker

Journal: International Dental Journal

Article Title: CCR7 Promotes Neutrophil Recruitment and N2 Polarization Through Regulation of IL-16 in Head and Neck Squamous Cell Carcinoma

doi: 10.1016/j.identj.2025.109311

Figure Lengend Snippet: Knockdown of CCR7 inhibits N2 polarization of neutrophils in HNSCC. A, Morphological changes in neutrophil nuclei following coculture with various CMs were visualized via Wright‒Giemsa staining (scale bar: 100 μm). B, The nuclei were stained with DAPI (blue), while the cytoskeleton was labelled with Actin-Tracker Red-555 fluorescent phalloidin (red) (scale bar: 20 μm). C, The influence of HNSCC cells on the relative expression levels of neutrophil N2 markers was evaluated via qRT‒PCR. D, The proportion of CD206-positive neutrophils after coculture was detected by flow cytometry. E, Representative fluorescence images of neutrophils. The cell membrane CXCR2 was labelled with Alexa Fluor® 488 (green), and the nuclei were stained with DAPI (blue) (scale bar: 50 μm). F, Histogram of the results of the statistical analysis of the CXCR2 fluorescence intensity values on the membrane surface of neutrophils after coculture with the FADU or PCI-37B. G, Neutrophil CCL2 secretion levels were quantified via ELISA after co-culturing with FADU and PCI-37B. * P < .05, ** P < .01, *** P < .001.

Article Snippet: CCL19, CCL21 (PeproTech, USA); CCR7 monoclonal antibody (BOSTER, China); FITC anti-mouse/human CD11b antibody, PE anti-mouse CD206 (MMR) antibody (BioLegend, USA); human IL-16 antibody (Affinity Biosciences, Australia); and CD206 rabbit polyclonal antibody (ABclonal, China).

Techniques: Knockdown, Staining, Expressing, Flow Cytometry, Fluorescence, Membrane, Enzyme-linked Immunosorbent Assay

Journal: International Dental Journal

Article Title: CCR7 Promotes Neutrophil Recruitment and N2 Polarization Through Regulation of IL-16 in Head and Neck Squamous Cell Carcinoma

doi: 10.1016/j.identj.2025.109311

Figure Lengend Snippet: CCR7 regulates IL-16 expression in HNSCC. A, Genes with the most significant correlation with CCR7 mRNA expression patterns in HNSCC according to the UALCAN database were analysed and identified. B, Correlation analysis of CCR7 and IL-16 mRNA expression in HNSCC, based on the UALCAN database. C, Spearman correlation analysis of CCL19, CCL21 and IL-16 in HNSCC based on the TIMER2.0 database. D, Immunofluorescence was performed on paraffin sections of cancerous and paracancerous tissues from HNSCC patients; CCR7 was labelled with Alexa Fluor® 488 (green), IL-16 with Alexa Fluor® 594 (red), and nuclei with DAPI (blue) (scale bar: 200 μm). E, IL-16 mRNA expression was measured via qRT‒PCR after CCR7 was knocked down in the FADU and PCI-37B. F, IL-16 protein secretion was measured by ELISA after CCR7 knockdown in the FADU and PCI-37B cells. G, IL-16 mRNA expression was detected in FADU and PCI-37B groups following CCL19/21 addition, while the control group was pretreated with CCR7 mAb. H, IL-16 protein secretion was detected in the FADU and PCI-37B groups following CCL19/21 addition, while the control group was pretreated with a CCR7 mAb. * P < .05, ** P < .01, *** P < .001, **** P < .0001.

Article Snippet: CCL19, CCL21 (PeproTech, USA); CCR7 monoclonal antibody (BOSTER, China); FITC anti-mouse/human CD11b antibody, PE anti-mouse CD206 (MMR) antibody (BioLegend, USA); human IL-16 antibody (Affinity Biosciences, Australia); and CD206 rabbit polyclonal antibody (ABclonal, China).

Techniques: Expressing, Immunofluorescence, Enzyme-linked Immunosorbent Assay, Knockdown, Control