Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Immunophenotyping panel for multiplexed tissue imaging of cancer.

Article Snippet: CD112 , REA1195 , 50 , 130-122-770 , PE , Miltenyi Biotec.

Techniques: Imaging

Journal: Frontiers in Immunology

Article Title: Unveiling spatial complexity in solid tumor immune microenvironments through multiplexed imaging

doi: 10.3389/fimmu.2024.1383932

Figure Lengend Snippet: Cellular neighborhood analysis of PD1 high/low T cells in the tumor margin and core. (A–D) Topology of PD1 high (left) and PD1 low (right) T cells and their cellular neighborhood within a 5 µm range. (A, B) represent tumor margin and (C, D) show tumor core areas. Cell types showing different distribution patterns around PD1 high and PD1 low T cells (mDCs, M1-like M, M2-like M, MDSCs, Fibroblasts, vessels, tumor cells) are highlighted by arrowheads. (E, F) Quantification of cells in a 5 µm range around of PD1 high/low T cells for tumor margin and tumor core, (E) represents immune cells and (F) stroma/tumor cells. (G) Violin plots for expression levels of eight immune-modulating markers (CD112, CD155, CD276, CD39, CD73, IDO, PD-L1, and VISTA) for the most important immune and tumor cells around PD1 high/low T cells in the tumor core area. Violin plots for tumor margin are shown in Supplementary Figure S5E . Depicted markers and annotated cell types as indicated by the color code. ROI sizes: Tumor margin (ROI15) and tumor core (ROI16): 975 x 769 µm.

Article Snippet: CD112 , REA1195 , 50 , 130-122-770 , PE , Miltenyi Biotec.

Techniques: Expressing

Journal: Nature Communications

Article Title: Gut-primed neutrophils activate Kupffer cells to promote hepatic injury in mouse sepsis

doi: 10.1038/s41467-025-65572-8

Figure Lengend Snippet: WT mice were subjected to CLP and the small intestines were harvested after 20 h. A The number (no.) of neutrophils in the gut epithelium. Experiments were performed 2 times, and all data were used for analysis. Data are expressed as mean ± SEM (n = 4 samples/group) and compared by paired two-tailed Student’s t test. P value: 1.11E-2. *p < 0.05 vs. Sham. B Representative microscopic images of the gut epithelium (original magnification, ×630; scale bar: 10 μm). Experiments were performed 3 times, and representative images are shown. Bone marrow - derived neutrophils (PMNs) were treated with or without LPS in the presence and absence of intraepithelial lymphocytes (IELs) at 1:1 ratio for 12 h to evaluate NETs. C Representative microscopic images from 3 independent experiments are shown (original magnification, ×200; scale bar: 100 μm). Ly6G (purple) serves as a neutrophil marker. D Representative dot plots following the gating strategy of NETs in Fig. and E frequency of NETs + (Cit-H3 + MPO + ) neutrophils assessed by flow cytometry are shown. Experiments were performed 3 times, and all data were used for analysis. Data are expressed as mean ± SEM (n = 6 samples/group) and compared by one-way ANOVA and SNK test. P values based on the order of appearance: 3.02E-7, 1.42E-6, 4.15E-5. *p < 0.05 vs. PBS, # p < 0.05 vs. LPS, † p < 0 . 05 vs. PBS+IELs. F The number of CD112 + neutrophils in the gut epithelium of sham and CLP mice. Experiments were performed 2 times, and all data were used for analysis. Data are expressed as mean ± SEM (n = 4 samples/group) and compared by paired two-tailed Student’s t test. P value: 1.11E-2. *p < 0.05 vs. Sham. Neutrophils were incubated with IELs and LPS in the presence of a vehicle (PBS, Veh.), isotype control (Iso.), or 10 μg/mL anti-CD112 Ab for 12 h to evaluate NETs by flow cytometry. G Representative dot plots following the gating strategy of NETs in Fig. and H frequency of NETs + neutrophils are shown. Experiments were performed 2 times, and all data were used for analysis. Data are expressed as mean ± SEM (n = 5 samples/group) and compared by one-way ANOVA and SNK test. P values based on the order of appearance: 6.21E-3, 2.52E-2. *p < 0.05 vs. Veh., # p < 0.05 vs. Iso. CLP cecal ligation and puncture.

Article Snippet: Neutrophils incubated with IELs and LPS were also treated with a vehicle (PBS), isotype control, or 10 μg/mL anti-CD112 Ab (Cat. No.: MAB3869; R&D Systems, Minneapolis, MN).

Techniques: Two Tailed Test, Derivative Assay, Marker, Flow Cytometry, Incubation, Control, Ligation

Journal: Nature Communications

Article Title: Gut-primed neutrophils activate Kupffer cells to promote hepatic injury in mouse sepsis

doi: 10.1038/s41467-025-65572-8

Figure Lengend Snippet: During sepsis, neutrophils interact with IELs via CD112 in the gut, resulting in increased NETosis via PAD4-mediated histone citrullination (Cit). NET-forming neutrophils migrate from the gut into the liver and activate PAR-1 on Kupffer cells by NET-contained proteases, such as NE. NET-activated Kupffer cells polarize to an iNOS-expressing M1 phenotype and produce proinflammatory mediators, such as IL-6 and TNF. This gut-liver crosstalk mediated by immune cells leads to sepsis-induced liver injury. PAD4, protein-arginine deiminase type-4; NET, neutrophil extracellular trap; PAR-1, protease-activated receptor-1; NE, neutrophil elastase; iNOS, inducible nitric oxide synthase. Created in BioRender. Murao, A. (2025) https://BioRender.com/fhkfis3 .

Article Snippet: Neutrophils incubated with IELs and LPS were also treated with a vehicle (PBS), isotype control, or 10 μg/mL anti-CD112 Ab (Cat. No.: MAB3869; R&D Systems, Minneapolis, MN).

Techniques: Expressing

Journal: Cancers

Article Title: Targeting High-Risk Neuroblastoma Patient-Derived Xenografts with Oncolytic Virotherapy

doi: 10.3390/cancers14030762

Figure Lengend Snippet: Neuroblastoma PDXs express viral entry receptors. ( A ) Flow cytometry was used to detect the cell surface expression of the four main HSV receptors: CD111, CD112, syndecan, and HVEM in COA3, COA6, and COA129 human neuroblastoma PDX cells. All PDXs expressed all four receptors. ( B ) Representative histograms of CD111, CD112, syndecan, and HVEM cell surface expression and negative controls of COA6 are shown. Data reported as mean ± SEM and represent at least three biologic replicates.

Article Snippet: After 15 min, 10 μL of phycoerythrin (PE) conjugated anti-human CD111 antibody (Miltenyi Biotec), PE anti-human CD112 antibody (Miltenyi Biotec), allophycocyanin (APC) conjugated anti-human syndecan (Miltenyi Biotec), or APC anti-human HVEM (Miltenyi Biotec), were added for 20 min on ice in the dark.

Techniques: Flow Cytometry, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Functional tumor-reactive CD8 + T cells in pancreatic cancer

doi: 10.1186/s13046-025-03517-1

Figure Lengend Snippet: The spatial transcriptome reveals the expression patterns of NECTIN2. ( A ) H&E-stained images of sample PDACP_3 and PDACP_8. ( B ) Expression data per spatial feature was analyzed using ESTIMATE to generate a tumor purity score, stromal score, and immune score. ( C ) Spatial plots of MUC1 (Tumor marker) and NECTIN2. ( D ) Per-feature enrichment scores for CD8 T cell signature as determined by ssGSEA and Spatial plots of TIGIT. ( E ) The box plot showed the expression levels of NECTIN2 in tumor and non-tumor areas in the spatial transcriptome (Welch’s t-test, ****: P < 0.0001). ( F ) The box plot showed the expression levels of NECTIN2 in tumor and non-tumor tissues in TCGA-PDAC cohort (Mann–Whitney U test, ****: P < 0.0001). TCGA: The Cancer Genome Atlas. ( G ) Tumor sections from patients with pancreatic cancer stained with anti-PanCK (yellow), anti-NECTIN2 (red), anti-CD8(orange) and DAPI

Article Snippet: They were deparaffinized, hydrated, and manually stained with antibodies against CD8 (GB12068, Servicebio), NECTIN2 (NBP1-91211, NOVUS), TNFSF10 (MAB375, NOVUS), and PANCK ( GB122053 , Servicebio).

Techniques: Expressing, Staining, Marker, MANN-WHITNEY