Journal: Cell Death & Disease

Article Title: Deciphering the tumor immune microenvironment of imatinib-resistance in advanced gastrointestinal stromal tumors at single-cell resolution

doi: 10.1038/s41419-024-06571-3

Figure Lengend Snippet: a Cell communication analysis on TIGIT-NECTIN2 and BTLA-TNFRSF14 pair between different cell types in imatinib resistant and sensitive patients respectively. Expression of NECTIN2 ( b ), BTLA ( c ) and TNFRSF14 ( d ) in each cell type in imatinib resistant and sensitive patients respectively. e IHC analysis of NECTIN2, BTLA and TNFRSF14 between imatinib resistant (upper) and sensitive (bottom) patients. f Schematic diagram of the unique tumor-immune microenvironment of imatinib-resistance in advanced GIST.

Article Snippet: Finally, two experienced pathologists independently evaluated staining results for Foxp3 (Servicebio, GB11093), BTLA (Boster, A03149), TNFRSF14 (Affinity, AB2838686) and NECTIN2 (Boster, A08081-2).

Techniques: Expressing

Journal: Journal of Neuroinflammation

Article Title: VSIG2 as a novel immunosuppressive ligand interacts with Nectin-2 to regulate T cell responses

doi: 10.1186/s12974-025-03645-7

Figure Lengend Snippet: Nectin-2 is a functional receptor for VSIG2 on T cells. a SDS-PAGE and immunoblot analysis of T cell lysates following pull-down with hVSIG2-His protein (n =3 independent experiments). b SDS-PAGE and immunoblot analysis of CD11b + monocyte lysates following pull-down with hNectin-2-Fc protein (n =3 independent experiments). c Molecular docking model of the VSIG2-Nectin-2 interaction interface. d Key amino acid residues involved in hydrogen bond and ionic interaction formation between VSIG2 and Nectin-2, as predicted by molecular docking. e Flow cytometry analysis of Nectin-2 expression on the surface of T cells in both resting and activated states, and ( f ) statistical analysis of the results (n =3 independent experiments). g SDS-PAGE and immunoblot analysis of hVSIG2-His protein pull-down with hNectin-2-Fc protein (n =3 independent experiments). h SDS-PAGE and immunoblot analysis of hNectin-2-Fc protein pull-down with hVSIG2-His protein (n =3 independent experiments). i Isothermal titration calorimetry (ITC) curve and thermodynamic parameters for the hVSIG2-hNectin-2 binding interaction at 25°C. Data are presented as mean ± SD. Statistical significance was assessed by two-way ANOVA with Tukey’s test. Source data are provided as a Source Data file

Article Snippet: VSIG2 (Human, HEK293, His, purchased from MCE, HY- P70503 ) was dissolved in PBS buffer (pH 7.4) to a final concentration of 20 μM, and Nectin-2 (Human, HEK293, Fc, purchased from MCE, HY- P78372 ) was dissolved in the same buffer to a concentration of 200 μM.

Techniques: Functional Assay, SDS Page, Western Blot, Flow Cytometry, Expressing, Isothermal Titration Calorimetry, Binding Assay

Journal: Journal of Neuroinflammation

Article Title: VSIG2 as a novel immunosuppressive ligand interacts with Nectin-2 to regulate T cell responses

doi: 10.1186/s12974-025-03645-7

Figure Lengend Snippet: Anti-Nectin-2 Ab blocks the inhibitory effect of hVSIG2-Ig on T cells. CD3⁺ T cells were isolated by magnetic bead sorting and pre-incubated with 5 µg/ml anti-Nectin-2 antibody. Cells were then plated in 96-well plates pre-coated with anti-CD3/CD28 (1 µg/ml/0.5 µg/ml) and 6400 ng/ml hVSIG2-Ig, followed by culture for 24–72 h. a Schematic illustration of Nectin-2 receptor blockade on T cells using anti-Nectin-2 antibody. b Representative flow cytometry plots of CD69, CD25, and CD40L expression in CD4⁺ and CD8⁺ T cells, with ( c ) quantitative analysis of T cell activation ( n = 3 independent experiments). d Representative flow cytometry plots of Ki67 expression in CD4⁺ and CD8⁺ T cells, with e quantitative analysis of T cell proliferation ( n = 3 independent experiments). f Cytokine microsphere array (CBA) analysis of TNF-α, IFN-γ, IL-17 A, IL-2, and IL-6 in culture supernatants ( n = 3 independent experiments). g Immunoblot analysis of p-STAT1, GBP2, and IRF1 protein expression in T cells, with ( h ) densitometric quantification ( n = 3 independent experiments). Data are presented as mean ± SD. Statistical significance was assessed by one-way ANOVA with Tukey’s test. Source data are provided as a Source Data file

Article Snippet: VSIG2 (Human, HEK293, His, purchased from MCE, HY- P70503 ) was dissolved in PBS buffer (pH 7.4) to a final concentration of 20 μM, and Nectin-2 (Human, HEK293, Fc, purchased from MCE, HY- P78372 ) was dissolved in the same buffer to a concentration of 200 μM.

Techniques: Isolation, Incubation, Flow Cytometry, Expressing, Activation Assay, Western Blot

Journal: Journal of Neuroinflammation

Article Title: VSIG2 as a novel immunosuppressive ligand interacts with Nectin-2 to regulate T cell responses

doi: 10.1186/s12974-025-03645-7

Figure Lengend Snippet: Knockdown of Nectin-2 blocks the inhibitory effect of hVSIG2-Ig on Jurkat cells. a Schematic illustration of hVSIG2-Ig blockade in Jurkat cells with lentiviral-mediated Nectin-2 knockdown ( n = 3 independent experiments). b Flow cytometry analysis of Nectin-2 surface expression in resting and activated Jurkat cells after lentiviral transfection ( n = 3 independent experiments). c Western blot validation of Nectin-2 knockdown efficiency in Jurkat cells ( n = 3 independent experiments). d Representative flow cytometry plots of CD69, CD25, and IL-2 expression in Jurkat cells, with quantitative analysis of T cell activation markers ( n = 3 independent experiments). e Representative flow cytometry plots of Ki67 expression in Jurkat cells, with statistical analysis of proliferation rates ( n = 3 independent experiments). f Western blot analysis of p-STAT1, GBP2, and IRF1 protein levels in Jurkat cells, with g densitometric quantification of band intensities ( n = 3 independent experiments). Data are presented as mean ± SD. Statistical significance was assessed by two-way ANOVA with Tukey’s test

Article Snippet: VSIG2 (Human, HEK293, His, purchased from MCE, HY- P70503 ) was dissolved in PBS buffer (pH 7.4) to a final concentration of 20 μM, and Nectin-2 (Human, HEK293, Fc, purchased from MCE, HY- P78372 ) was dissolved in the same buffer to a concentration of 200 μM.

Techniques: Knockdown, Flow Cytometry, Expressing, Transfection, Western Blot, Biomarker Discovery, Activation Assay

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: 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