murine cd274 gene  (OriGene)

Journal: Journal of Nuclear Medicine

Article Title: PD-L1 Immuno-PET Reveals Systemic Effects of Localized Oncolytic Virotherapy in a Mouse Model of Head and Neck Cancer

doi: 10.2967/jnumed.125.270922

Figure Lengend Snippet: (A) Bar charts showing PD-L1 expression via flow cytometry. (B) Confocal microscopy images of MOC1, MOC2, and MOC2( PD-L1 ) cells merged from 3 channels (Supplemental Fig. 1A). Scale = 20 µm. MFI = median fluorescence intensity.

Article Snippet: The MOC2( PD-L1 ) cell line was generated in our laboratory to overexpress PD-L1 by transducing MOC2 cells with a lentiviral vector carrying the murine CD274 gene tagged with a green fluorescent protein (GFP) reporter (MR 203953L2; OriGene Technologies).

Techniques: Expressing, Flow Cytometry, Confocal Microscopy, Fluorescence

Journal: Journal of Nuclear Medicine

Article Title: PD-L1 Immuno-PET Reveals Systemic Effects of Localized Oncolytic Virotherapy in a Mouse Model of Head and Neck Cancer

doi: 10.2967/jnumed.125.270922

Figure Lengend Snippet: (A) Saturation binding assay of 89 Zr-DFO-PD-L1 mAb on MOC2( PD-L1 ) cells. Curves for total bound (TB), specifically bound (SB), and nonspecifically bound (NSB) fractions fitted using given equation. (B) Specificity of binding assay of 89 Zr-DFO-PD-L1 mAb (2 nM; 1 h) on cancer cell lines pretreated with IFN-γ (20 ng/mL; 24 h) or PBS. Blocking with 100-fold molar excess of unlabeled PD-L1 mAb . Data were normalized to nontreated MOC2( PD-L1 ) cell–associated radioactivity. Significance determined using 2-way ANOVA with Bonferroni adjustment. C bound = bound ligand concentration; C ligand = ligand concentration; B max = maximum specific binding; K d = dissociation constant; ns = not significant.

Article Snippet: The MOC2( PD-L1 ) cell line was generated in our laboratory to overexpress PD-L1 by transducing MOC2 cells with a lentiviral vector carrying the murine CD274 gene tagged with a green fluorescent protein (GFP) reporter (MR 203953L2; OriGene Technologies).

Techniques: Saturation Assay, Binding Assay, Blocking Assay, Radioactivity, Concentration Assay

Journal: Journal of Nuclear Medicine

Article Title: PD-L1 Immuno-PET Reveals Systemic Effects of Localized Oncolytic Virotherapy in a Mouse Model of Head and Neck Cancer

doi: 10.2967/jnumed.125.270922

Figure Lengend Snippet: (A) Coregistered PET/CT scans of MOC1, MOC2, and MOC2( PD-L1 ) tumor–bearing mice 48 h after 89 Zr-DFO-PD-L1 mAb injection (PET, coronal slice; CT, 3-dimensional maximum intensity projection). (B) 89 Zr-DFO-PD-L1 mAb biodistribution profile of MOC1, MOC2, and MOC2( PD-L1 ) tumor–bearing mice 48 h after injection (110 μg; adjusted specific activity = 0.018 MBq/μg). Data pooled from 3 studies. Inset graph shows correlation of PET quantification and biodistribution (BioD). Linear regression fit with 95% CI (dashed lines). (C) PD-L1 immunohistochemistry of tumor sections. BAT = brown adipose tissue; p.i. = postinjection.

Article Snippet: The MOC2( PD-L1 ) cell line was generated in our laboratory to overexpress PD-L1 by transducing MOC2 cells with a lentiviral vector carrying the murine CD274 gene tagged with a green fluorescent protein (GFP) reporter (MR 203953L2; OriGene Technologies).

Techniques: Positron Emission Tomography-Computed Tomography, Injection, Activity Assay, Immunohistochemistry

Journal: Journal of Nuclear Medicine

Article Title: PD-L1 Immuno-PET Reveals Systemic Effects of Localized Oncolytic Virotherapy in a Mouse Model of Head and Neck Cancer

doi: 10.2967/jnumed.125.270922

Figure Lengend Snippet: Experimental timeline of 89 Zr-DFO-PD-L1 mAb immuno-PET studies to monitor PD-L1 expression after single intratumoral dose of RP1 or PBS in MOC1 tumor–bearing mice. Figure created with BioRender.com. i.v. = intravenous; s.c. = subcutaneous.

Article Snippet: The MOC2( PD-L1 ) cell line was generated in our laboratory to overexpress PD-L1 by transducing MOC2 cells with a lentiviral vector carrying the murine CD274 gene tagged with a green fluorescent protein (GFP) reporter (MR 203953L2; OriGene Technologies).

Techniques: Expressing

Journal: Journal of Nuclear Medicine

Article Title: PD-L1 Immuno-PET Reveals Systemic Effects of Localized Oncolytic Virotherapy in a Mouse Model of Head and Neck Cancer

doi: 10.2967/jnumed.125.270922

Figure Lengend Snippet: (A) Coregistered PET/CT scans of RP1 and PBS-treated mice on days 3 and 7 after treatment (PET, coronal slice; CT, 3-dimensional maximum intensity projection). Spleens of RP1-treated mice are highlighted (arrowheads). (B) Bar charts of 89 Zr-DFO-PD-L1 mAb uptake on day 3 vs. day 7 after RP1 or PBS. Significance determined using 2-way ANOVA with Bonferroni adjustment on log-transformed data. (C) Radiomics heatmaps of tumor and spleen VOIs from day 3 scans. x -axis shows feature numbers from Supplemental Tables 3 and 4. Feature values normalized to z score. Decision trees shown to left of each heatmap. TD Ln = tumor-draining lymph node.

Article Snippet: The MOC2( PD-L1 ) cell line was generated in our laboratory to overexpress PD-L1 by transducing MOC2 cells with a lentiviral vector carrying the murine CD274 gene tagged with a green fluorescent protein (GFP) reporter (MR 203953L2; OriGene Technologies).

Techniques: Positron Emission Tomography-Computed Tomography, Transformation Assay

Journal: Journal of Nuclear Medicine

Article Title: PD-L1 Immuno-PET Reveals Systemic Effects of Localized Oncolytic Virotherapy in a Mouse Model of Head and Neck Cancer

doi: 10.2967/jnumed.125.270922

Figure Lengend Snippet: (A) PD-L1 immunohistochemistry of MOC1 tumor and spleen sections from immuno-PET studies on day 3 and 7 after single intratumoral RP1 or PBS dose. (B) Concentrations of GM-CSF, IFN-α, IFN-β and IFN-γ in tumors on day 3 after RP1 or PBS. Significance determined using multiple unpaired t tests on log-transformed data with Benjamini-Hochberg FDR correction (FDR < 1%).

Article Snippet: The MOC2( PD-L1 ) cell line was generated in our laboratory to overexpress PD-L1 by transducing MOC2 cells with a lentiviral vector carrying the murine CD274 gene tagged with a green fluorescent protein (GFP) reporter (MR 203953L2; OriGene Technologies).

Techniques: Immunohistochemistry, Transformation Assay

Journal: Journal of Nuclear Medicine

Article Title: PD-L1 Immuno-PET Reveals Systemic Effects of Localized Oncolytic Virotherapy in a Mouse Model of Head and Neck Cancer

doi: 10.2967/jnumed.125.270922

Figure Lengend Snippet: (A) Representative fluorescent images of MOC1 and MOC2 cells after 48 h of incubation with increasing titers of GFP-expressing RP1-15. GFP shown in gray scale. (B) MOC1 and MOC2 cell viability after 48 h with increasing RP1 titers in vitro, assessed via CellTiter-Glo assay. Significance determined using 1-way ANOVA with Dunnett test. RLU = relative light units.

Article Snippet: The MOC2( PD-L1 ) cell line was generated in our laboratory to overexpress PD-L1 by transducing MOC2 cells with a lentiviral vector carrying the murine CD274 gene tagged with a green fluorescent protein (GFP) reporter (MR 203953L2; OriGene Technologies).

Techniques: Incubation, Expressing, In Vitro, Glo Assay

Journal: International Journal of Molecular Sciences

Article Title: Tumor-Intrinsic PD-L1 Promotes Breast Cancer Proliferation Through Livin and Galectin-1-Mediated Regulation of SKP2 Expression

doi: 10.3390/ijms27062741

Figure Lengend Snippet: Proteomic analysis of nuclear extracts from PD-L1 Cont (Sh-Cont) vs. PD-L1 KD (ShPD-L1(a) and Sh-PDL1(b)) clones of MDA-MB-231 breast cancer cells. Schematic overview of the proteomics-guided workflow used to identify PD-L1-dependent regulators of the SKP2/p21/p27 axis. The heatmap displays the top nuclear differentially expressed proteins (DEPs) in Sh-Cont compared to Sh-PD-L1(a) and Sh-PD-L1(b) (|log 2 FC| ≥ 2 and adjusted p -value ≤ 0.05). DEPs were further filtered based on literature-based relevance to proliferation, the PI3K/AKT pathway, and SKP2, p21, and/or p27 expression (details are in ).

Article Snippet: PD-L1 ORF (RC213071 from OriGene, Rockville, MD, USA) was overexpressed in MDA-MB-468 cells using Lipofectamine LTX (Thermo Fisher Scientific), followed by selection using the eukaryotic antibiotic, G 418 (Thermo Fisher Scientific) at 750 μg/mL.

Techniques: Clone Assay, Expressing

Journal: International Journal of Molecular Sciences

Article Title: Tumor-Intrinsic PD-L1 Promotes Breast Cancer Proliferation Through Livin and Galectin-1-Mediated Regulation of SKP2 Expression

doi: 10.3390/ijms27062741

Figure Lengend Snippet: Knockdown of Livin and Galectin-1 downregulates SKP2 expression and inhibits colony formation. Expression levels of SKP2 ( A ), p21 ( B ), and p27 ( C ) in MDA-MB-231 cells following transient knockdown of six candidate upstream proteins measured using qIF. Data were normalized to negative control siRNA (si-Neg) and are presented as means ± SEMs from four independent experiments. ( D ) Representative IF images (×200 magnification) showing SKP2, p21, and p27 expression after Livin or Galectin-1 knockdown compared with control siRNA. ( E ) Colony-forming ability of MDA-MB-231 cells after transient knockdown of candidate genes. Data were normalized to negative control siRNA–treated cells and are displayed as means ± SEMs from three independent experiments ( n = 3). ( F ) Representative colony-forming assays following knockdown of PD-L1, Livin, Galectin-1, EIF1AX, CALM2, and TCF-3 compared to control siRNA. Statistical significance is indicated as follows: * p < 0.05, ** p < 0.01.

Article Snippet: PD-L1 ORF (RC213071 from OriGene, Rockville, MD, USA) was overexpressed in MDA-MB-468 cells using Lipofectamine LTX (Thermo Fisher Scientific), followed by selection using the eukaryotic antibiotic, G 418 (Thermo Fisher Scientific) at 750 μg/mL.

Techniques: Knockdown, Expressing, Negative Control, Control

Journal: International Journal of Molecular Sciences

Article Title: Tumor-Intrinsic PD-L1 Promotes Breast Cancer Proliferation Through Livin and Galectin-1-Mediated Regulation of SKP2 Expression

doi: 10.3390/ijms27062741

Figure Lengend Snippet: PD-L1 promotes Livin and Galectin-1 expression in TNBC cells. ( A ) Livin and Galectin-1 expression levels in PD-L1 KD MDA-MB-231 clones compared with PD-L1 Cont controls, measured using qIF. Data were normalized to Sh-Cont MFI and are presented as means ± SEMs from four independent experiments. ( B ) Representative IF images of PD-L1 KD clones and controls (×200 magnification). ( C ) Western blot analysis of Livin and Galectin-1 expression following PD-L1 knockdown in MDA-MB-231 cells. ( D ) Densitometric quantification of the Western blots from three independent experiments presented as mean ± SEM ( n = 3). Statistical significance is indicated as follows: * p < 0.05, ** p < 0.01.

Article Snippet: PD-L1 ORF (RC213071 from OriGene, Rockville, MD, USA) was overexpressed in MDA-MB-468 cells using Lipofectamine LTX (Thermo Fisher Scientific), followed by selection using the eukaryotic antibiotic, G 418 (Thermo Fisher Scientific) at 750 μg/mL.

Techniques: Expressing, Clone Assay, Western Blot, Knockdown

Journal: International Journal of Molecular Sciences

Article Title: Tumor-Intrinsic PD-L1 Promotes Breast Cancer Proliferation Through Livin and Galectin-1-Mediated Regulation of SKP2 Expression

doi: 10.3390/ijms27062741

Figure Lengend Snippet: PD-L1 overexpression in MDA-MB-468 breast cancer cells upregulates SKP2 in a Livin/Galectin-1-dependent manner. ( A ) PD-L1 expression measured by flow cytometry following stable overexpression of PD-L1 in MDA-MB-468 cells using a PD-L1 ORF construct (PD-L1-ORF), compared with empty vector-transfected cells (vector control). ( B ) Expression of Livin, Galectin-1, and SKP2 in stable PD-L1 ORF MDA-MB-468 cells, measured using qIF. ( C ) SKP2 expression upon transient knockdown of Livin, Galectin-1, or their combination (combined) in PD-L1-ORF and vector control MDA-MB-468 cells. Data were normalized to empty vector cells and are shown as means ± SEMs from three independent experiments (right) ( n = 3). Representative IF images (×200 magnification) (left). Statistical significance is indicated as follows: * p < 0.05, *** p < 0.001, NS, not significant.

Article Snippet: PD-L1 ORF (RC213071 from OriGene, Rockville, MD, USA) was overexpressed in MDA-MB-468 cells using Lipofectamine LTX (Thermo Fisher Scientific), followed by selection using the eukaryotic antibiotic, G 418 (Thermo Fisher Scientific) at 750 μg/mL.

Techniques: Over Expression, Expressing, Flow Cytometry, Construct, Plasmid Preparation, Transfection, Control, Knockdown

Journal: International Journal of Molecular Sciences

Article Title: Tumor-Intrinsic PD-L1 Promotes Breast Cancer Proliferation Through Livin and Galectin-1-Mediated Regulation of SKP2 Expression

doi: 10.3390/ijms27062741

Figure Lengend Snippet: Knockdown of SKP2, Livin, or Galectin-1 abrogates PD-L1-promoted TNBC cell proliferation. MDA-MB-231 breast cancer cell proliferation was monitored in real time using the RTCA system. Cell index values are shown over 72 h ( upper panel , line graph) and at the 72 h endpoint ( lower panels , bar graphs; n = 3). Cells were transfected with siRNA targeting PD-L1 alone or in combination with Livin ( A ) or Galectin-1 ( B ), with or without SKP2 ( C , D ), and were compared with scrambled siRNA controls. Data are presented as mean ± SEM from at least three independent experiments ( n = 3). Statistical significance is indicated as follows: * p < 0.05, ** p < 0.01; NS, not significant.

Article Snippet: PD-L1 ORF (RC213071 from OriGene, Rockville, MD, USA) was overexpressed in MDA-MB-468 cells using Lipofectamine LTX (Thermo Fisher Scientific), followed by selection using the eukaryotic antibiotic, G 418 (Thermo Fisher Scientific) at 750 μg/mL.

Techniques: Knockdown, Transfection

Journal: International Journal of Molecular Sciences

Article Title: Tumor-Intrinsic PD-L1 Promotes Breast Cancer Proliferation Through Livin and Galectin-1-Mediated Regulation of SKP2 Expression

doi: 10.3390/ijms27062741

Figure Lengend Snippet: PD-L1 modulates Livin and Galectin-1 expression in vivo. Expression of Livin and Galectin-1 in PD-L1 Cont and PD-L1 KD MDA-MB-231 xenografts. ( Left ) Representative IF images (×100 magnification). ( Right ) Bar graphs showing expression levels (mean ± SEM), measured using qIF, normalized to Sh-Cont from three different experiments ( n = 3). (*) indicates statistical significance ( p < 0.05).

Article Snippet: PD-L1 ORF (RC213071 from OriGene, Rockville, MD, USA) was overexpressed in MDA-MB-468 cells using Lipofectamine LTX (Thermo Fisher Scientific), followed by selection using the eukaryotic antibiotic, G 418 (Thermo Fisher Scientific) at 750 μg/mL.

Techniques: Expressing, In Vivo

Journal: International Journal of Molecular Sciences

Article Title: Tumor-Intrinsic PD-L1 Promotes Breast Cancer Proliferation Through Livin and Galectin-1-Mediated Regulation of SKP2 Expression

doi: 10.3390/ijms27062741

Figure Lengend Snippet: PD-L1 upregulates Livin/Galectin-1 expression, activating the PI3K/AKT pathway to promote proliferation through SKP2–p21/p27. Schematic illustration of PD-L1-mediated PI3K/AKT activation via Livin and Galectin-1, which sustains PI3K/AKT signaling through a positive feedback loop. This PI3K/AKT activation upregulates SKP-2 and downregulates p21/p27 expression to drive cell cycle progression. Arrows indicate a promoting effect.

Article Snippet: PD-L1 ORF (RC213071 from OriGene, Rockville, MD, USA) was overexpressed in MDA-MB-468 cells using Lipofectamine LTX (Thermo Fisher Scientific), followed by selection using the eukaryotic antibiotic, G 418 (Thermo Fisher Scientific) at 750 μg/mL.

Techniques: Expressing, Activation Assay

Journal: Frontiers in Immunology

Article Title: Cell-targeted PD-1 agonists are potent NK-cell inhibitors

doi: 10.3389/fimmu.2025.1640509

Figure Lengend Snippet: PPI-ImmTAAI inhibits NK92-PD-1 cells activated with K562-HLA-A*02-PPI. (A) Representative flow cytometry profile and graphs summarising CD107a expression and IFN-γ intracellular production by NK92-PD-1 cells without (left, white circles) or with anti-PD-L1 (right, black circles), activated using different ratios of K562 or K562-PD-L1 target cells. Each circle represents the mean ( n = 6; three independent experiments; two-way ANOVA; SD; * p ≤ 0.05; *** p ≤ 0.001; **** p ≤ 0.0001). (B) Graphical representation of NK92-PD-1 activation with K562-HLA-A*02 cells loaded with the PPI 15–24 peptide in the presence of PPI-ImmTAAI. (C, D) NK92-PD-1 cells were stimulated for 4 h with K562-HLA-A*02-PPI in the presence of ctr-ImmTAAI (grey) or PPI-ImmTAAI (red). (C) Representative flow cytometry profiles and graphs summarising CD107a expression and IFN-γ intracellular production in the presence of 10 nM ImmTAAI molecules. Each dot represents one sample (mean, n = 6; three independent experiments; paired t -test; *** p ≤ 0.001; **** p ≤ 0.0001). (D) Graph summarising CD107a expression and IFN-γ intracellular production in the presence of different ImmTAAI molecules, normalised to 0 nM ImmTAAI values. Each dot represents the mean of three independent experiments ( n = 6; two-way ANOVA; SD; * p ≤ 0.05; **** p ≤ 0.0001). (E, F) NK92-PD-1 cells were stimulated for 20 h with K562-HLA-A*02-PPI in the presence of ctr-ImmTAAI (grey) or PPI-ImmTAAI (red). (E) Representative flow cytometry profiles and graph (10 nM) summarising the proportion of dead cells (PI + cells) in the presence of ImmTAAI molecules. Each dot represents one sample (mean; n = 6; three independent experiments; paired t -test; *** p ≤ 0.001). (F) Graph summarising the proportion of dead K562-HLA-A*02-PPI target cells in the presence of different concentrations of ImmTAAI, normalised to 0 nM ImmTAAI values. Each dot represents the mean of three independent experiments ( n = 6; two-way ANOVA; SD; **** p ≤ 0.0001).

Article Snippet: Lentiviral vectors containing PDCD1 (PD-1) and CD274 (PD-L1) were obtained from OriGene (Rockville, Maryland, US, RC210364L1 and RC213071L1, respectively).

Techniques: Flow Cytometry, Expressing, Activation Assay