Journal: Oncology reports

Article Title: Overexpression of B7-H1 correlates with malignant cell proliferation in pancreatic cancer.

doi: 10.3892/or.2013.2955

Figure Lengend Snippet: Figure 1. Analysis of B7-H1 expression in pancreatic carcinoma and normal pancreas tissues and validation of B7-H1 expression in stably transformed pancreatic cancer cells. (A) Immunohistochemical staining for B7-H1 expression in pancreatic carcinoma tissues. a, Negative B7-H1 staining in the normal pancreas. b, Pancreatic carcinoma tissue showed high intense staining for B7-H1 (original magnification, x400). (B) Western blot analysis of B7-H1 expression in pancreatic carcinoma specimens (lanes 1, 2, 3 and 4) and a normal pancreas specimen (lane 5). (C) Western blot analysis of B7-H1 expression levels in four pancreatic tumour cell lines: SW1990, PANC-1, MIA-PaCa-2 and BxPC-3. (D) B7-H1-transfected (B7-H1) PANC-1 clones showed increased B7-H1 expression (at both the mRNA and protein levels) compared with control cells transfected with an unmodified vector (Mock). The knockdown of B7-H1 in the BxPC-3 cell line by the lentivirus shRNA of B7-H1 significantly decreased the level of B7-H1 compared with cells treated with control shRNA. The data represent the results of three independent experiments (*P<0.05).

Article Snippet: Subsequently, the sections were incubated with a primary monoclonal antibody against B7-H1 (R&D Systems; diluted 1:50 in PBS) in a moist chamber at 4 ̊C overnight.

Techniques: Expressing, Biomarker Discovery, Stable Transfection, Transformation Assay, Immunohistochemical staining, Staining, Western Blot, Transfection, Clone Assay, Control, Plasmid Preparation, Knockdown, shRNA

Journal: Oncology reports

Article Title: Overexpression of B7-H1 correlates with malignant cell proliferation in pancreatic cancer.

doi: 10.3892/or.2013.2955

Figure Lengend Snippet: Figure 2. Effect of B7-H1 on the growth of human pancreatic tumour cells. (A) Cell growth curves of the different types of cells. An equal number of cells (2x103) were plated, and the growth rates over a period of five days were analysed by MTS assay. The data represent the results of three independent experi ments (*P<0.05). (B) Cell cycle analysis showed that the percentage of cells in the G1 phase was decreased in the B7-H1-overexpressing PANC-1 cells (Mock vs. B7-H1, 60.5±2.4 vs. 49.2±2.2%) and increased in the B7-H1-depleted BxPC-3 cells (control shRNA vs. B7-H1 shRNA, 42.3±1.4 vs. 53.8±2.4%), whereas the percentage of cells in the S phase was increased in the B7-H1-overexpressing PANC-1 cells (Mock vs. B7-H1, 31.2±1.2 vs. 35.1±1.4%) and decreased in the B7-H1-depleted BXPC-3 cells (control shRNA vs. B7-H1 shRNA, 52.9±0.9 vs. 40.9±1.3%). In addition, the percentage of cells in the G2 phase was increased in the B7-H1-overexpressing cells (Mock vs. B7-H1, 8.3±1.2 vs. 15.8±0.8%). The data represent the results of three independent experiments (*P<0.05).

Article Snippet: Subsequently, the sections were incubated with a primary monoclonal antibody against B7-H1 (R&D Systems; diluted 1:50 in PBS) in a moist chamber at 4 ̊C overnight.

Techniques: MTS Assay, Cell Cycle Assay, Control, shRNA

Journal: Oncology reports

Article Title: Overexpression of B7-H1 correlates with malignant cell proliferation in pancreatic cancer.

doi: 10.3892/or.2013.2955

Figure Lengend Snippet: Figure 3. Effect of B7-H1 on cell colony formation ability. After 12 days of culture, PANC-1 cells transfected with the B7-H1 vector formed larger and a greater number of colonies than the mock-transfected cells. In contrast, the decrease in B7-H1 expression in BxPC-3 cells led to the formation of smaller and fewer colonies when compared with the control cells. The data represent the results of three independent experiments (*P<0.05).

Article Snippet: Subsequently, the sections were incubated with a primary monoclonal antibody against B7-H1 (R&D Systems; diluted 1:50 in PBS) in a moist chamber at 4 ̊C overnight.

Techniques: Transfection, Plasmid Preparation, Expressing, Control

Journal: Oncology reports

Article Title: Overexpression of B7-H1 correlates with malignant cell proliferation in pancreatic cancer.

doi: 10.3892/or.2013.2955

Figure Lengend Snippet: Figure 4. Western blot analysis of cell cycle-related proteins in human pan creatic cancer cells. B7-H1 overexpression in the PANC-1 cell line increased the cyclin D1, CDK4/6, p-Rb and p-JNK protein levels, and the knockdown of B7-H1 in the BxPC-3 cell line decreased the levels of these proteins. GAPDH was used as a loading control.

Article Snippet: Subsequently, the sections were incubated with a primary monoclonal antibody against B7-H1 (R&D Systems; diluted 1:50 in PBS) in a moist chamber at 4 ̊C overnight.

Techniques: Western Blot, Over Expression, Knockdown, Control

Journal: Oncology reports

Article Title: Overexpression of B7-H1 correlates with malignant cell proliferation in pancreatic cancer.

doi: 10.3892/or.2013.2955

Figure Lengend Snippet: Figure 5. Real-time PCR analysis showed the alterations in the expression of epithelial-mesenchymal transition (EMT)-related molecules after the over expression of B7-H1 in PANC-1 cells. The expression of E-cadherin (E-cad), Slug, Twist, Snail and Vimentin (Vim) was analysed.

Article Snippet: Subsequently, the sections were incubated with a primary monoclonal antibody against B7-H1 (R&D Systems; diluted 1:50 in PBS) in a moist chamber at 4 ̊C overnight.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Over Expression

Journal: Radiology. Imaging cancer

Article Title: PET/MRI and Bioluminescent Imaging Identify Hypoxia as a Cause of Programmed Cell Death Ligand 1 Image Heterogeneity.

doi: 10.1148/rycan.220138

Figure Lengend Snippet: Figure 1: Increased expression of luciferase and programmed cell death ligand 1 (PD-L1) in 4T1-HRE-ODD-Luc cells exposed to hypoxia. (A) Graph shows the fold increase of luciferase messenger RNA (mRNA) in cells exposed to 1% oxygen compared with cells exposed to 20% oxygen for 24 hours. (B) Graph shows the fold increase of luciferase activity in cells exposed to 1% oxygen compared with cells exposed to 20% oxygen for 24 hours. (C) Graph shows the fold increase of PD-L1 mRNA in cells exposed to 1% oxygen compared with cells exposed to 20% oxygen for 24 hours. (D) Immunoblots show PD-L1 protein expression in cells exposed to 20% or 1% oxygen for 24 hours and 48 hours. mRNA experiments were performed in triplicate. Central horizontal lines represent the geometric mean, bars represent ± SD, and dots represent individual samples. GAPDH = glyceraldehyde 3-phosphate dehydrogenase, HRE = hypoxia response element, Luc = luciferase, ODD = oxygen-dependent degradation. * P ≤ .05, *** P ≤ .0005, **** P ≤ .00005.

Article Snippet: A monoclonal primary antibody against human PD-L1 for the human cell line (1:1000; GeneTex), a goat polyclonal antibody against mouse PD-L1 (1:500, mouse B7-1H, catalog no. AF1019; R&D Systems) for the mouse cell line, and an anti–glyceraldehyde 3-phosphate dehydrogenase antibody (mouse monoclonal; Sigma) were used.

Techniques: Expressing, Luciferase, Activity Assay, Western Blot

Journal: Radiology. Imaging cancer

Article Title: PET/MRI and Bioluminescent Imaging Identify Hypoxia as a Cause of Programmed Cell Death Ligand 1 Image Heterogeneity.

doi: 10.1148/rycan.220138

Figure Lengend Snippet: Figure 2: Increased programmed cell death ligand 1 (PD-L1) expression in SUM149 human breast cancer cells exposed to hypoxia. (A) Graph shows the fold increase of PD-L1 messenger RNA (mRNA) in cells exposed to 1% oxygen compared with cells exposed to 20% oxygen for 24 hours. (B) Immunoblots show PD-L1 protein expression in cells exposed to 20% or 1% oxygen for 24 hours and 48 hours. mRNA experiments were performed in triplicate. Central horizontal lines represent the geometric mean, bars represent ± SD, and dots repre- sent individual samples. GAPDH = glyceraldehyde 3-phosphate dehydrogenase. * P ≤ .05.

Article Snippet: A monoclonal primary antibody against human PD-L1 for the human cell line (1:1000; GeneTex), a goat polyclonal antibody against mouse PD-L1 (1:500, mouse B7-1H, catalog no. AF1019; R&D Systems) for the mouse cell line, and an anti–glyceraldehyde 3-phosphate dehydrogenase antibody (mouse monoclonal; Sigma) were used.

Techniques: Expressing, Western Blot

Journal: Radiology. Imaging cancer

Article Title: PET/MRI and Bioluminescent Imaging Identify Hypoxia as a Cause of Programmed Cell Death Ligand 1 Image Heterogeneity.

doi: 10.1148/rycan.220138

Figure Lengend Snippet: Figure 3: Spatial colocalization between high bioluminescence imaging (BLI) intensity and high programmed cell death ligand 1 (PD-L1) image intensity in tumors in vivo. (A) Representative BL image of mouse with 4T1-HRE-Luc tumor obtained immediately before start of PET/MRI studies shows high BLI intensity in the hypoxic tumor region. (B) Representative PET/MR images from the same mouse obtained at 24 hours and at 48 hours after injection. HRE = hypoxia response element, Luc = luciferase, max = maximum, min = minimum, p/s/cm2/sr = photon per square centimeter per second per steradian.

Article Snippet: A monoclonal primary antibody against human PD-L1 for the human cell line (1:1000; GeneTex), a goat polyclonal antibody against mouse PD-L1 (1:500, mouse B7-1H, catalog no. AF1019; R&D Systems) for the mouse cell line, and an anti–glyceraldehyde 3-phosphate dehydrogenase antibody (mouse monoclonal; Sigma) were used.

Techniques: Imaging, In Vivo, Positron Emission Tomography-Magnetic Resonance Imaging, Injection, Luciferase

Journal: Radiology. Imaging cancer

Article Title: PET/MRI and Bioluminescent Imaging Identify Hypoxia as a Cause of Programmed Cell Death Ligand 1 Image Heterogeneity.

doi: 10.1148/rycan.220138

Figure Lengend Snippet: Figure 4: Spatial colocalization between ex vivo high bioluminescence imaging (BLI) intensity and high programmed cell death ligand 1 (PD-L1) image intensity in corresponding high-resolution PET image. (A) Bright-field image and (B) BL image obtained from the central slice of the tumor displayed in Figure 3. Corresponding colo- calized high-resolution (C) MR, (D) PET, and (E) PET/MR images obtained in vivo. (F) Representative low-magnification (×1) immunohistochemical image for PD-L1 with distinct necrotic areas (N). Magnified regions of two different areas (inset) show necrotic regions and immediately adjacent viable cells with intense membrane staining for PD-L1. max = maximum, min = minimum, p/s/cm2/sr = photon per square centimeter per second per steradian.

Article Snippet: A monoclonal primary antibody against human PD-L1 for the human cell line (1:1000; GeneTex), a goat polyclonal antibody against mouse PD-L1 (1:500, mouse B7-1H, catalog no. AF1019; R&D Systems) for the mouse cell line, and an anti–glyceraldehyde 3-phosphate dehydrogenase antibody (mouse monoclonal; Sigma) were used.

Techniques: Ex Vivo, Imaging, Positron Emission Tomography-Magnetic Resonance Imaging, In Vivo, Immunohistochemical staining, Membrane, Staining

Journal: Radiology. Imaging cancer

Article Title: PET/MRI and Bioluminescent Imaging Identify Hypoxia as a Cause of Programmed Cell Death Ligand 1 Image Heterogeneity.

doi: 10.1148/rycan.220138

Figure Lengend Snippet: Figure 5: Relating hypoxic tumor areas to PET image intensity in colocalized regions. (A) Schematic shows the workflow using ex vivo bioluminescence images to classify regions with 0%–25%, 25%–50%, 50%–75%, and 75%–100% signal intensities that were mapped onto the corresponding colocalized PET/MR images to obtain programmed cell death ligand 1 (PD-L1) signal intensity corresponding to those regions. (B) Graph shows a summary of the data obtained from analyzing the central slice of six tumors. Significantly higher PD-L1 expression was observed in regions with the highest (75%–100%) and next highest (50%–75%) bioluminescence imaging (BLI) intensity compared with the region with lowest (0%–25%) BLI intensity. Central horizontal lines represent the geometric mean, bars represent ± SD, and dots represent indi- vidual samples. # P ≤ .06, ** P ≤ .001. AU = arbitrary unit.

Article Snippet: A monoclonal primary antibody against human PD-L1 for the human cell line (1:1000; GeneTex), a goat polyclonal antibody against mouse PD-L1 (1:500, mouse B7-1H, catalog no. AF1019; R&D Systems) for the mouse cell line, and an anti–glyceraldehyde 3-phosphate dehydrogenase antibody (mouse monoclonal; Sigma) were used.

Techniques: Ex Vivo, Positron Emission Tomography-Magnetic Resonance Imaging, Expressing, Imaging

Journal: Radiology. Imaging cancer

Article Title: PET/MRI and Bioluminescent Imaging Identify Hypoxia as a Cause of Programmed Cell Death Ligand 1 Image Heterogeneity.

doi: 10.1148/rycan.220138

Figure Lengend Snippet: Figure 6: Programmed cell death ligand 1 (PD-L1) messenger RNA expression in treatment-naive triple-negative breast cancer (TNBC) with high and low hypoxia- inducible factor (HIF)–1α expression. Comparison of RNA sequencing data, expressed as log2 based on the RNA-Seq version 2 by expectation maximization (RNA SeqV2RSEM) method of analysis, of PD-L1 in TNBC patient samples with high or low HIF-1α expression from three different studies. Data represent geometric means with 95% CIs from 10 or more samples (dots) in each study. Statistical analysis was performed with the Mann-Whitney U test using Prism software (GraphPad). BIC = breast invasive carcinoma, CD274 = cluster of differentiation 274, SMC = Samsung Medical Center, TCGA = The Cancer Genome Atlas. * P ≤ .05.

Article Snippet: A monoclonal primary antibody against human PD-L1 for the human cell line (1:1000; GeneTex), a goat polyclonal antibody against mouse PD-L1 (1:500, mouse B7-1H, catalog no. AF1019; R&D Systems) for the mouse cell line, and an anti–glyceraldehyde 3-phosphate dehydrogenase antibody (mouse monoclonal; Sigma) were used.

Techniques: RNA Expression, Expressing, Comparison, RNA Sequencing, MANN-WHITNEY, Software

Journal: International Journal of Medical Sciences

Article Title: Arbutin overcomes tumor immune tolerance by inhibiting tumor programmed cell death-ligand 1 expression

doi: 10.7150/ijms.92419

Figure Lengend Snippet: Arbutin inhibited PD-L1 expression in B16F10 and LL2 cells. Arbutin reduced PD-L1 expression in B16F10 and LL2 cells. (A) B16F10 and LL2 cells (5 × 10 5 cells/well) were placed into 6-well plates and incubated at 37 °C for 24 h. Then treatment with Salmonella (5 × 10 7 cells/well) for 1.5h or CoCl 2 (200 μM) for 6 h, the expression of PD-L1 was measured by Western blotting. B16F10 (B) and LL2 (C) cells (5 × 10 5 cells/well) were placed into 6-well plates and incubated at 37 °C for 24 h. Then treatment with arbutin (0-1.56 μM) for 6 h, the expression of PD-L1 was measured by Western blotting. Arbutin reduced PD-L1 expression on the surface of cells. After treatment with arbutin (1.56 μM) for 6 h, the expression of PD-L1 on the surface of B16F10 (C) and LL2 (D) cells was measured by flow cytometry. (mean ± SD, n=5; *, p < 0.05).

Article Snippet: The membrane was then probed with the following primary antibodies: PD-L1 (Dilution: 1:1000) (GeneTex), phospho-AKT (Dilution: 1:1000) (Santa Cruz Biotechnology Inc, Santa Cruz, CA, USA), AKT (Dilution: 1:1000) (Santa Cruz Biotechnology), phospho-mTOR (Dilution: 1:1000) (Cell Signaling, Danvers, MA, USA), mTOR (Dilution: 1:1000) (Cell Signaling), caspase 3 (Dilution: 1:1000) (GeneTex), LC3 (Dilution: 1:1000) (Novus Biologicals, Littleton, CO, USA) and a monoclonal antibody against β-actin (Dilution: 1:10000) (Sigma-Aldrich).

Techniques: Expressing, Incubation, Western Blot, Flow Cytometry

Journal: International Journal of Medical Sciences

Article Title: Arbutin overcomes tumor immune tolerance by inhibiting tumor programmed cell death-ligand 1 expression

doi: 10.7150/ijms.92419

Figure Lengend Snippet: Arbutin-mediated PD-L1 protein expression. B16F10 (A) and LL2 (B) cells (5 × 10 5 cells/well) were placed into 6-well plates and incubated at 37 °C for 24 h. Then treatment with arbutin (0-1.56 μM) for 6 h, the expression of PD-L1 and phosphorylation-AKT/mTOR were measured by Western blotting. Quantification histograms are presented beneath each Western blotting plot. Data are expressed as the mean ± SD of three-time repeated determinations. Each experiment was repeated three times with similar results.

Article Snippet: The membrane was then probed with the following primary antibodies: PD-L1 (Dilution: 1:1000) (GeneTex), phospho-AKT (Dilution: 1:1000) (Santa Cruz Biotechnology Inc, Santa Cruz, CA, USA), AKT (Dilution: 1:1000) (Santa Cruz Biotechnology), phospho-mTOR (Dilution: 1:1000) (Cell Signaling, Danvers, MA, USA), mTOR (Dilution: 1:1000) (Cell Signaling), caspase 3 (Dilution: 1:1000) (GeneTex), LC3 (Dilution: 1:1000) (Novus Biologicals, Littleton, CO, USA) and a monoclonal antibody against β-actin (Dilution: 1:10000) (Sigma-Aldrich).

Techniques: Expressing, Incubation, Phospho-proteomics, Western Blot

Journal: International Journal of Medical Sciences

Article Title: Arbutin overcomes tumor immune tolerance by inhibiting tumor programmed cell death-ligand 1 expression

doi: 10.7150/ijms.92419

Figure Lengend Snippet: Arbutin reduces PD-L1 expression through the AKT/mTOR pathway. B16F10 (A) and LL2 (B) cells (5 × 10 5 cells/well) were placed into 6-well plates and incubated at 37 °C for 24 h. Transfect the cells with control or constitutively active AKT plasmid (5ug) at 37°C for 6 h, then treat with arbutin (1.56 μM) for 6 h. Analyze the expression levels of the AKT/mTOR proteins and PD-L1 in the cells using Western blotting. Quantification histograms are presented beneath each Western blotting plot. Data are expressed as the mean ± SD of three-time repeated determinations. Each experiment was repeated three times with similar results.

Article Snippet: The membrane was then probed with the following primary antibodies: PD-L1 (Dilution: 1:1000) (GeneTex), phospho-AKT (Dilution: 1:1000) (Santa Cruz Biotechnology Inc, Santa Cruz, CA, USA), AKT (Dilution: 1:1000) (Santa Cruz Biotechnology), phospho-mTOR (Dilution: 1:1000) (Cell Signaling, Danvers, MA, USA), mTOR (Dilution: 1:1000) (Cell Signaling), caspase 3 (Dilution: 1:1000) (GeneTex), LC3 (Dilution: 1:1000) (Novus Biologicals, Littleton, CO, USA) and a monoclonal antibody against β-actin (Dilution: 1:10000) (Sigma-Aldrich).

Techniques: Expressing, Incubation, Control, Plasmid Preparation, Western Blot

Journal: International Journal of Medical Sciences

Article Title: Arbutin overcomes tumor immune tolerance by inhibiting tumor programmed cell death-ligand 1 expression

doi: 10.7150/ijms.92419

Figure Lengend Snippet: Arbutin inhibits tumor growth and PD-L1 expression in vivo . B16F10 and LL2 tumor cells were subcutaneously injected into C57BL/6 mice on day 0. Tumor growth was allowed for seven days, and arbutin (50 mg/kg) was administered via intraperitoneal injection for seven consecutive days from day 8. Tumor volumes were measured every three days. On Day 15, tumors were extracted using a lysis buffer. The supernatant was collected, and PD-L1 expression levels were analyzed using Western blotting for (A) B16F10 (n=3) and (B) LL2 (n=3) tumors. Tumor volumes were compared between the control group and the arbutin-treated group for (C) B16F10 (n=10) and (D) LL2 (n=9) tumors. (mean ± SEM; *, p < 0.05).

Article Snippet: The membrane was then probed with the following primary antibodies: PD-L1 (Dilution: 1:1000) (GeneTex), phospho-AKT (Dilution: 1:1000) (Santa Cruz Biotechnology Inc, Santa Cruz, CA, USA), AKT (Dilution: 1:1000) (Santa Cruz Biotechnology), phospho-mTOR (Dilution: 1:1000) (Cell Signaling, Danvers, MA, USA), mTOR (Dilution: 1:1000) (Cell Signaling), caspase 3 (Dilution: 1:1000) (GeneTex), LC3 (Dilution: 1:1000) (Novus Biologicals, Littleton, CO, USA) and a monoclonal antibody against β-actin (Dilution: 1:10000) (Sigma-Aldrich).

Techniques: Expressing, In Vivo, Injection, Lysis, Western Blot, Control

Journal: Frontiers in Medicine

Article Title: Serum sodium levels associate with recovery of kidney function in immune checkpoint inhibitor nephrotoxicity

doi: 10.3389/fmed.2023.1020691

Figure Lengend Snippet: Clinical, laboratory, and histopathological parameters of the total cohort.

Article Snippet: After deparaffinization in xylene and rehydration in ethanol containing distilled water, formalin-fixed, paraffin-embedded kidney sections were stained using primary antibodies against PD-L1 (1:100, ab205921, Abcam, Cambridge, UK), PD-1 (1:500, ab52587, Abcam, Cambridge, UK), C1q (1:30,000, A0136, Agilent Dako, Santa Clara, USA), and C3c (1:10,000, A0062, Agilent Dako, Santa Clara, USA), labeling was performed using Novolink™ Polymer Detection System (Leica Biosystems, Wetzlar, Germany) according to the manufacturer’s protocol.

Techniques:

Journal: Frontiers in Medicine

Article Title: Serum sodium levels associate with recovery of kidney function in immune checkpoint inhibitor nephrotoxicity

doi: 10.3389/fmed.2023.1020691

Figure Lengend Snippet: Histopathological lesions or cumulative steroid dose do not affect renal recovery after AIN related to ICI nephrotoxicity. (A) Difference between worst eGFR at time of diagnosis and last documented eGFR during follow-up in the patient cohort are shown. (B) Intensity of PD-L1/PD-1 positivity within different renal compartments, cumulative steroid dose and histopathological findings in association with eGFR recovery in AIN related to ICI nephrotoxicity are shown by heatmap reflecting mean values of Spearman’s ρ. AIN, acute interstitial nephritis; eGFR, estimated glomerular filtration rate (CKD-EPI); ICI, immune checkpoint inhibitor; IF/TA, interstitial fibrosis/tubular atrophy; PD-1, programmed cell death protein 1; PD-L1, programmed cell death protein 1-ligand 1.

Article Snippet: After deparaffinization in xylene and rehydration in ethanol containing distilled water, formalin-fixed, paraffin-embedded kidney sections were stained using primary antibodies against PD-L1 (1:100, ab205921, Abcam, Cambridge, UK), PD-1 (1:500, ab52587, Abcam, Cambridge, UK), C1q (1:30,000, A0136, Agilent Dako, Santa Clara, USA), and C3c (1:10,000, A0062, Agilent Dako, Santa Clara, USA), labeling was performed using Novolink™ Polymer Detection System (Leica Biosystems, Wetzlar, Germany) according to the manufacturer’s protocol.

Techniques: Filtration