Journal: Frontiers in Immunology

Article Title: Low-Dose Gemcitabine Treatment Enhances Immunogenicity and Natural Killer Cell-Driven Tumor Immunity in Lung Cancer

doi: 10.3389/fimmu.2020.00331

Figure Lengend Snippet: List of primers used in qRT-PCR.

Article Snippet: Anti-human ULBP1, ULBP2/5/6, and ULBP3 antibody were purchased from R&D Systems.

Techniques:

Journal: Frontiers in Immunology

Article Title: Low-Dose Gemcitabine Treatment Enhances Immunogenicity and Natural Killer Cell-Driven Tumor Immunity in Lung Cancer

doi: 10.3389/fimmu.2020.00331

Figure Lengend Snippet: Gemcitabine up-regulates NKG2D ligands in lung cancer cells. (A) qRT-PCR of H60, Raet-1 , and Ulbp1 mRNA in LLC cells. (B) qRT-PCR of major histocompatibility complex class I polypeptide-related sequence A ( MICA ), MICB , and UL16 binding protein ( ULBP ) 1-6 mRNA in A549 cells. Data are the fold-change of mRNA expression in gemcitabine- and cisplatin-treated cells relative to untreated cells. (C) Surface expression of MICA/B, ULBP1, ULBP2/5/6, and ULBP3 was measured by flow cytometry on A549 cells treated with vehicle control (DMSO), gemcitabine or cisplatin. Data are representative of three independent experiments. One-way analysis of variance (ANOVA) was used. * p < 0.05, ** p < 0.01, **** p < 0.0001.

Article Snippet: Anti-human ULBP1, ULBP2/5/6, and ULBP3 antibody were purchased from R&D Systems.

Techniques: Quantitative RT-PCR, Immunopeptidomics, Sequencing, Binding Assay, Expressing, Flow Cytometry, Control

Journal: Journal for Immunotherapy of Cancer

Article Title: CHMP2A regulates broad immune cell-mediated antitumor activity in an immunocompetent in vivo head and neck squamous cell carcinoma model

doi: 10.1136/jitc-2023-007187

Figure Lengend Snippet: EVs secreted by 4MOSC cell lines express mouse NKG2D ligands. (A) 4MOSC1 and 4MOSC2 WT and mCHMP2A KO derived EVs were collected and analyzed by flow cytometry for mouse NKG2D ligands mRae, mULBP, mH60 (blue histogram). Red histogram represents isotype. EVs, extracellular vehicles; KO, knock-out.

Article Snippet: Isolated EVs were incubated with primary staining with mRae (AF1136-SP, R&D Systems) mULBP (MAB2588-SP, R&D Systems), and mH60 (MAB1155-SP, R&D Systems) for 30 min on ice and washed with flow buffer.

Techniques: Derivative Assay, Flow Cytometry, Knock-Out

Journal: Brain : a journal of neurology

Article Title: TGF-beta and metalloproteinases differentially suppress NKG2D ligand surface expression on malignant glioma cells.

doi: 10.1093/brain/awl205

Figure Lengend Snippet: Fig. 1 NKG2DL expression in human gliomas in vivo. Normal white matter (upper left quadrant in each group of four), WHO grade II astrocytoma (upper right), WHO grade III astrocytoma (lower left) and glioblastoma (WHO grade IV) (lower right) were stained with either isotype control [(A), representative IgG1 isotype control is depicted] or specific MICA (B), MICB (C), ULBP1 (D), ULBP2 (E) or ULBP3 (F) antibodies.

Article Snippet: Polyclonal goat anti-human ULBP2 antibody (clone ILU01, R&D Systems) was followed by biotinylated rabbit antigoat immunoglobulins (DakoCytomation).

Techniques: Expressing, In Vivo, Staining, Control

Journal: Brain : a journal of neurology

Article Title: TGF-beta and metalloproteinases differentially suppress NKG2D ligand surface expression on malignant glioma cells.

doi: 10.1093/brain/awl205

Figure Lengend Snippet: Fig. 2 NKG2DL mRNA expression and protein levels at the cell surface: modulation by siRNA targeting TGF-b1/2. (A) MICA, MICB, ULBP1, ULBP2, ULBP3 and ULBP4 mRNA expression was assessed in control or TGF-b1/2 siRNA cells by real-time PCR. (B) MICA, MICB, ULBP1, ULBP2 and ULBP3 protein levels at the cell surface of control (closed profiles) or TGF-b1/2 siRNA cells (open profiles) were assessed by flow cytometry. (C) TGF-b1/2 siRNA cells were untreated or treated with TGF-b2 (10 ng/ml) for 48 h and assessed for MICA, ULBP2 and ULBP4 mRNA expression. (D) TGF-b1/2 siRNA cells, untreated (closed profiles) or treated (open profiles) with TGF-b2 (10 ng/ml) for 7 days, were analysed accordingly. Data in A and C are expressed as the relative gene expression compared with control transfectants in A or untreated TGF-b1/2 siRNA cells in C set to 1. Data represent mean values 6 SEM from three independent experiments (*P < 0.05; **P < 0.01). In B and D SFI values are indicated in the upper right corner and are representative of three independent experiments.

Article Snippet: Polyclonal goat anti-human ULBP2 antibody (clone ILU01, R&D Systems) was followed by biotinylated rabbit antigoat immunoglobulins (DakoCytomation).

Techniques: Expressing, Control, Real-time Polymerase Chain Reaction, Flow Cytometry, Gene Expression

Journal: Brain : a journal of neurology

Article Title: TGF-beta and metalloproteinases differentially suppress NKG2D ligand surface expression on malignant glioma cells.

doi: 10.1093/brain/awl205

Figure Lengend Snippet: Fig. 3 MICA and ULBP2 enhance NK cell lysis of TGF-b1/2 siRNA cells. Control or TGF-b1/2 siRNA cells untreated or treated with anti-MICA (AUMO1), anti-ULBP2 (BUMO1), both antibodies or anti-NKG2D (MAB 139) (10 mg/ml) were used as target cells in a standard 4 h 51Cr release assay using polyclonal NK cells as effectors. Data are expressed as specific lysis at different E : T ratios. One representative experiment of three is shown. Each data point was generated from triplicates. Coefficients of variation are shown as bars (for statistical analysis, see text).

Article Snippet: Polyclonal goat anti-human ULBP2 antibody (clone ILU01, R&D Systems) was followed by biotinylated rabbit antigoat immunoglobulins (DakoCytomation).

Techniques: Lysis, Control, Release Assay, Generated

Journal: Science advances

Article Title: γδ-Enriched CAR-T cell therapy for bone metastatic castrate-resistant prostate cancer.

doi: 10.1126/sciadv.adf0108

Figure Lengend Snippet: Fig. 1. BTN expression and spontaneous gamma/delta T cell infiltration in prostate tumors. (A) Bioinformatic analysis of TCR gamma variable regions (TRGV), BTN3A1, BTN2A1, ULBP1, MICA, MICB, and PSCA in bone and soft tissue biopsies from 118 individuals with mCRPC (SU2C/PCF dataset). (B) Specific comparison of TRGV expression in bone metastases. RPKM, reads per kilobase of transcript per million mapped reads. (C) BTN2A1 and BTN3A1 expression in bone, liver, lymph node (LN), and other soft tissue metastasis sites compared to primary prostate tumors. (D) Representative multiplex staining of bone metastasis sample from TMA containing 1- mm cores. (E) Expression of BTN3A1 and stress markers MICA/B and ULBP1 in tumor cells (PCK+) from a TMA containing samples from patients with metastatic prostate cancer in different tissues. Only samples with more than 100 PCK+ cells were considered for the analysis (n = 64, 27, 52, and 70 for bone, lung, lymph node, and liver samples, respectively). Each dot represents a sample. Bars represent means ± SEM. Linear mixed effect model was used for statistical analysis.

Article Snippet: 9, eadf0108 (2023) 3 May 2023 13 of 18 ow nloaded from https://w w w .science.org on February 27, 2024 dye540), ULBP1 (Proteintech, Rb polyclonal, 1:6000, dye520), εCD3 (DALO, Rb polyclonal, 1:400, dye620), BTN3A1 (Millipore Sigma, Rb polyclonal, 1:50, dye570), and PCK (Abcam, [KRT/ 1877R], 1:150, dye690).

Techniques: Expressing, Comparison, Multiplex Assay, Staining