Journal: Oncology Letters

Article Title: Role of programmed death ligands in effective T-cell interactions in extranodal natural killer/T-cell lymphoma

doi: 10.3892/ol.2014.2356

Figure Lengend Snippet: Representative immunohistochemical streptavidin-peroxidase staining in extranodal natural killer/T-cell lymphoma (upper row) and rhinitis tissues (lower row). The positive cases of (A and D) programmed death 1, (B and E) PD-L1 and (C and F) PD-L2 (magnification, ×200). PD-L, programmed death ligand.

Article Snippet: The antigen retrieval was conducted in 0.01 mol/l citrate (pH 6.0) and the slides were incubated overnight with rabbit anti-human PD-L1 polyclonal antibody (1:120; Proteintech, Chicago, IL, USA), rabbit anti-human PD-L2 polyclonal antibody (1:150) and mouse anti-human PD-1 monoclonal antibody (mAb; 1:100) (both ZSGB-BIO, Beijing, China) and phosphate-buffered saline was used as a blank control.

Techniques: Immunohistochemical staining, Staining

Journal: Oncology Letters

Article Title: Role of programmed death ligands in effective T-cell interactions in extranodal natural killer/T-cell lymphoma

doi: 10.3892/ol.2014.2356

Figure Lengend Snippet: PD-1 expression in (A) CD4 + and (B) CD8 + T-cell subsets in 20 ENKL patients was significantly increased compared with that in 10 HVs (P<0.05). Representative PD-1 expression in (C) CD4 + and (D) CD8 + T-cell subsets in six ENKL patients was (E) downregulated with chemotherapy. (F) T-helper cell type 1 cytokine (IL-2 and IFN-γ) mean production levels in the serum of 20 ENKL patients were significantly lower than those in 10 HVs (P<0.05). PD1, programmed death 1; ENKL, extranodal natural killer/T-cell lymphoma; HVs, healthy volunteers; IL-2 interleukin 2; IFN-γ, interferon γ.

Article Snippet: The antigen retrieval was conducted in 0.01 mol/l citrate (pH 6.0) and the slides were incubated overnight with rabbit anti-human PD-L1 polyclonal antibody (1:120; Proteintech, Chicago, IL, USA), rabbit anti-human PD-L2 polyclonal antibody (1:150) and mouse anti-human PD-1 monoclonal antibody (mAb; 1:100) (both ZSGB-BIO, Beijing, China) and phosphate-buffered saline was used as a blank control.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Role of programmed death ligands in effective T-cell interactions in extranodal natural killer/T-cell lymphoma

doi: 10.3892/ol.2014.2356

Figure Lengend Snippet: (A) Purity of CD8 + T cells separated by magnetic-activated cell sorting was 99%. (B) Purity of CD8 + PD-1 + T cells was 96.2% following the stimulation of allogeneic CD8 + T cells with phytohemagglutinin for 48 h. (C) SNK-6 cells were used as the control group and, following the coculture of SNK-6 cells and CD8 + T cells for 72 h, a significant inhibitory effect of PD-L1 on allogeneic CD8 + T-helper type 1 cytokine (IL-2 and IFN-γ) secretion was observed; (A and B) P<0.05. (D) CD8 + T-cell apoptosis in groups A and B was not altered significantly compared with activated CD8 + T cells at 72 h (P>0.05). (E) SNK-6 cells were used as the control group and cells harvested at 0, 24, 48 and 72 h were analyzed by flow cytometry gating CFSE + events. The proliferation index was not significantly different among the groups (P>0.05). PD-1, programme death 1; PD-L. programmed death ligand; IL-2 interleukin 2; IFN-γ, interferon γ; CFSE, carboxy-fluorescein succinimidyl ester.

Article Snippet: The antigen retrieval was conducted in 0.01 mol/l citrate (pH 6.0) and the slides were incubated overnight with rabbit anti-human PD-L1 polyclonal antibody (1:120; Proteintech, Chicago, IL, USA), rabbit anti-human PD-L2 polyclonal antibody (1:150) and mouse anti-human PD-1 monoclonal antibody (mAb; 1:100) (both ZSGB-BIO, Beijing, China) and phosphate-buffered saline was used as a blank control.

Techniques: FACS, Flow Cytometry

Journal: Cell Reports Medicine

Article Title: Discovery of galectin-8 as an LILRB4 ligand driving M-MDSCs defines a class of antibodies to fight solid tumors

doi: 10.1016/j.xcrm.2023.101374

Figure Lengend Snippet:

Article Snippet: The plate was then washed with PBST (PBS, 0.05% Tween 20) (MA0015, Meilunbio) (T8220, Solarbio) and blocked with 3% BSA (97061-420, VWR) at 37°C for 90 min. After repeated washing, hFc-tagged recombinant proteins, including CD3ε (10977-H02H; Sino Biological), CTLA-4 (CT4-H5255; Acro Biosystems), CD28 (CD8-H525a; Acro Biosystems), CD96 (TAE-H5252; Acro Biosystems), LAG-3 (LA3-H5255; Acro Biosystems), TIM-3 (TM3-H5258; Acro Biosystems), CD40 (CD0-5253; Acro Biosystems), ICOS (ICS-H5258; Acro biosystems), OX40 (OX0-H5255; Acro Biosystems), TIGHT (TIT-H5254; Acro Biosystems), LY86 (10242-H02H; Sino Biological), LILRB4 (16742-H02H; Sino Biological), CD27 (CD7-H5254; Acro biosystems), PD-1 (10377-H02H; Sino Biological), and CD8b (11031-HCCH; Sino Biological), were added into each well.

Techniques: Recombinant, Derivative Assay, Microarray, Enzyme-linked Immunosorbent Assay, Lysis, Blocking Assay, Staining, Transfection, Marker, Flow Cytometry, shRNA, Plasmid Preparation, Negative Control, Silver Staining, Bicinchoninic Acid Protein Assay, Selection, Extraction, Labeling, RNA Expression, Cell Culture, Software

Journal: Molecular Therapy Oncolytics

Article Title: A PD-L1-Based Cancer Vaccine Elicits Antitumor Immunity in a Mouse Melanoma Model

doi: 10.1016/j.omto.2019.06.002

Figure Lengend Snippet: Antibody Responses Induced by DPDL1E Vaccination in C57BL/6 and BALB/c Mice (A) Mice (n = 8) were immunized with DPDL1E three times at 2-week intervals. One week after the third immunization, the antibody titers were measured by ELISA using His-tagged PDL1 recombinant protein as a coating antigen. DTT-immunized serum was used as a negative control. (B) The levels of PD-L1- and DTT-specific antibody subclasses induced by DPDL1E vaccination. Sera were isolated from C57BL/6 and BALB/C mice immunized with the DPDL1E vaccine. The levels of the indicated antibody subclasses were measured by ELISA. (C) The sera from DPDL1E-immunized mice inhibited binding of PD-L1 to PD-1. PD-L1 mAb at 20 μg/mL was used as a positive control, and sera from DTT-immunized mice and PBS were used as a negative control and blank control, respectively. (D) The inhibition efficiency of sera at different concentrations was tested and compared with the control group. (E) A standard curve was created (relative inhibition versus concentration of PD-L1 mAb) to calculate the effective anti-PD-L1 concentration in vitro . ****p < 0.0001.

Article Snippet: The plates were incubated at room temperature for 1 h. After washing, 100 μL (2.5 μg/mL) biotin-labeled PD-1 protein (Acrobiosystems, Bethesda, MD, USA) was added to the wells and incubated for 1 h at 37°C.

Techniques: Enzyme-linked Immunosorbent Assay, Recombinant, Negative Control, Isolation, Binding Assay, Positive Control, Inhibition, Concentration Assay, In Vitro

Journal: Molecular Therapy Oncolytics

Article Title: A PD-L1-Based Cancer Vaccine Elicits Antitumor Immunity in a Mouse Melanoma Model

doi: 10.1016/j.omto.2019.06.002

Figure Lengend Snippet: The DPDL1E Vaccine Decreased the Ratio of PD-1+ Cells and Increased the Ratio of PD-L1+ Cells in Tumors (A) PD-L1+ cells were stained with anti-CD274-APC and anti-CD45-FITC. (B) PD-1+ cells were stained with anti-CD279-APC and anti-CD45-FITC. (C) LAG3+ cells were stained with anti-CD223-PE, anti-CD3-FITC, and anti-CD8-PerCP-Cy5.5. (D) LAG3+PD-1+ cells were stained with anti-CD279-APC, anti-CD223-PE, anti-CD3-FITC, and anti-CD8-PerCP-Cy5.5. All samples were detected using FCM, and the data were analyzed using FlowJo software. *p < 0.05, **p < 0.01, ***p < 0.001. Statistically significant differences were assessed using Student’s t test.

Article Snippet: The plates were incubated at room temperature for 1 h. After washing, 100 μL (2.5 μg/mL) biotin-labeled PD-1 protein (Acrobiosystems, Bethesda, MD, USA) was added to the wells and incubated for 1 h at 37°C.

Techniques: Staining, Software