dgka  (Bioss)

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Combination therapy for hepatocellular carcinoma with diacylglycerol kinase alpha inhibition and anti-programmed cell death-1 ligand blockade

doi: 10.1007/s00262-021-03041-z

Figure Lengend Snippet: Inhibition of DGKα suppressed proliferation of liver cancer cells and activated immune cells. a, b Proliferation and viability of human and murine liver cancer cells (n = 4 each) cultured with or without DGKAI were evaluated by Cell Counting kit-8 assay. *One-way ANOVA with Dunnett’s test. c, d IL-2 production in stimulated PBMCs from healthy donors and mouse spleen cells with or without DGKAI was evaluated by ELISA. *One-way ANOVA with Dunnett’s test. e, f Total and phosphorylated ERK and JNK in stimulated mouse spleen cells with and without DGKAI were evaluated by western blotting and densitometry. Mean and SD are presented. OD, optical density

Article Snippet: Sections were washed with Tris-buffered saline and then incubated with anti-human DGKα polyclonal antibodies (11547-1-AP, Proteintech), anti-CD4 mAb (4B12, Leica), or anti-CD8a mAb (C8/144G, Dako) for 30 min, followed by washing with Tris-buffered saline and incubation with Envision FLEX HRP (Dako) at room temperature for 30 min.

Techniques: Inhibition, Cell Culture, Cell Counting, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Combination therapy for hepatocellular carcinoma with diacylglycerol kinase alpha inhibition and anti-programmed cell death-1 ligand blockade

doi: 10.1007/s00262-021-03041-z

Figure Lengend Snippet: Inhibition of DGKα augmented CD8+ effector T cells in liver in tumor-bearing mice. a Liver tumors evaluated by in vivo imaging on day 14. b Photon flux determined from tumor images (n = 7). *One-way ANOVA with Tukey’s test. c HE-stained images of liver tissues from mice on day 14. Bars = 500 μm. d Tumor area calculated from HE-stained sections (n = 4 mice) on day 14. *One-way ANOVA with Tukey’s test. e Infiltration of CD3+ T cells on day 14. Bars = 500 μm. f Number of CD3+ cells in liver tissues based on immunohistochemistry sections (n = 4 mice) on day 14. *One-way ANOVA with Tukey’s test. Mean and SD are presented

Article Snippet: Sections were washed with Tris-buffered saline and then incubated with anti-human DGKα polyclonal antibodies (11547-1-AP, Proteintech), anti-CD4 mAb (4B12, Leica), or anti-CD8a mAb (C8/144G, Dako) for 30 min, followed by washing with Tris-buffered saline and incubation with Envision FLEX HRP (Dako) at room temperature for 30 min.

Techniques: Inhibition, In Vivo Imaging, Staining, Immunohistochemistry

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: Combination therapy for hepatocellular carcinoma with diacylglycerol kinase alpha inhibition and anti-programmed cell death-1 ligand blockade

doi: 10.1007/s00262-021-03041-z

Figure Lengend Snippet: DGKα inhibition augmented anti-tumor effect of PD-L1 in tumor-bearing mice. a Surface expression of PD-L1 on Hepa1-6 cells stimulated with DGKAI or IFN-γ evaluated by flow cytometry. b ∆MFIs compared with isotype controls (n = 3). *One-way ANOVA with Dunnett’s test. c PD-L1 expression in model mice. Bars = 500 μm. d PD-L1+ cells in liver sections (n = 4 mice). e In vivo imaging of liver tumors in mice injected with IgG or anti-PD-L1 mAb. f Photon flux determined from tumor images (n = 4). g HE-stained liver tissues from model mice. Bars = 500 μm. h Tumor area based on HE-stained sections (n = 4 mice). *One-way ANOVA with Tukey’s test. Mean and SD presented

Article Snippet: Sections were washed with Tris-buffered saline and then incubated with anti-human DGKα polyclonal antibodies (11547-1-AP, Proteintech), anti-CD4 mAb (4B12, Leica), or anti-CD8a mAb (C8/144G, Dako) for 30 min, followed by washing with Tris-buffered saline and incubation with Envision FLEX HRP (Dako) at room temperature for 30 min.

Techniques: Inhibition, Expressing, Flow Cytometry, In Vivo Imaging, Injection, Staining

Journal: Frontiers in Immunology

Article Title: Loss of Diacylglycerol Kinase α Enhances Macrophage Responsiveness

doi: 10.3389/fimmu.2021.722469

Figure Lengend Snippet: Increased expression of macrophage activation markers in Dgka -/- BMDM following LPS or IL4 stimulation. (A) WT and Dgka -/- BMDMs were treated with 500ng/mL LPS or 2ng/mL IL-4 for 24h and assessed by qPCR for changes in Nos2 and Arg1 expression, respectively. (B) Immunoblot depicting protein expression of iNOS in lysates of LPS treated WT and Dgka -/- BMDMs (left) and arginase expression of IL-4 treated BMDMs. Quantification of band intensity normalized to PBS treated WT mice (not shown) are indicated in text below bands. (C) Summary plot depicting quantification of immunoblot band intensity of two independent experiments as in (B) for n = 6 per group. (D) WT and Dgka -/- BMDMs treated with LPS and IL-4 were assessed by qPCR for expression of SOCS1 and SOCS3. In (A–D) samples were normalized to the PBS treated WT control. (E) immunoblot of WT and Dgka -/- BMDM whole cell lysates with anti-DGKα antibody. Each data point represents an individual mouse for n = 3 per group. Error bars: SD. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001; ns, not significant; Two-way ANOVA with Tukey post-hoc test.

Article Snippet: Primary antibodies used were specific to F4/80 (Novus Biologicals, #NB800-404), Iba1 (Novus Biologicals, #NB100-1028), and DGKα (Bioss Antibodies, #BS-14294R).

Techniques: Expressing, Activation Assay, Western Blot