Journal: Frontiers in Immunology

Article Title: Potent immunomodulatory and antitumor effect of anti-CD20-IL2no-alpha tri-functional immunocytokine for cancer therapy

doi: 10.3389/fimmu.2022.1021828

Figure Lengend Snippet: Anti-huCD20(hγ1)-IL2no-alpha ICK induces NK-cell activation and cytotoxic function. (A–C) , freshly human PBMC (used as a source of NK cells) and Ramos cells were cocultured at a 10:1 ratio in presence of 6nM ICKs (Anti-huCD20(hγ1)-IL2no-alpha or isotype control ICK), or RTX (6nM), or RTX (6nM) + mutein IL-2no-alpha (equimolar amount of cytokine). Flow cytometry analyses of NK cell (CD3-CD56+) activation membrane markers (A) , CD69, (B) CD16A, and degranulation marker (C) CD107a are shown (D–F) , freshly human PBMC and PBMC of refractory low- grade B-cell lymphoma cells were cocultured at a 10:1 ratio for 21 h at 37°C. Flow cytometry analyses of NK cell activation markers (D) , CD16A and (E) degranulation marker CD107a are shown. Percentage is referred to as the proportion of cells within the whole NK cell population expressing CD69, CD16A or CD107a markers. (F) IFN-γ levels in supernatants of cell cultures measured by ELISA. Data represent the mean ± SEM. (Kruskal-Wallis, Dunn’s post hoc test, *P < 0.05; **P <0.01; ***P < 0.001; ns, not significant).

Article Snippet: The following antibodies were used to detect cell surface antigens: anti-CD3-APC (allophycocyanin, BD PharMingen), anti-CD56-FITC (fluorescein isothiocyanate, Becton Dickinson), anti-CD69-PeCy7(phycoerythrin cyanine-7, BD PharMingen), anti-CD16-AF-700 (AlexaFluor 700, Invitrogen) and anti-107a-PE (phycoerythrin, BD PharMingen).

Techniques: Activation Assay, Flow Cytometry, Marker, Expressing, Enzyme-linked Immunosorbent Assay

Journal: Experimental and Therapeutic Medicine

Article Title: Effects of CD4 + T lymphocytes from ovariectomized mice on bone marrow mesenchymal stem cell proliferation and osteogenic differentiation

doi: 10.3892/etm.2020.9212

Figure Lengend Snippet: CD4 + T cells are activated in OVX mice. (A) The gating strategies used for CD4 + T cells. (B) Percentage of CD4 + CD69 + cells in spleens tested using flow cytometry. (C) Expression levels of proinflammatory cytokines, IL-2, IFN-γ and TNF-α, in purified CD4 + T cells were determined using reverse transcription-quantitative-PCR. (D) TNF-α levels in the supernatant from purified CD4 + T cell cultures were measured by ELISA. * P<0.05, ** P<0.01 vs. sham. CD, cluster of differentiation; OVX, ovariectomized; IL-2, interleukin 2; IFN-γ, interferon- γ; TNF-α, tumor necrosis factor-α; IgG, immunoglobulin; APC, allophycocyanin.

Article Snippet: Inc.) and magnetic beads coated with anti-mouse CD4 antibodies (Miltenyi Biotec., Inc.), CD3 and CD28 (both, BD Biosciences), a volumetric microscope, an inverted phase contrast microscope and camera system (Olympus Corporation), a flow cytometer (Beckman Coulter, Inc.), an Alkaline Phosphatase Staining kit (cat. no. P0321; Beyotime Institute of Biotechnology), a mouse TNF-α ELISA kit (cat. no. MTA00B; R&D Systems, Inc.), anti-TNF-α antibodies (cat. no. AF-410-NA; R&D Systems, Inc.), FITC-conjugated anti-CD3 (cat. no. 11-0032-82; eBioscience; Thermo Fisher Scientific, Inc.), allophycocyanin (APC)-conjugated anti-CD4 (cat. no. 17-0042-82; eBioscience; Thermo Fisher Scientific, Inc.), phycoerythrin (PE)-conjugated anti-CD69 (cat. no. 12-0691-82; eBioscience; Thermo Fisher Scientific, Inc.) and Armenian hamster IgG isotype control (cat. no. 12-4888-81; eBioscience; Thermo Fisher Scientific, Inc.).

Techniques: Flow Cytometry, Expressing, Purification, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay