neutralizing antibodies against cytokines  (Thermo Fisher)

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Nanoparticle-mediated combinatorial targeting of multiple human dendritic cell (DC) subsets leads to enhanced T cell activation via IL-15-dependent DC crosstalk

doi: 10.4049/jimmunol.1400489

Figure Lengend Snippet: (6a). The effects of nanoparticles on isolated BDCA3+ myeloid dendritic cells and DC SIGN positive monocyte DCs was studied by incubating them overnight with NP-FMP (peptide concentration 0.05μg/ml) at 37°C. The supernatants were then collected and analyzed by VeriPlex™ Human Cytokine ELISA Kit. BDCA3+ MDCs and DC-SIGN DCs incubated alone were used as negative controls. The graph shows mean cytokine expression levels (IL-15, IFN-lambda, TNFα, IL-6 and IL8) per 30,000 APCs ± standard error of the mean for cells obtained from 3 different healthy donors. * p<0.05

Article Snippet: In additional experiments, neutralizing antibodies against cytokines IL-15, IFN-lambda, IL-6 or isotype control mouse IgG1 (all 10μg/ml, R&D) were added to the combinatorial targeting condition.

Techniques: Isolation, Concentration Assay, Enzyme-linked Immunosorbent Assay, Incubation, Expressing

Journal: Journal of Neuroinflammation

Article Title: CXCR3 signaling in glial cells ameliorates experimental autoimmune encephalomyelitis by restraining the generation of a pro-Th17 cytokine milieu and reducing CNS-infiltrating Th17 cells

doi: 10.1186/s12974-016-0536-4

Figure Lengend Snippet: mRNAs for inflammatory chemokines and cytokines required for Th17 cell expansion are increased in CXCR3 −/− mice during EAE. WT ( n = 5) and CXCR3 −/− ( n = 5) mice were immunized with MOG. Spinal cords were collected on days 0, 10, and 14 post-immunization, and total RNA was extracted and subjected to real-time PCR analysis to determine the mRNA levels for cytokines ( a ) and chemokines ( b ). mRNA levels of target genes, normalized to the level of GAPDH , were analyzed using the ΔΔCt method. n stands for the number of mice. * P < 0.05; ** P < 0.01

Article Snippet: Cells (4 × 10 6 cell/ml) were cultured at 37 °C in a humidified 5 % CO 2 atmosphere for 72 h in RPMI 1640 medium (Life Technologies) supplemented with 10 % FBS (Thermo Scientific, Rockford, IL); 2 mM l -glutamine; 50 μM 2-mercaptoethanol (Merck, Whitehouse Station, NJ); 100 U/ml of penicillin and 100 μg/ml of streptomycin; 10 μg/ml of MOG 35–55 ; Th17-polarizing cytokines, including IL-6 (20 ng/ml; R&D Systems, Minneapolis, MN), IL-23 (20 ng/ml; R&D Systems), and transforming growth factor (TGF)- β (5 ng/ml; PeproTech, Rocky Hill, NJ); and neutralizing antibodies against various cytokines (eBioscience), including IL-4 (10 μg/ml), IFN-γ (10 μg/ml), and IL-2 (10 μg/ml).

Techniques: Real-time Polymerase Chain Reaction