Journal: Cell reports

Article Title: Chronic stress primes innate immune responses in mice and humans

doi: 10.1016/j.celrep.2021.109595

Figure Lengend Snippet:

Article Snippet: TNF-α, IL-6, and IL-1β levels were measured in the cell supernatant using the Cytometric Bead Array (CBA) multiplex immunoassay (BD Bioscience).

Techniques: Recombinant, Phagocytosis Assay, Isolation, Enzyme-linked Immunosorbent Assay, Software

Journal: Nature Communications

Article Title: Pharmacologic inhibition of protein phosphatase-2A achieves durable immune-mediated antitumor activity when combined with PD-1 blockade

doi: 10.1038/s41467-018-04425-z

Figure Lengend Snippet: LB-100 impairs induction of naive CD4+ T cells into Treg or Th2 lineage. Naive CD4+ T cells were isolated from mice splenocytes and stimulated with anti-CD3 (10 μg ml −1 ) and soluble anti-CD28 (2 μg ml −1 ) under different skewing conditions. After 3 days in culture, intracellular protein expression was analyzed by flow cytometry and cytokines secretion was measured by bead-based multianalyte flow immunoassays. LB-100 was replenished daily with the indicated concentration. a Naive CD4+ cells were induced into Tregs in the presence of TGF-β with titration of LB-100 for 3 days. Intracellular Foxp3 was labeled with flow cytometry. Quantification of CD4+ Foxp3+Treg cells and representative flow cytometry data demonstrated a decrease in % of Foxp3+ CD4+ cells with LB-100. Cells were gated on CD4+ cells. b , c Naive CD4+ cells were induced into Th2 CD4+ cells in the presence of IL4 with titration of LB-100 for 3 days. b Intracellular GATA3 was labeled with flow cytometry. Quantification of CD4+ GATA3+ Th2 cells and representative flow cytometry data demonstrated a decrease in % of GATA3+ CD4+ cells with LB-100. Cells were gated on CD4+ cells. c Ratio of GATA3+ Th2 over T-bet+ Th1 CD4+ cells under Th2 condition was decreased with LB-100. d Purified naive CD4+ T cells were induced in the presence of Th1 or Th2 skewing condition. On day 3, cells were stimulated and intracellular production of IFN-γ was measured by flow cytometry. Quantification of CD4+ IFN-γ+ cells and representative flow cytometry data demonstrated an increase in % of IFN-γ+ CD4+ cells with LB-100 in both Th1 and Th2 conditions. Cells were gated on CD4+ cells. e TNF, IL2, and IFN-γ production in supernatant of naive CD4+ T cells activated in Th1 skewing conditions for 3 days. f TNF, IL2, IFN-γ, and IL4 production in supernatant of naive CD4+ T cells activated in Th2 skewing conditions for 3 days. Cytokine levels were adjusted to absolute cell number. * P < 0.05, ** P < 0.01, *** P < 0.001 (one-way ANOVA with Tukey’s multiple comparison test). Data are from one experiment representative of two independent experiments with similar results. Error bars depict SEM

Article Snippet: Bead-based multianalyte flow immunoassays (BD Bioscience) were used to measure cytokine production in the supernatant per manufacturer’s instruction.

Techniques: Isolation, Expressing, Flow Cytometry, Concentration Assay, Titration, Labeling, Purification