Journal: Frontiers in Immunology

Article Title: Macrophages Control the Bioavailability of Vitamin D and Vitamin D-Regulated T Cell Responses

doi: 10.3389/fimmu.2021.722806

Figure Lengend Snippet: Co-cultures of M1 macrophages and T cells produce sufficient 1,25(OH) 2 D 3 to affect T cell effector function in the presence of high concentrations of DBP. Relative (A) IFN-γ, (B) IL-5 and (C) IL-13 concentrations in the supernatant of CD4 + T cells activated in mono-cultures with Dynabeads Human T-activator CD3/CD28 and CD4 + T cells co-cultured with allogeneic M1 and M2 macrophages activated with LPS and IFNγ and LPS, respectively, for 96 hours in the presence of 100 nM 25(OH)D 3 and DBP at the indicated concentrations. Each series of data was normalized to the cytokine production in the presence of 100 nM 25(OH)D 3 and absence of DBP. Relative expression of Tbx21 (D) , GATA3 (E) , FoxP3 (F) and RORC (G) in CD4 + T cells co-cultured with allogeneic M1 and M2 macrophages activated with LPS and IFNγ and LPS, respectively, for 96 hours in the presence of 100 nM 25(OH)D 3 and DBP at the indicated concentrations. Data (mean ± SEM) were obtained from two to five independent experiments with 6 donors (4 T cells donors, 8 donor pairs for T cells in co-culture with M1 macrophages (4 T cell donors and 2 M1 macrophage donors mixed to 8 combinations) and 6 donor pairs for T cells in co-culture with M2 macrophages (3 T cell donors and 2 M2 macrophage donors mixed to 6 combinations). The data sets were tested using a one-way ANOVA with post hoc multiple comparisons test (Dunnett’s) to the cell cultures without DBP. *T cells treated with DBP versus T cells without DBP; # T cells + M2 macrophages treated with DBP versus T cells + M2 macrophages without DBP; ¤ T cells + M1 macrophages treated with DBP versus T cells + M1 macrophages without DBP.

Article Snippet: IFNγ, IL-4, -5, -10, -13 and -17A were measured by ELISA according to the manufacturer´s instruction (IFNγ (Cat: 88-7316-88), IL-4 (Cat: 88-7046-88), IL-5 (Cat: 88-7056-88), IL-13 Cat: 88-7439-88 and IL-17A (Cat: 88-7176-88) all from ThermoFisher Scientific).

Techniques: Cell Culture, Expressing, Co-Culture Assay

Journal: Redox Biology

Article Title: Molecular mechanism of interleukin-17A regulating airway epithelial cell ferroptosis based on allergic asthma airway inflammation

doi: 10.1016/j.redox.2023.102970

Figure Lengend Snippet: IL-17A knockout suppresses asthma-induced lung inflammation. (A) Sketch map showing the construction of the OVA-mediated asthma model used in this study. (B) HE staining (magnification: 200 × , bar = 100 μm) to observe infiltration of perivascular and peribronchial inflammatory cells and PAS staining (magnification: 400 × , bar = 50 μm) to observe mucus production in the epithelial layer in Sham, OVA, IL-17A−/− sham, and IL-17A−/− OVA groups. (D) The mouse serum in the sham and OVA groups. (E) The mouse serum IgE, IL-4, IL-5, IL-13, and IL-17A levels in the four groups. All experiments were repeated at least three times.

Article Snippet: Serum IL-5 (Cat #: 88–7056, Invitrogen, USA), IL-13 (Cat #: 88–7439, Invitrogen, USA), and IL-17A (Cat #: 88–7176, Invitrogen, USA) levels in cases and serum IgE (Cat #: 88–50460, Invitrogen, USA), IL-4 (Cat #: 88–7044, Invitrogen, USA), IL-5 (Cat #: 88–7054, Invitrogen, USA), IL-13 (Cat #: 88–7137, Invitrogen, USA), along with IL-17A (Cat #: 88–7371, Invitrogen, USA) in mice were detected through enzyme-linked immunosorbent assay (ELISA).

Techniques: Knock-Out, Staining

Journal: Redox Biology

Article Title: Molecular mechanism of interleukin-17A regulating airway epithelial cell ferroptosis based on allergic asthma airway inflammation

doi: 10.1016/j.redox.2023.102970

Figure Lengend Snippet: Ferroptosis and Th17 cell differentiation are related with asthma. (A) KEGG analysis of patients with asthma compared to healthy controls using the GDS4896 dataset. (B) Relative expressions of differential ferroptosis-related genes of patients with asthma compared to healthy controls using the GDS4896 dataset. (C) PPI networks analysis of differential ferroptosis-related genes in patients with asthma compared to healthy controls using the GDS4896 dataset. (D) Differentially expressed genes associated with ferroptosis obtained in GSE4896 were intersected with differential genes taken from GSE198683. (E) IL-17A mRNA levels were increased in SA compared with the healthy controls in the GDS4896 dataset. (F) IL-5, IL-13, and IL-17A levels in serum samples from patients with asthma and healthy children. (G) Fe 2+ , Fe 3+ , Fe 2+ +Fe 3+ , and Fe 2+ /Fe 3+ levels in serum samples from patients with acute asthma attack and healthy children. All experiments were repeated at least three times.

Article Snippet: Serum IL-5 (Cat #: 88–7056, Invitrogen, USA), IL-13 (Cat #: 88–7439, Invitrogen, USA), and IL-17A (Cat #: 88–7176, Invitrogen, USA) levels in cases and serum IgE (Cat #: 88–50460, Invitrogen, USA), IL-4 (Cat #: 88–7044, Invitrogen, USA), IL-5 (Cat #: 88–7054, Invitrogen, USA), IL-13 (Cat #: 88–7137, Invitrogen, USA), along with IL-17A (Cat #: 88–7371, Invitrogen, USA) in mice were detected through enzyme-linked immunosorbent assay (ELISA).

Techniques: Cell Differentiation