proteins proteome profiler human cytokine array kit  (R&D Systems)

Journal: Frontiers in Immunology

Article Title: E2F1-induced autocrine IL-6 inflammatory loop mediates cancer-immune crosstalk that predicts T cell phenotype switching and therapeutic responsiveness

doi: 10.3389/fimmu.2024.1470368

Figure Lengend Snippet: The impact of E2F1 on immune cell infiltration and cytokine release in coculture with T cells. (A, B) Correlation of E2F1 expression with infiltration of monocytes, eosinophils, and neutrophils in melanoma patients with primary tumors (A) or metastasis (B) . (C, D) Estimation of CD4 + Th1 and Th2 cell infiltration cells in melanoma patients with primary tumors or metastasis correlated to the expression of E2F1, IL-6 (C) , and STAT3, CD274 (D) . Significant correlations are highlighted in red. (E, F) Cytokine assays of supernatants from different culture/coculture conditions were compared as illustrated showing an E2F1-dependent increased secretion of immunosuppressive type 2 cytokines (E) that was completely reversed after E2F1-KD in melanoma cells (F) . Asterisks indicate transcriptionally upregulated factors. FC, fold change.

Article Snippet: To measure secreted proteins Proteome Profiler Human Cytokine Array Kit was used according to manufacturer’s instruction (#ARY005B, R&D Systems).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: E2F1-induced autocrine IL-6 inflammatory loop mediates cancer-immune crosstalk that predicts T cell phenotype switching and therapeutic responsiveness

doi: 10.3389/fimmu.2024.1470368

Figure Lengend Snippet: Mathematical model and overview of the melanoma-CD4 + T cell crosstalk with E2F1-STAT3/IL-6/IL6R axis. (A) Mathematical model of the E2F1-STAT3/IL-6 axis including the auto- and paracrine feedback loops from melanoma and immune cells. The heatmap shows that factors related to the IL6R-STAT3-E2F1 pathway are expressed in melanoma and CD4 + T cells prior to activation and crosstalk. (B) Crosstalk simulation illustrating IL-6 expression in melanoma and CD4 + T cells. (C) Output of simulation representing expression levels of E2F1 or STAT3 that are sufficient to trigger IL-6 secretion in melanoma cells. (D) The impact from E2F1 on the melanoma T cell crosstalk where high E2F1 expression leads to an immunosuppressive TME through feedback loops triggered increased release of IL-6 inducing a Th2 shift (left). In contrast, E2F1 depletion in melanoma cells blocks the augmented IL-6 secretion and, in turn, diminishes type-2 cytokine production favoring an anti-tumor Th1 response.

Article Snippet: To measure secreted proteins Proteome Profiler Human Cytokine Array Kit was used according to manufacturer’s instruction (#ARY005B, R&D Systems).

Techniques: Activation Assay, Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Gasdermin-E Mediated Pyroptosis—A Novel Mechanism Regulating Migration, Invasion and Release of Inflammatory Cytokines in Rheumatoid Arthritis Fibroblast-like Synoviocytes

doi: 10.3389/fcell.2021.810635

Figure Lengend Snippet: Alleviation of IL-6 and other inflammatory cytokines release by downregulation of GSDME-mediated RA-FLSs pyroptosis. (A,C) After siRNA transfection and TNF-α + hypoxia treatment, cell culture supernatant was collected and the levels of cytokines and chemokines were measured using Proteome Profiler Human Cytokine Array Kit from R&D systems (A,B) . Supernatant levels of IL-6, IL-8 were further verified by ELISA (n = 3) (C) . (D,E) HE and IHC staining for IL-6 of synovium tissues from RA ( n = 21), OA ( n = 4) and Orth.A ( n = 5) patients (scale bar = 100 μm). RA synovium tissues were futher divide into low GSDME-NT level ( n = 6) and high GSDME-NT level ( n = 15) groups based on matched GSDME-NT level determined by western blotting. Data are shown as mean ± s. e.m and results are representative of three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001 using one-way ANOVA.

Article Snippet: Supernatants of cells with the indicated treatments were collected and centrifuged (12,000 rpm, 5 min), and secretion of cytokines was analyzed by a Protein Profiler Human Cytokine Array Kit (ARY005B, R&D Systems, United States) following the manufacturer’s protocol.

Techniques: Transfection, Cell Culture, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Western Blot