human recombinant il-37 protein  (PeproTech)

Journal: Cells

Article Title: Expression of IL-37 Induces a Regulatory T-Cell-like Phenotype and Function in Jurkat Cells

doi: 10.3390/cells11162565

Figure Lengend Snippet: IL37 overexpression induces FOXP3 in Jurkat cells. ( A ) pLenti-IL37-C-Myc-DDK-P2A-Puro vector ( IL37 OE) (Origene). ( B ) qRT-PCR analysis of IL37 mRNA expression in Jurkat cells transfected with an empty vector or IL37 OE. GAPDH served as an internal control, and the values represent IL37 gene expression as a ratio to GAPDH expression. ( C ) IL-37 and FOXP3 immunoblotting of empty vector and IL37 OE Jurkat cells. Actin was used as a loading control. Representative immunoblotting (left panels) and quantification of bands (right panels) of three immunoblot experiments. The band densities of proteins were quantified with Image J, normalized to actin, and expressed as fold changes compared to control Jurkat cells expressing an empty vector. ( D ) FOXP3 mRNA expression in vector control and IL37 OE Jurkat cells. GAPDH served as an internal control, and the values represent FOXP3 gene expression as a ratio to GAPDH expression. ( E ) Flow cytometry analyses of FOXP3 protein expression in vector control and IL37 OE Jurkat cells. Histograms (top) and quantification by mean fluorescent intensity (MFI) (bottom) of FOXP3 expression. IgG used for negative control staining (solid gray histogram). Small horizontal lines indicate mean ± s.e.m. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.001 (Student’s t test). Data are representative of three independent experiments.

Article Snippet: As controls, 1 and 2 ng/mL recombinant human IL-37 (R&D) were used to measure the sensitivity of the ELISA assay.

Techniques: Over Expression, Plasmid Preparation, Quantitative RT-PCR, Expressing, Transfection, Western Blot, Flow Cytometry, Negative Control, Staining

Journal: Cells

Article Title: Expression of IL-37 Induces a Regulatory T-Cell-like Phenotype and Function in Jurkat Cells

doi: 10.3390/cells11162565

Figure Lengend Snippet: IL-37 overexpressing Jurkat cells are phenotypically similar to Treg cells. ( A ) qRT-PCR analysis of CTLA4, IL10, GITR , and TGFB mRNA expression in vector control and IL37 OE Jurkat cells. GAPDH served as an internal control, and the values represent the gene expression level as a ratio to GAPDH expression. ( B ) CTLA-4 protein expression in vector control and IL37 OE Jurkat cells. Histograms (upper) and quantification by mean fluorescent intensity (MFI) (lower) of CTLA-4 expression. IgG was used for negative control staining (solid gray histogram). ( C ) Left, contour plots of IL-10+ cell gating in unstimulated and anti-CD3/CD28 treated vector control and IL37 OE Jurkat cells. Right lower, quantification of the percentage of IL-10+ Jurkat cells. IgG was used for gating for positive staining. Small horizontal lines indicate mean ± s.e.m. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 (Student’s t test). Data are representative of four independent experiments.

Article Snippet: As controls, 1 and 2 ng/mL recombinant human IL-37 (R&D) were used to measure the sensitivity of the ELISA assay.

Techniques: Quantitative RT-PCR, Expressing, Plasmid Preparation, Negative Control, Staining

Journal: Cells

Article Title: Expression of IL-37 Induces a Regulatory T-Cell-like Phenotype and Function in Jurkat Cells

doi: 10.3390/cells11162565

Figure Lengend Snippet: IL-37 overexpressing Jurkat cells are highly suppressive. In vitro T cell suppression assay by vector control and IL37 OE Jurkat cells. Histograms (left) and % proliferation (right) showing division of CFSE-labeled CD4+CD25- T cell responders (Tresp) purified from healthy human donor PBMCs, cultured with anti-CD3 and either vector control Jurkat cells (black bars), IL37 OE Jurkat cells (gray bars), or human primary Treg cells (white bars) at Treg:Tresp ratios of 4:1 to 1:2 for 5 days. Purified Tresp cells without Jurkat cells or Treg cells (0:1) were used as a positive control of Tresp proliferation without suppression (an orange bar). Data represent mean ± s.e.m. NS, not significant ( p > 0.05); ** p < 0.01; *** p < 0.001; **** p < 0.0001 (Student’s t test). Data are representative of two independent experiments.

Article Snippet: As controls, 1 and 2 ng/mL recombinant human IL-37 (R&D) were used to measure the sensitivity of the ELISA assay.

Techniques: In Vitro, Suppression Assay, Plasmid Preparation, Labeling, Purification, Cell Culture, Positive Control