Human (#407306) and mouse (#407320) IFN-γ, human IL-6 (#407652) were purchased from Merck Millipore, human IFN-β (#11415–1) was from PBL Assay Science. Human IL-6Rα was from R&D Systems (#227-58-025), human IL-27 from Peprotech (#200–38). Other treatments used in this work include LPS (#L7895, Sigma) and ratjadone (#553590, Merck Millipore). Unless stated otherwise, IFN treatment was for one hour with 50 U/ml IFN-γ or 500 U/ml IFN-β in growth medium. Similarly, unless stated otherwise, IL-27 treatment was performed for 40 min at a concentration of 100 ng/ml. IL-6/IL-6R cotreatments also lasted for 40 min at concentrations of 200 and 250 ng/ml, respectively.
Human il 27
Manufactured by Thermo Fisher Scientific
Sourced in United States
Human IL-27 is a recombinant protein that represents the cytokine interleukin-27. It is a member of the interleukin-6 cytokine family and plays a role in regulating immune responses.
Automatically generated - may contain errors
Lab products found in correlation
Human ifn β, by PBL Assay Science (1 mentions)
Human il 6rα, by R&D Systems (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Fludarabine, by Selleck Chemicals (1 mentions)
Stattic, by Selleck Chemicals (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Hl 60 cells, by American Type Culture Collection (1 mentions)
4 protocols using human il 27
1
Cytokine signaling in immune cells
2
Regulation of CD8+ T Cells by IL-27
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CD8+ T cells were cultured in the CM with different concentrations of human IL-27 (PeproTech) or not (control). For anti-IL-27 blocking experiments, CD8+ T cells were stimulated with supernatant and 10 μg/mL anti-IL-27 antibody (R&D Systems) or IgG isotype (control). Similarly, CD8+ T cells were stimulated with 50 ng/mL IL-27, 62.5 nM STAT-3′ inhibitor Stattic (Selleck), or 100 μM STAT1 inhibitor Fludarabine (Selleck) for STAT1/3 blocking tests. The cells mentioned above were dealt with 48 h then subjected to further analysis.
3
Investigating IL-27's Role in Tr1 Cells in IgAV
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To determine the role of IL-27 on Tr1 cells in IgAV, PBMCs from IgAV patients were stimulated with anti-CD3 (2 μg/ml) and anti-CD28 (1 μg/ml) in 96-well plates at a density of 2–5 × 105 cells per well (200 μl). Thereafter, cells were cultured in RPMI 1640 medium (Gibco-Invitrogen, Carlsbad, CA, USA) with 10% fetal bovine serum (Hyclone, South Logan, UT, USA) supplemented with 2 mM l -glutamine, 0.05 mM β-mercaptoethanol, 100 U/ml of penicillin and 100 μg/ml streptomycin at 37°C in 5% CO2 for 5 days in the presence or absence of human IL-27 (100 ng/ml, PeproTech, Rocky Hill, NJ, USA).Then, the cells were collected for flow cytometry and real-time quantitative PCR. Cell culture supernatant was collected for and stored at –20°C until bulk analysis.
4
Chemotaxis Assay for Granulocyte Migration
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HL-60 cells (ATCC) were differentiated into granulocytes using 100 mmol·L−1 dimethylformamide (DMF) (Millipore Sigma) and plated in the top compartment of Boyden chambers, and a chemotaxis assay was performed according to the manufacturer’s protocol (Millipore Sigma QCMTM Chemotaxis 5 μm 24-Well Cell Migration Assay Kit). Briefly, the chemotaxis of 1 × 106 cells per chamber toward the bottom compartment containing RPMI 1640 medium with or without a chemoattractant [human IL-27 (500 ng·mL−1, PeproTech)] or with N-formyl-methionyl-leucyl-phenylalanine (fMLP; 800 ng·mL−1, Millipore Sigma) as a positive control was evaluated by incubation for 1 h at 37 °C. After incubation, cell migration into the bottom chamber was quantified as relative fluorescence units (RFUs) according to the manufacturer’s instructions.
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