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Rmil 18

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RmIL-18 is a recombinant mouse interleukin-18 protein. Interleukin-18 is a proinflammatory cytokine that plays a role in the immune response.

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Lab products found in correlation

Rmil 15, by Thermo Fisher Scientific (2 mentions) Rhil 2, by Thermo Fisher Scientific (2 mentions) Shp099, by Selleck Chemicals (1 mentions) Rmil 10, by Thermo Fisher Scientific (1 mentions) Rmtgf β, by R&D Systems (1 mentions) Ls column, by Miltenyi Biotec (1 mentions) Anti fitc microbeads, by Miltenyi Biotec (1 mentions) Cytofix cytoperm, by BD (1 mentions)

5 protocols using rmil 18

1

Enhancing Immunotherapy Efficacy in Mice

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Mice were treated once a day with an intraperitoneal injection of 10,000 IU/head rhIL2 (PeproTech) and 1 μg/head rmIL18 (R&D Systems). These cytokines were suspended in 200 μL of sterile PBS, and mice treated with 200 μL PBS were used as controls. Anti-mouse PD-L1 antibody (10F.9G2; BioLegend), anti-mouse PD-L2 antibody (TY25; BioLegend), anti-mouse VEGF antibody (B20–4.1.1; gifted from Genentech) or Rat IgG (MP Biomedicals) was administered intraperitoneally to the mice at a dose of 10 mg/kg three times from the day before cytokine treatment. Wet lung weight and moist rale were measured after 5 days of the treatment.
Iwai T., Sugimoto M., Patel H., Yorozu K., Kurasawa M, & Kondoh O. (2021). Anti-VEGF Antibody Protects against Alveolar Exudate Leakage Caused by Vascular Hyperpermeability, Resulting in Mitigation of Pneumonitis Induced by Immunotherapy. Molecular Cancer Therapeutics, 20(12), 2519-2526.
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2

Modulation of NK Cell Responsiveness

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To study the responsiveness of NK cells to IL-15, splenocytes were resuspended with RPMI 1640 medium and plated on 24-well plates. Then splenocytes were treated with rmIL-15 (Peprotech, 100 ng/mL) for 1, 2, 3, 5, 7, or 9 days, followed by flow cytometry. To investigate the influence of cytokines on METTL3 expression in NK cells, we treated splenocytes with rmIL-17F (25 ng/mL, Peprotech), rmIL-27 (25 ng/mL, Peprotech), rmIL-10 (25 ng/mL, Peprotech), rmIL-15 (25 ng/mL, Peprotech), rmTGF-β (25 ng/mL, R&D) and rmIL-18 (25 ng/mL, R&D) for 3 days. To study the effect of MC38 cells on METTL3 expression in NK cells, purified NK cells were cocultured with MC38 cells under the stimulation of rmIL-15 (10 ng/mL) with the presence or absence of SIS3 (20 μM, APExBIO, Houston, TX, USA) or GW788388 (20 μM, APExBIO) for 2 days. To observe the effects of SHP-2 on the NK cells, splenocytes were treated with rmIL-15 (50 ng/mL, Peprotech) with or without SHP-2 inhibitor SHP099 (4 μM, Selleck, Houston, TX, USA) for 3 days.
Song H., Song J., Cheng M., Zheng M., Wang T., Tian S., Flavell R.A., Zhu S., Li H.B., Ding C., Wei H., Sun R., Peng H, & Tian Z. (2021). METTL3-mediated m6A RNA methylation promotes the anti-tumour immunity of natural killer cells. Nature Communications, 12, 5522.
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3

Mature Thymic Cell Isolation and Stimulation

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Thymic single-cell suspensions were obtained by mechanical dissociation trough a 40 μm cell strainer. Enrichment in mature cells (CD24-) was achieved by negative selection using LS columns (Miltenyi) after staining the cells with anti-CD24-FITC antibody (Invitrogen) and anti-FITC microbeads (Miltenyi). 1 x 106 cells/mL were plated for 36h in complete RPMI 1640 media (10% FCS, 100 U Penicillin/Streptomycin, 10 mM Hepes, 1 mM Sodium Pyruvate, 1X Non-essential Amino Acid, 50 μM β-mercaptoethanol) alone or with addition of 5-OP-RU (1.5 μM, 5-OP-RU synthesised in house) or of rmIL18 (10 ng/mL, R&D).
du Halgouet A., Darbois A., Alkobtawi M., Mestdagh M., Alphonse A., Premel V., Yvorra T., Colombeau L., Rodriguez R., Zaiss D., El Morr Y., Bugaut H., Legoux F., Perrin L., Aractingi S., Golub R., Lantz O, & Salou M. (2023). Role of MR1-driven signals and amphiregulin on the recruitment and repair function of MAIT cells during skin wound healing. Immunity, 56(1), 78-92.e6.
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4

Cytotoxicity Assay with YAC-1 Cells

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YAC-1 cells were labeled with CTV (Thermo Fisher Scientific) as described above. Splenocytes were stimulated with recombinant mouse (rm) IL-12 (20 ng/mL, Peprotech), rmIL-15 (20 ng/mL, Peprotech) and rmIL-18 (10 ng/mL, R&D, Minneapolis, MN, USA) for 18 h. Then, the stimulated splenocytes (5 × 106) were cocultured with CTV-labeled YAC-1 cells (5 × 104) for 4 h, followed by an analysis of the BD flow cytometer.
Song H., Song J., Cheng M., Zheng M., Wang T., Tian S., Flavell R.A., Zhu S., Li H.B., Ding C., Wei H., Sun R., Peng H, & Tian Z. (2021). METTL3-mediated m6A RNA methylation promotes the anti-tumour immunity of natural killer cells. Nature Communications, 12, 5522.
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5

Activation and Proliferation of Murine Immune Cells

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Splenocytes were resuspended in RPMI-1640 supplemented with 10% FCS, 50 μM β-mercaptoethanol, 100 μM non-essential amino acids, 1 mM sodium pyruvate, 50 mM HEPES, and 2 μM L-glutamine. One million cells were stimulated for 6 hr in the presence of BD GolgiStop with 20 ng/ml recombinant mouse (rm) IL-12 (R&D Systems #419-ML/CF) and 10 ng/ml rmIL-18 (R&D Systems #B0025), with 20 ng/ml PMA and 200 ng/ml ionomycin, or with 10 μg/ml of plate-bound anti-NK1.1 antibody (clone PK136), were fixed and permeabilized with BD Cytofix/Cytoperm, and then stained for IFN-γ and CD107a. For phosphoSTAT5 (BD #612599) and phosphoSTAT3 (BD #557814) staining, one million cells were stimulated for the indicated times and concentrations with recombinant human (rh) IL-15 (R&D Systems #247-ILB) or rmIL-21 (Zymogenetics), fixed with 1% paraformaldehyde (PFA) in PBS, permeabilized with methanol, and stained. For proliferation assays, one million cell trace violet (Invitrogen #C34557)-labeled splenocytes were cultured in 10 ng/ml rhIL-15, 1×105 PFA-fixed RMA lymphoma cells, or mouse cytomegalovirus m157-transduced RMA cells supplemented with 50 U/ml rhIL-2 (NCI Preclinical Repository) or 100 ng/ml rmIL-21.
Lam V.C., Folkersen L., Aguilar O.A, & Lanier L.L. (2019). KLF12 regulates mouse Natural Killer cell proliferation. Journal of immunology (Baltimore, Md. : 1950), 203(4), 981-989.
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