Rmil 15

Manufactured by R&D Systems

RmIL-15 is a recombinant mouse Interleukin-15 protein. Interleukin-15 is a cytokine that plays a role in the proliferation and activation of natural killer cells and T cells.

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Lab products found in correlation

Golgistop, by BD (1 mentions) Rmil 12, by R&D Systems (1 mentions) Rmil 15, by Thermo Fisher Scientific (1 mentions) Vivotag s 750, by PerkinElmer (1 mentions) Rmil 2, by R&D Systems (1 mentions) Xenolight dir, by PerkinElmer (1 mentions)

3 protocols using rmil 15

Cytokine-Induced NK Cell Activation

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Lung or spleen single-cell suspensions were prepared and cultured in the presence of GolgiStop (BD) without or with cytokines (25 ng/ml rmIL-12 and 20 ng/ml rmIL-12 from R&D Systems) or on antibody-coated plates (anti-Ly49D at 10 µg/ml) or with B16F10 or YAC1 cell lines (400,000 cells/well) for 4 h at 37°C. Surface and intracellular stainings were then performed, and IFN-γ production was measured by flow cytometry. For phospho flow analyses, spleen cells were stimulated for 1 h in the presence of graded doses of rmIL-15 (R&D Systems) at 37°C. Spleen cell suspensions were also incubated for 24 h at 37°C with graded doses of rmIL-15 from R&D Systems, and NK cell survival was assessed using Annexin V/live-dead staining.

van Helden M.J., Goossens S., Daussy C., Mathieu A.L., Faure F., Marçais A., Vandamme N., Farla N., Mayol K., Viel S., Degouve S., Debien E., Seuntjens E., Conidi A., Chaix J., Mangeot P., de Bernard S., Buffat L., Haigh J.J., Huylebroeck D., Lambrecht B.N., Berx G, & Walzer T. (2015). Terminal NK cell maturation is controlled by concerted actions of T-bet and Zeb2 and is essential for melanoma rejection. The Journal of Experimental Medicine, 212(12), 2015-2025.

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Topical IL-15 Superagonist Treatment for AD

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IL-15 SA was administered by intraperitoneal injection of 1 μg of SA in 100 μl of phosphate-buffered saline (PBS) daily on days 4 to 7 of MC903 treatment. IL-15 SA was prepared as previously described (47 (link)). Briefly, 20 μg of recombinant murine IL-15 (rmIL-15; eBioscience or STEMCELL) was combined with 90 μg of a chimeric sIL-15Rα fused to the Fc domain of human IgG1 (R&D Systems) at a concentration of 0.1 mg/ml of IL-15 in PBS. The mixture was then vortexed, incubated at 37°C for 20 min, and diluted to 10 μg/ml of IL-15 in PBS. SA concentration was calculated with reference to IL-15 for in vivo dosing. Stable loading was confirmed by measuring free rmIL-15 in solution after coincubation of rmIL-15 and IL-15Rα–Fc proteins by enzyme-linked immunosorbent assay (ELISA) (R&D Systems, DuoSet). Actual values based on a standard curve were comparable to values predicted based on molar ratios. Isotype control solution was prepared in the same fashion with rhIgG1 (R&D Systems), 0.1% bovine serum albumin, and 1 μM glycine in PBS. Aliquots of SA or isotype solution were frozen at −20°C and thawed just before injection. Clinical scoring was adapted from the eczema area and severity index (EASI) (70 (link)), performed by a treatment-blinded investigator, and calculated as the sum of a redness score (0 = none, 5 = severe) and a scaling score (0 = none, 5 = severe).

Mack M.R., Brestoff J.R., Berrien-Elliott M.M., Trier A.M., Yang T.L., McCullen M., Collins P.L., Niu H., Bodet N.D., Wagner J.A., Park E., Xu A.Z., Wang F., Chibnall R., Council M.L., Heffington C., Kreisel F., Margolis D.J., Sheinbein D., Lovato P., Vivier E., Cella M., Colonna M., Yokoyama W.M., Oltz E.M., Fehniger T.A, & Kim B.S. (2020). Blood natural killer cell deficiency reveals an immunotherapy strategy for atopic dermatitis. Science translational medicine, 12(532), eaay1005.

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Labeling T cells for in vivo tracking

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Single T cell suspensions derived from erythrocyte-depleted spleen cell suspensions of OT-1 and B6 mice (155 mM NH₄Cl, 10 mM KHCO₃, 0,1 mM EDTA) by activation of 2 × 10⁶ cells/ml splenocytes in T cell medium for 48 hours. OT-1 T cell suspensions were supplemented with 2 µg/ml 50 U/ml rm IL-2 (R&D) and 0.1 µg/ml CD28 (R&D), B6 T cell were activated in CD3 coated cell culture flasks (1 µg/ml, R&D) supplemented with 50 U/ml rmIL-2 and 0.1 µg/ml CD28. Next, cells were expanded at a density of 1 × 10⁶ cells/ml in T cell medium supplemented with 40 ng/ml rmIL-15 (R&D) for additional 3 days. On day 5 after isolation, cells were labeled with 10 µM Xenolight DiR (Perkin Elmer) in 1x PBS at a concentration of 1 × 10⁶ cells/ml for 15 minutes at 37 °C, with 24 µM VivoTag-S 750 (Perkin Elmer) in T cell medium at a concentration of 4 × 10⁶ cells/ml for 30 minutes at 37 °C or double labeled with both dyes. After washing twice with 1x PBS, labeled T cells were incubated for 2 hours in T cell medium with 40 ng/ml IL-15 to recover. Subsequently, labeled cells were transferred into animals or further analysed in vitro. Labeling conditions for VivoTag-750 were analysed using full medium and serum-free medium and the same conditions as described earlier.

Kimm M.A., Tzoumas S., Glasl S., Omar M., Symvoulidis P., Olefir I., Rummeny E.J., Meier R, & Ntziachristos V. (2020). Longitudinal imaging of T cell-based immunotherapy with multi-spectral, multi-scale optoacoustic tomography. Scientific Reports, 10, 4903.

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