Mouse il 15

Manufactured by Thermo Fisher Scientific

Mouse IL-15 is a recombinant cytokine that belongs to the common gamma chain cytokine family. It is involved in the proliferation and activation of T cells, natural killer cells, and other immune cells. The product is intended for research use only and not for diagnostic or therapeutic purposes.

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Lab products found in correlation

Mouse il 7, by Thermo Fisher Scientific (2 mentions) Recombinant human il 2, by Roche (2 mentions) Rapamycin, by Bio-Techne (2 mentions) S6k1 inhibitor pf 4708671, by Bio-Techne (2 mentions) Recombinant human il 2, by Thermo Fisher Scientific (2 mentions) Brefeldin a, by Merck Group (2 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Facsmelody, by BD (1 mentions) Bz x710 microscope, by Keyence (1 mentions) Rtca software, by Agilent Technologies (1 mentions) Special 96 well e plate, by Agilent Technologies (1 mentions) Mouse il 2, by Thermo Fisher Scientific (1 mentions) Ifn γ elisa kit, by Mabtech (1 mentions) Easysep mouse t cell isolation kit, by STEMCELL (1 mentions) Celltrace cfse cell proliferation kit, by Thermo Fisher Scientific (1 mentions) Facscanto 2, by BD (1 mentions)

Close spelling variants of this product

Mouse IL-15/IL-15Ra Recombinant mouse IL-7 and IL-15 Recombinant human IL-2 or mouse IL-15 Mouse IL-15/IL-15R Complex ELISA Ready-Set-Go Mouse IL-15/IL15R Platinum ELISA Recombinant mouse IL-15 IL-15

6 protocols using mouse il 15

Intestinal Organoid-Intraepithelial Lymphocyte Coculture

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We followed a modified protocol from a previous report (27 (link)). Briefly, intestinal organoids were cultured for 2 days prior to the co-culture with IELs. On the first day of co-culture, SI-IELs collected from the WT mice aged 9-weeks old or 89 weeks old were stained with anti-mouse CD3e-APC-Cy7, anti-mouse CD4-PE-Cy7, and the CD3⁺ (CD3⁺) and CD4⁺ (CD3⁺ CD4⁺) -IELs were sorted using FACSMelody (BD Bioscience). Cultured organoids released from Matrigel were washed and counted. We combined 200 organoids and 2.0 × 10⁵ IELs and centrifuged the samples for 3 min at 200 g. In the control group, the same number of organoids as the co-culture group was centrifuged. The pellet was suspended in 20 μL of Matrigel and placed in 24-well plates. After Matrigel polymerization, 500 μL of the complete medium with 100 U/mL recombinant human IL-2 (Roche), 10 ng/mL mouse IL-7 (Peprotech) and 10 ng/mL mouse IL-15 (Peprotech) were supplemented. The medium was refreshed every 1–2 days. Images of organoids were taken with a BZ-X710 microscope (Keyence).

Yonemoto Y., Nemoto Y., Morikawa R., Shibayama N., Oshima S., Nagaishi T., Mizutani T., Ito G., Fujii S, & Okamoto R. (2024). Single cell analysis revealed that two distinct, unique CD4+ T cell subsets were increased in the small intestinal intraepithelial lymphocytes of aged mice. Frontiers in Immunology, 15, 1340048.

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Optimized CTL Generation and Cytokine Recall

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Lymph node (LN) OT-I T cells were cultured in RPMI-1640 medium (Invitrogen) supplemented with 10% FCS, L-glutamine, antibiotics and 50 μM 2-mercaptoethanol. SIINFEKL (N4), SIITFEKL (T4) and SIIGFEKL (G4) peptides (Peptide Synthesis) were added to culture media at the concentrations stated in figure legends. In some experiments, cells were cultured in the presence of 100 nM rapamycin or 10 μM S6K1 inhibitor PF-4708671 (both Tocris Bioscience). These conditions have previously been optimized for the selective inhibition of target kinases by the drugs (28 (link)-30 (link)). For CTL generation, OT-I T cells were stimulated with 10 nM N4 for 2 d, washed, then differentiated in the presence of either recombinant human IL-2 or mouse IL-15 (both Peprotech) for a further 4d. For cytokine recall responses, in vitro-generated CTLs or ex vivo polyclonal splenic T cells from Lm-Ova infected mice were re-stimulated with peptide for 4 h in the presence of 2.5 μg/mL Brefeldin A (Sigma Aldrich).

Salmond R.J., Brownlie R.J., Meyuhas O, & Zamoyska R. (2015). Mechanistic Target of Rapamycin Complex 1/S6 Kinase 1 Signals Influence T Cell Activation Independently of Ribosomal Protein S6 Phosphorylation. The Journal of Immunology Author Choice, 195(10), 4615-4622.

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Assessing NK and Macrophage Cytotoxicity

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NK cell killing and macrophage killing assays were performed on the XCelligence SP platform and MP platform (ACEA Biosciences). Special 96-well E-plates (ACEA Biosciences) were coated with collagen (Sigma-Aldrich) and 4 × 10⁵ WT, B2m⁻^/⁻Ciita⁻^/⁻, or B2m⁻^/⁻Ciita⁻^/⁻ Cd47 tg miECs and plated in 100 µl cell-specific medium. After the cell index value reached 0.7, BALB/c, WT C57BL/6, or Sirpa⁻^/⁻ C57BL/6 NK cells or macrophages were added at an E:T ratio of 0.5:1, 0.8:1, or 1:1 with or without 1 ng/ml mouse IL-2 or 1 ng/ml mouse IL-15 (both PeproTech). As a negative control, cells were treated with 2% Triton X-100 in cell-specific media (data not shown). Some wells were treated with rabbit anti-mouse Pirb antibody (polyclonal, catalog no. MBS1489822; MyBioSource) at a concentration of 50 ng/ml. In some experiments, anti-mouse Cd47 blocking antibody (clone MIAP301, rat IgG2a,κ; BioXCell) and mouse FcR block (catalog no. 130–092-575, concentration 1:5; Miltenyi) or anti-Sirpα blocking antibody (clone P84, rat IgG1,κ; BioLegend) was used. Data were standardized and analyzed with RTCA software (ACEA Biosciences).

Deuse T., Hu X., Agbor-Enoh S., Jang M.K., Alawi M., Saygi C., Gravina A., Tediashvili G., Nguyen V.Q., Liu Y., Valantine H., Lanier L.L, & Schrepfer S. (2021). The SIRPα–CD47 immune checkpoint in NK cells. The Journal of Experimental Medicine, 218(3), e20200839.

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Generating Mouse Organoids and opT Cells

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Mouse organoids were generated from triple-negative breast cancer mouse models.21 (link) Autologous mouse T cells from the spleen were isolated by EasySep Mouse T Cell Isolation Kit (Stemcell Technologies, 19851). To generate opT, mouse organoids, and T cells were cultured in mouse T cell medium: Serum-free medium (CellGernix, 20 801–0100), 10% fetal bovine serum (FBS) (Glico, 16140071), mouse interleukin (IL)-2 (1000 IU/mL) (Peprotech, 400–02), mouse IL-15 (10 ng/mL, Peprotech, 400–24) and mouse IL-21 (10 ng/mL, Prospec, CYT-033). For killing assay, the organoids from mouse tumors or normal tissue were co-cultured with opT cells. The supernatants from the co-culture were harvested, and secreted IFN-γ was quantified by IFN-γ ELISA kit (Mabtech, 3421–1 H-6). Please see online supplemental methods for further details.

Meng Q., Xie S., Gray G.K., Dezfulian M.H., Gandarilla O., Li W., Huang L., Akshinthala D., Ferrer E., Conahan C., Perea Del Pino S., Grossman J., Elledge S.J., Hidalgo M, & Muthuswamy S.K. (2021). Empirical identification and validation of tumor-targeting T cell receptors from circulation using autologous pancreatic tumor organoids. Journal for Immunotherapy of Cancer, 9(11), e003213.

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Activation and Differentiation of OT-I CTLs

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Lymph node (LN) OT-I T cells were cultured in RPMI 1640 medium (Invitrogen) supplemented with 10% FCS, l-glutamine, antibiotics, and 50 μM 2-ME. SIINFEKL (N4), SIITFEKL (T4) and SIIGFEKL (G4) peptides (Peptide Synthesis) were added to culture media at the concentrations stated in figure legends. In some experiments, cells were cultured in the presence of 100 nM rapamycin or 10 μM S6K1 inhibitor PF-4708671 (both Tocris Bioscience). These conditions have previously been optimized for the selective inhibition of target kinases by the drugs (28 (link)–30 (link)). For CTL generation, OT-I T cells were stimulated with 10 nM N4 for 2 d, washed, and then differentiated in the presence of either recombinant human IL-2 or mouse IL-15 (both PeproTech) for an additional 4d. For cytokine recall responses, in vitro–generated CTLs or ex vivo polyclonal splenic T cells from Lm-Ova–infected mice were restimulated with peptide for 4 h in the presence of 2.5 μg/ml brefeldin A (Sigma-Aldrich).

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Proliferation Assay of IEL and SP CD4+ Cells

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1 x 10⁵ of sort-purified IEL- or SP CD4⁺ cells were labelled with 2.5μM CFSE (CellTrace CFSE Cell Proliferation Kit, ThermoFisher SCIENTIFIC), and then stimulated with PMA+ionomycin PMA+ionomycin (PMA 50ng/ml, ionomycin 500ng/ml) in the complete medium with 100 U/mL recombinant human IL2 (Roche), 50 ng/mL mouse IL7 (Peprotech), and 50 ng/mL mouse IL15 (Peprotech) under the condition of 37°C, 5% CO2. After 72 hours, cells were harvested and stained with appropriate antibodies and analyzed by FACS Canto II (BD Bioscience).

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