Il 15

Manufactured by ProSpec

Sourced in Israel

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

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Lab products found in correlation

Interleukin 2 (il 2), by ProSpec (3 mentions) Il 21, by ProSpec (3 mentions) Medimachine, by BD (1 mentions) Recombinant il 2, by ProSpec (1 mentions) Penicillin streptomycin, by Merck Group (1 mentions) Interleukin 3 (il 3), by R&D Systems (1 mentions) Ascorbic acid, by Merck Group (1 mentions) Flt 3 ligand, by ProSpec (1 mentions) Interleukin 7 (il 7), by ProSpec (1 mentions) Heat inactivated human ab serum, by Merck Group (1 mentions) Il 15, by R&D Systems (1 mentions) Ham s f 12 medium, by Lonza (1 mentions) Interleukin 2 (il 2), by BD (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Amphotericin b, by Merck Group (1 mentions) Anti cd3 okt3, by BioLegend (1 mentions) Human ab serum, by Innovative Research (1 mentions) Serum free medium, by CellGenix (1 mentions) Vmax kinetic microplate reader, by Molecular Devices (1 mentions)

7 protocols using il 15

NK Cell Proliferation Assay

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NK cells were labeled with 2 μmol/L carboxyfluorescein succinimidyl ester (CFSE) per 1 × 10 6 cells (Invitrogen). CBCD34 + and PBNK cells were then stimulated with 200 IU IL-2 (Prospec) and CBNK cells were stimulated with 1000 IU IL-2.
Comparatively cells were also stimulated with 20 ng/mL IL-15 (Prospec) or primed with CTV-1 lysate. Proliferation was assessed on days 2, 5 and 7 post stimulation by flow cytometry.

Domogala A., Blundell M., Thrasher A., Lowdell M.W., Madrigal J.A, & Saudemont A. (2017). Natural killer cells differentiated in vitro from cord blood CD34⁺ cells are more advantageous for use as an immunotherapy than peripheral blood and cord blood natural killer cells. Cytotherapy, 19(6).

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Expansion of Tumor-Infiltrating Lymphocytes from Glioma Tissue

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Plasma was removed following centrifugation of whole blood samples, and stored at -80°C. Glioma tumor tissue was harvested in the course of tumor surgery at the Department of Neuro-Oncology at Karolinska University Hospital, Stockholm. Tumor tissue was immediately transferred to Cellgro GMP Serum-free DC medium medium supplemented with 5% pooled human AB serum. Tumor tissue was dissected into fragments of approximately 1-2mm3 using a sterile scalpel, or processed into a tissue homogenate using the BD Medimachine. Tissue fragments of the cell suspension were washed twice with PBS and cultured in 24 well plates in Cellgro medium plus 5% pooled human AB serum supplemented with recombinant IL-2 (1000IU/ml), IL-15(10ng/ml) and IL-21 (10ng/ml) (Prospec, Ness-Ziona, Israel). Medium was changed as necessary. Irradiated (55Gy) feeder cells (allogeneic PBMCs) at the ratio of 1 (feeder cells):10 (TILs) was added on day 7. OKT3 (anti-CD3 monoclonal antibody, BioLegend, San Diego, CA) was used at 10ng/ml as TILs became visible under the microscope. TILs were transferred to 6-well plates; upon achieving > 70% confluence in the 24-well surface. They were further expanded in G-Rex flasks (Wilson Wolf, New Brighton, MN) using 30ng OKT3/mL and irradiated (55Gry) allogeneic feeder cells at the ratio of 1 (feeder cells):10 (TILs).

Liu Z., Rao M., Poiret T., Nava S., Meng Q., von Landenberg A., Bartek J J.r., Xie S., Sinclair G., Peredo I., Dodoo E, & Maeurer M. (2017). Mesothelin as a novel biomarker and immunotherapeutic target in human glioblastoma. Oncotarget, 8(46), 80208-80222.

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Hematopoietic Stem Cell Expansion

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EL08.1D2 cells were cultured as previously described [28] (link). HSC were plated over 2000 irradiated EL08.1D2 cells in a medium consisting of 2∶1 (vol:vol) mix of Dulbecco’s Modification of Eagle’s Medium (DMEM) with 4.5 g/L glucose, L-glutamine 1640, and sodium pyruvate/Ham’s F12 Medium (all from Lonza, Verviers, Belgium) supplemented with 50 µM beta mercaptoethanol (β-ME), 50 µM ethanolamine, 20 mg/L ascorbic acid, 50 µg/L sodium selenite, 1% penicillin/streptomycin and 20% heat-inactivated human AB serum (all from Sigma, Poole, UK). Different interleukins such as 10 ng/ml IL-15, 5 ng/ml IL-3 (only for the first week; R&D System, Abingdon, UK), 20 ng/ml IL-7, 20 ng/ml c-kit ligand stem cell factor (SCF) and 10 ng/ml Flt3 ligand were added up to 21 days of culture and only 10 ng/ml IL-15 were added from day 21 to day 35 (all from Prospec, Israel). Cultures underwent weekly hemi-depletion. Lymphoid progenitors were characterized as CD45+CD7+ whereas myeloid progenitors as CD45+CD33+.

Lee F., Luevano M., Veys P., Yong K., Madrigal A., Shaw B.E, & Saudemont A. (2014). The Effects of CAMPATH-1H on Cell Viability Do Not Correlate to the CD52 Density on the Cell Surface. PLoS ONE, 9(7), e103254.

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Modeling Esophageal Inflammation in Organotypic Culture

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Organotypic culture was performed as previously described [48 (link), 53 (link)]. Human esophageal keratinocytes were seeded on a collagen fibroblast layer with and without human peripheral blood mononuclear cells (PBMC), which were stimulated by inflammatory cytokines (IL-2, IL-7, and IL-15; Human Immunology Core, University of Pennsylvania) as described [54 (link)]. To model a T_H1 inflammatory environment, the pro-inflammatory cytokines IL-7 (10 ng/mL; Cell Signaling) and IL-15 (20 ng/mL; Prospec-Tany Technogene) were included in cell culture media, and IL-2 (10 U/mL, BD Biosciences) was added to support PBMC viability. Organotypic cultures were processed and stained according to standard protocols [48 (link)].

Shaverdashvili K., Padlo J., Weinblatt D., Jia Y., Jiang W., Rao D., Laczkó D., Whelan K.A., Lynch J.P., Muir A.B, & Katz J.P. (2019). KLF4 activates NFκB signaling and esophageal epithelial inflammation via the Rho-related GTP-binding protein RHOF. PLoS ONE, 14(4), e0215746.

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Generating CMV-Specific T Cell Lines and Clones

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CMV-specific T cell line (HD-2) and T cell clones (A1-8 and A1-9) specific for CMV pp65 protein were generated from a single HLA-A*0201+ healthy control. CMV-specific T cells were stained with HLA-PE-conjugated A*0201 CMV_NLVPMVATV Dextramer (Immunex, Copenhagen, Denmark) for 30 min at room temperature. HLA-A*0201-CMV-CTL were then sorted using a FACS Aria flow cytometer.
Sorted CMV-CTL from HD-2 cell line were then clonally selected using limiting dilution in T cell medium containing Serum-free medium (Cell-Genix, Freiburg, Germany), with 10% human AB serum (Innovative Research, MI) supplemented with IL-2 (1000 IU/ml), IL-15 (10ng/ml), IL-21 (10ng/ml), (Prospec, Ness-Ziona, Israel), penicillin (100IU/ml), streptomycin (100μg/ml) (Life Technologies, Carlsbad, CA), and amphotericin B (2.5mg/L) (Sigma-Aldrich, St Louis, MO) and seeded with anti-CD3 (OKT3) (Biolegend, CA, USA) and addition of γ-irradiated allogeneic feeder cells at a 5:1 E:T ratio. After 14 days, the clonal selection was repeated. Expanded T cell clones were transferred into larger vessels when required where they were subsequently expanded in T cell culture medium and maintained by biweekly stimulation with OKT3 and irradiated allogenic feeder cells.

Luo X.H., Poiret T., Liu Z., Meng Q., Nagchowdhury A, & Ljungman P. (2023). Different recovery patterns of CMV-specific and WT1-specific T cells in patients with acute myeloid leukemia undergoing allogeneic hematopoietic cell transplantation: Impact of CMV infection and leukemia relapse. Frontiers in Immunology, 13, 1027593.

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Quantifying T-cell Activation by ELISA

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Whole blood was first diluted at a ratio of 1:1.5 with either RPMI 1640 medium containing l-glutamine (2 mM) with antibiotics (100 IU/ml penicillin and 100 µg/ml streptomycin) (Life Technologies, Carlsbad, USA) alone or supplemented with the following cytokine cocktail: IL-2 (1000 IU/ml), IL-15 (10 ng/ml) and IL-21 (10 ng/ml) (Prospec, Ness-Ziona, Israel). Diluted blood was added in duplicates for each condition to 96-well plates pre-coated with the proteins EBNA-1 or CMV-pp65 (Prospec, Ness-Ziona, Israel) at a final concentration of 1 μg/ml and incubated for 7 days at 37 °C with 5% CO2 as previously described [26 (link), 27 (link)]. Antigen-free medium was used as negative control while phytohemagglutinin protein (PHA 5 μg/ml, Sigma Aldrich) was used positive control. Cell culture supernatants were harvested after the incubation period to quantify IFNγ production by sandwich ELISA (Mabtech, Stockholm, Sweden) according to the manufacturer’s instructions. Assay plates were analyzed using a Vmax kinetic microplate reader (Molecular Devices, USA) at 450 nm. After basal IFNγ production (medium only) subtraction, data were reported as absolute cytokine concentration values (pg/ml).

Liu Z., Poiret T., Meng Q., Rao M., von Landenberg A., Schoutrop E., Valentini D., Dodoo E., Peredo-Harvey I, & Maeurer M. (2018). Epstein–Barr virus- and cytomegalovirus-specific immune response in patients with brain cancer. Journal of Translational Medicine, 16, 182.

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Isolation and Activation of CD8+ T Cells

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Four healthy males (three in their twenties and one in his fifties) and one female (in her twenties) donated 10 cc of blood in compliance with the protocol approved by the IRB of Sookmyung Women’s University (SM-IRB-08-0225). CD8⁺ T cells were purified using the Dynal CD8⁺ Isolation Kit (Invitrogen, USA) and activated by the treatment of Dynabeads conjugated with anti-CD3 and anti-CD28 antibodies (Invitrogen). The Roswell Park Memorial Institute (RPMI) medium was replaced every two days with fresh supplements of human IL-2 (60 UI/ml; Sigma-Aldrich, USA) and IL-15 (5 ng/ml; ProSpec, USA). At the start of the activation, 5 mM NAM was added. The individual donors provided written informed consent to publish these case details.

Choi H.J., Jang S.Y, & Hwang E.S. (2015). High-Dose Nicotinamide Suppresses ROS Generation and Augments Population Expansion during CD8+ T Cell Activation. Molecules and Cells, 38(10), 918-924.

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