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Recombinant human il 15

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Recombinant human IL-15 is a cytokine that is produced and purified from E. coli. It is a member of the common gamma-chain cytokine family and is involved in the proliferation and activation of natural killer cells and memory T cells.

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Lab products found in correlation

Recombinant human il 2, by Novartis (3 mentions) Celltiter 96 aqueous one solution cell proliferation assay, by Promega (1 mentions) Glutamax 1, by Thermo Fisher Scientific (1 mentions) Aim 5, by Thermo Fisher Scientific (1 mentions) Macs nk cell isolation kit, by Miltenyi Biotec (1 mentions) Cd3 cd28 dynabead, by Thermo Fisher Scientific (1 mentions) X vivo 15, by Lonza (1 mentions) X vivo 15 media, by Lonza (1 mentions) Dynabeads human t expander cd3 cd28, by Thermo Fisher Scientific (1 mentions)

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IL-15 (15 citations)

5 protocols using recombinant human il 15

1

Isolation and Expansion of Central Memory T Cells

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Isolation and culture of human PBMC has been previously described.14 15 (link) Central memory T (Tcm) cells were enriched by negative selection of CD14, CD25, and CD45RA and positive selection of CD62L. For expansion, cells were maintained in complete X-VIVO media with 50 U/mL recombinant human IL2 (Novartis) and 0.5 ng/mL recombinant human IL15 (CellGenix).
Kuo Y.C., Kuo C.F., Jenkins K., Hung A.F., Chang W.C., Park M., Aguilar B., Starr R., Hibbard J., Brown C, & Williams J.C. (2022). Antibody-based redirection of universal Fabrack-CAR T cells selectively kill antigen bearing tumor cells. Journal for Immunotherapy of Cancer, 10(6), e003752.
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2

Cytokine-Mediated NK Cell Proliferation

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1 × 105 freshly isolated bulk NK cells were cultured either alone, or with 200 IU/mL recombinant human IL2 (Chiron), 10 ng/mL recombinant human IL15 (CellGenix), or moDCpoly for 6 d. TG101348, ruxolitinib, or DMSO control was added once on day 0. NK-cell proliferation was determined by a colorimetric assay according to the manufacturer's instructions (CellTiter 96 AQueous One Solution Cell Proliferation Assay; Promega). Metabolically active cells reduce a tetrazolium compound, which is added for the final 4 h of the coculture, into a colored formazan product that is detected at 490nm.
Curran S.A., Shyer J.A., St. Angelo E.T., Talbot L.R., Sharma S., Chung D.J., Heller G., Hsu K.C., Betts B.C, & Young J.W. (2016). Human Dendritic Cells Mitigate NK-Cell Dysfunction Mediated by Nonselective JAK1/2 Blockade. Cancer immunology research, 5(1), 52-60.
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3

Isolation and Culture of NK Cells

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Peripheral blood mononuclear cells (PBMCs) were obtained from healthy blood-bank donors by Ficoll density gradient separation. NK cells were isolated using the MACS NK Cell Isolation Kit according to the manufacturer’s protocol (Miltenyi Biotech, Bergisch Gladbach, Germany). NK cell purity was 94–99 %. Freshly isolated, resting NK cells were cultured for 2–5 weeks in AIM-V supplemented with glutamax I (Life Technologies), 10 % pooled human serum (Sanquin, Rotterdam, the Netherlands), penicillin/streptomycin (100 U/mL and 100 μg/mL, respectively) and 10 ng/mL recombinant human IL-15 (Cellgenix, Freiburg, Germany).
Boerman G.H., van Ostaijen-ten Dam M.M., Kraal K.C., Santos S.J., Ball L.M., Lankester A.C., Schilham M.W., Egeler R.M, & van Tol M.J. (2015). Role of NKG2D, DNAM-1 and natural cytotoxicity receptors in cytotoxicity toward rhabdomyosarcoma cell lines mediated by resting and IL-15-activated human natural killer cells. Cancer Immunology, Immunotherapy, 64(5), 573-583.
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4

Activation and Transduction of T Cells for CAR Therapy

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T cell activation and transduction was performed as described previously.35 (link) Briefly, freshly thawed CD14– or whole PBMCs were washed once and cultured in X-VIVO-15 (Lonza) with 10% FBS (complete X-VIVO) containing 100 U/mL recombinant human IL-2 (Novartis Oncology) and 0.5 ng/mL recombinant human IL-15 (CellGenix). For CAR lentiviral transduction, T cells were cultured with CD3/CD28 Dynabeads (Life Technologies), protamine sulfate (APP Pharmaceuticals), cytokine mixture (as stated above), and desired lentivirus at a 0.1–1 multiplicity of infection the day following stimulation. Cells were then cultured in and replenished with fresh complete X-VIVO containing cytokines every 2–3 days. After 7 days, beads were magnetically removed, and cells were further expanded in complete X-VIVO containing cytokines to achieve desired cell yield. CAR T cells were positively selected for truncated CD19 using the EasySep CD19 Positive Enrichment Kit I or II (StemCell Technologies) (for PSCA-CAR T cells) or positively selected for truncated EGFR using a custom EasySep EGFR Positive Enrichment Kit (for CD19-CAR T cells) according to the manufacturer’s protocol. Following further expansion, cells were frozen in CryoStor CS5 prior to in vitro and in vivo studies. Purity and phenotype of CAR T cells were verified by flow cytometry.
Yamaguchi Y., Gibson J., Ou K., Lopez L.S., Ng R.H., Leggett N., Jonsson V.D., Zarif J.C., Lee P.P., Wang X., Martinez C., Dorff T.B., Forman S.J, & Priceman S.J. (2022). PD-L1 blockade restores CAR T cell activity through IFN-γ-regulation of CD163+ M2 macrophages. Journal for Immunotherapy of Cancer, 10(6), e004400.
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5

Engineering Targeted T-Cell Therapies

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Naïve and memory T cells were isolated from healthy donors at City of Hope under protocols approved by the COH IRB (6 (link),42 ). The construct of IL13Rα2-targeted CAR and CAR transduction was described in previous studies (6 (link),20 (link)), (7 ). In brief, primary T cells were stimulated with Dynabeads Human T expander CD3/CD28 (Invitrogen) (T cells: beads = 1:3) for 24 hours and transduced with CAR lentivirus (multiplicity of infection [MOI] = 0.3). 7 days after CAR transduction, CD3/CD28 beads were removed and cells were resuspended and expanded in X-VIVO 15 media (Lonza) containing 10% FCS, 50 U/ml recombinant human IL-2 (Novartis), and 0.5 ng/ml recombinant human IL-15 (CellGenix) for additional 10-15 days before cryopreservation.
Alizadeh D., Wong R.A., Gholamin S., Maker M., Aftabizadeh M., Yang X., Pecoraro J.R., Jeppson J.D., Wang D., Aguilar B., Starr R., Larmonier C.B., Larmonier N., Chen M.H., Wu X., Ribas A., Badie B., Forman S.J, & Brown C.E. (2021). IFNγ is Critical for CAR T Cell Mediated Myeloid Activation and Induction of Endogenous Immunity. Cancer discovery, 11(9), 2248-2265.
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