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Recombinant human il 2 proleukin

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Recombinant human IL-2 (Proleukin) is a laboratory product that contains the human cytokine interleukin-2. Interleukin-2 is a protein involved in the regulation of immune responses.

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Lab products found in correlation

Rpmi 1640, by Thermo Fisher Scientific (4 mentions) Sodium pyruvate, by Thermo Fisher Scientific (3 mentions) Hepes, by Thermo Fisher Scientific (3 mentions) Mouse pan t cell isolation kit, by Miltenyi Biotec (3 mentions) Mem non essential amino acids 1, by Thermo Fisher Scientific (3 mentions) Mouse anti cd3 28 dynabeads, by Thermo Fisher Scientific (3 mentions) L glutamine, by Thermo Fisher Scientific (3 mentions) Phytohemagglutinin (pha), by Merck Group (2 mentions) Easysep mouse streptavidin rapidspheres isolation kit, by STEMCELL (1 mentions) Glutamine, by Thermo Fisher Scientific (1 mentions) Recombinant mouse il 12, by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) β mercaptoethanol, by Merck Group (1 mentions) Anti cd4 apc, by BD (1 mentions) Anti cd44 pe, by BD (1 mentions) Anti cd45 apc, by BD (1 mentions) Anti b220 fitc, by BD (1 mentions) Anti cd8 pe cy5, by BD (1 mentions) Celltrace cfse cell proliferation kit, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Proleukin (89 citations) Recombinant human IL-2 (44 citations) IL-2 (Proleukin; (25 citations) Human IL-2 (12 citations) Recombinant IL-2 (5 citations)

9 protocols using recombinant human il 2 proleukin

1

Activation and Culture of Mouse CD4+ T Cells

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T cells were activated and cultured at 37°C with 5% CO2 in RPMI 1640 containing glutamine (Invitrogen), supplemented with 10% FBS (Gibco), penicillin/streptomycin (Gibco), and 50 μM β‐mercaptoethanol (Sigma) unless otherwise indicated. Spleen was homogenized, red blood cells lysed, and CD4+ T cells depleted using the EasySep Mouse Streptavidin RapidSpheres Isolation Kit from StemCell Technologies. Cells were then cultured at a cell density of 2 million/ml in the presence of CD3 (2C11; 1 μg/ml) and CD28 (37.51; 0.5 μg/ml) antibodies supplemented with 20 ng/ml recombinant human IL‐2 (Proleukin, Novartis) and 2 ng/ml recombinant mouse IL‐12 (Peprotech) for 48 h. Then, cells were washed and further cultured in media with 20 ng/ml IL‐2, regularly splitting them to 0.3 × 106/ml.
Ventura P.M., Gakovic M., Fischer B.A., Spinelli L., Rota G., Pathak S., Khameneh H.J., Zenobi A., Thomson S., Birchmeier W., Cantrell D.A, & Guarda G. (2022). Concomitant deletion of Ptpn6 and Ptpn11 in T cells fails to improve anticancer responses. EMBO Reports, 23(11), e55399.
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2

Mouse and Human gp100 Peptide Protocol

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Mouse gp10025–33 (mgp100, EGSRNQDWL) and human gp10025–33 (hgp100, KVPRNQDWL) peptides were synthesized by Peptron (Daejeon, Korea). CD8 microbeads were purchased from Miltenyi Biotec (Auburn, AL, USA). All antibodies used for flow cytometry were purchased from BD Bioscience, including anti-Thy1.1-FITC, anti-CD3-FITC, anti-B220-FITC, anti-CD62L-FITC, anti-Ly-6C-FITC, anti-CD19-PE, anti-CD8-PE, anti-CD44-PE, anti-CD8-PE-Cy5, anti-Thy1.1-PE-Cy5, anti-CD11b-PE-Cy5, anti-CD4-APC, anti-CD8-APC, and anti-CD45-APC. Recombinant human IL-2 (Proleukin) was purchased from Novartis, and the Fc fusion protein of human IL-7 (IL7-Fc), which consist of the extracellular domain of human IL-7 (aa 26–177) fused to the N-terminus of the Fc portion of a mutant human IgG1, was obtained from AdipoGen (Seoul, Korea). The CellTrace CFSE Cell Proliferation Kit was purchased from Invitrogen (Carlsbad, CA, USA).
Yu E.M., Cho E., Singh R., Kim S.H., Han C., Han S., Lee D.G., Kim Y.H., Kwon B.S, & Choi B.K. (2021). IL7-Fc Enhances the Efficacy of Adoptive T Cell Therapy under Lymphopenic Conditions in a Murine Melanoma Model. Cells, 10(8), 2018.
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3

Isolation and Activation of NK Cells

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As previously described, fresh NK cells were isolated from peripheral blood mononuclear cells (PBMC) obtained from buffy coats [28 (link)]. Buffy coats were collected from volunteer blood donors admitted to the blood transfusion service of OPBG after obtaining informed consent. The Ethical Committee of OPBG approved the study (ID AIRC5x1000 #21147) that was conducted in accordance with the ethical principles stated in the Declaration of Helsinki.
To obtain polyclonal activated NK (aNK) cells, freshly isolated NK cells were cultured on 30 Gy irradiated allogeneic PBMC feeder cells in the presence of 600 U/mL recombinant human IL-2 (Proleukin; Novartis-Farma, Basel, Switzerland) and 1.5 ng/mL phytohemagglutinin (Merck-Millipore, Burlington, MA, USA) for the first week, as previously described [28 (link)]. The aNKs were used to perform experiments during the exponential growth phase.
Di Matteo S., Avanzini M.A., Pelizzo G., Calcaterra V., Croce S., Spaggiari G.M., Theuer C., Zuccotti G., Moretta L., Pelosi A, & Azzarone B. (2022). Neuroblastoma Tumor-Associated Mesenchymal Stromal Cells Regulate the Cytolytic Functions of NK Cells. Cancers, 15(1), 19.
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4

Expansion of Prostate Tumor-Infiltrating Lymphocytes

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Prostate punches and TRUS-Bx biopsies were cut into multiple fragments of about 0.5–1 mm3 of size. Each fragment was placed into one well of a 24-well plate with 2 ml complete medium (CM) comprised of RPMI 1640 medium (Gibco, Thermo Fisher Scientific, Waltham, MA) containing 10% heat-inactivated human serum (Valley Biomedical, Winchester, VA or Gemini Bio, West Sacramento, CA), 25 mmol/l HEPES pH 7.2 (Gibco), 50 μg/ml gentamycin (Gentamicin IKA, Teva, Israel), 100 U/ml penicillin and 100 mg/ml streptomycin (Gibco) and supplemented with 3,000 IU/ml recombinant human IL-2 (Proleukin, Novartis Pharma, Germany). Prostate-TIL were allowed to extravasate from the fragments. Half of the media was replaced every 2 to 3 days. Following the establishment of TIL cultures, a standard 14-days Rapid Expansion Procedure (REP) was initiated by stimulating prostate-TIL with 30 ng/ml anti-CD3 antibody MACS GMP CD3 pure (clone OKT-3; Miltenyi Biotech, Germany), 3,000 IU/ml IL-2, and irradiated peripheral blood mononuclear cells from non-related donors as feeder cells (5000 rad, 100:1 ratio between feeder cells and TIL) in 50% CM, 50% AIM-V medium (Invitrogen, Thermo Fisher Scientific, Waltham, MA) (for details see Besser MJ28; Itzhaki O29). REP was performed in T25 flasks or 24-well plates.
Yunger S., Bar El A., Zeltzer L.A., Fridman E., Raviv G., Laufer M., Schachter J., Markel G., Itzhaki O, & Besser M.J. (2019). Tumor-infiltrating lymphocytes from human prostate tumors reveal anti-tumor reactivity and potential for adoptive cell therapy. Oncoimmunology, 8(12), e1672494.
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5

UCB-Derived T Cell Expansion Protocol

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Fresh UCB units not meeting cell dose criteria for public banking were obtained from the Cleveland Cord Blood Center (Cleveland, OH, USA), and mononuclear cells were separated using density gradient centrifugation with Accu-prep (axis-Shield PoC AS, Oslo, Norway). T cells were isolated, activated and expanded with Dynabeads ClinExVivo CD3/CD28 magnetic beads (Invitrogen), according to the manufacturer's instructions. UCB-derived T cells were cultured in RPMI 1640 supplemented with 10% fetal calf serum, 100 U/ml penicillin and streptomycin, and 2 mm L-glutamine, in the presence of 100 IU/ml recombinant human IL-2 (Proleukin, Novartis, Basel, Switzerland), 10 ng/ml recombinant human IL-12, 10 ng/ml recombinant human IL-7 or 10 ng/ml recombinant human IL-15 (all from R&D Systems, Minneapolis, MN, USA). Viable cells were enumerated using Trypan blue (Invitrogen) exclusion. Secondary stimulation was achieved with ClinExVivo CD3/CD28 beads at a bead to T-cell ratio of 1:2.
Pegram H., Purdon T., van Leeuwen D., Curran K., Giralt S., Barker J, & Brentjens R. (2014). IL-12-secreting CD19-targeted cord blood-derived T cells for the immunotherapy of B-cell acute lymphoblastic leukemia. Leukemia, 29(2), 415-422.
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6

Isolation and Activation of Mouse T Cells

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B6.SJL-Ptrca/BoyAiTac mice (CD45.1 mice) were euthanized and spleens were collected. After tissue dissection and red blood cell lysis, primary mouse T cells were purified using the mouse Pan T cell Isolation Kit (Miltenyi Biotec). Purified T cells were cultured in RPMI-1640 (Invitrogen) supplemented with 10% FBS (HyClone), 10 mM HEPES (Invitrogen), 2 mM l-glutamine (Invitrogen), MEM non-essential amino acids 1× (Invitrogen), 55 μM β-mercaptoethanol, 1 mM sodium pyruvate (Invitrogen), 100 IU ml−1 recombinant human IL-2 (Proleukin; Novartis) and mouse anti-CD3/28 Dynabeads (Gibco) at a bead:cell ratio of 1:2. T cells were spinoculated with retroviral supernatant collected from Phoenix-ECO cells 24 h after initial T cell activation as described44 (link),45 (link) and used for functional analysis 3–4 days later.
Amor C., Fernández-Maestre I., Chowdhury S., Ho Y.J., Nadella S., Graham C., Carrasco S.E., Nnuji-John E., Feucht J., Hinterleitner C., Barthet V.J., Boyer J.A., Mezzadra R., Wereski M.G., Tuveson D.A., Levine R.L., Jones L.W., Sadelain M, & Lowe S.W. (2023). Prophylactic and long-lasting efficacy of senolytic CAR T cells against age-related metabolic dysfunction. Research Square.
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7

Isolation and Expansion of Mouse T Cells

Cited 1 time
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B6.SJL-Ptrca/BoyAiTac mice (CD45.1 mice) were euthanized and spleens were harvested. Following tissue dissection and red blood lysis, primary mouse T cells were purified using the mouse Pan T cell Isolation Kit (Miltenyi Biotec). Purified T cells were cultured in RPMI-1640 (Invitrogen) supplemented with 10% fetal bovine serum (FBS; HyClone), 10 mM HEPES (Invitrogen), 2 mM L-glutamine (Invitrogen), MEM nonessential amino acids 1× (Invitrogen), 55 uM β-mercaptoethanol, 1 mM sodium pyruvate (Invitrogen), 100 IU/ mL of recombinant human IL-2 (Proleukin; Novartis) and mouse anti-CD3/28 Dynabeads (Gibco) at a bead:cell ratio of 1:2. T cells were spinoculated with retroviral supernatant collected from Phoenix-ECO cells 24 hours after initial T cell expansion as described33 (link),53 (link) and used for functional analysis 3–4 days later.
Amor C., Feucht J., Leibold J., Ho Y.J., Zhu C., Alonso-Curbelo D., Mansilla-Soto J., Boyer J.A., Li X., Giavridis T., Kulick A., Houlihan S., Peerschke E., Friedman S.L., Ponomarev V., Piersigilli A., Sadelain M, & Lowe S.W. (2020). Senolytic CAR T cells reverse senescence-associated pathologies. Nature, 583(7814), 127-132.
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8

Isolation and Expansion of Mouse T Cells

Cited 1 time
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B6.SJL-Ptrca/BoyAiTac mice (CD45.1 mice) were euthanized and spleens were harvested. Following tissue dissection and red blood lysis, primary mouse T cells were purified using the mouse Pan T cell Isolation Kit (Miltenyi Biotec). Purified T cells were cultured in RPMI-1640 (Invitrogen) supplemented with 10% fetal bovine serum (FBS; HyClone), 10 mM HEPES (Invitrogen), 2 mM L-glutamine (Invitrogen), MEM nonessential amino acids 1× (Invitrogen), 55 uM β-mercaptoethanol, 1 mM sodium pyruvate (Invitrogen), 100 IU/ mL of recombinant human IL-2 (Proleukin; Novartis) and mouse anti-CD3/28 Dynabeads (Gibco) at a bead:cell ratio of 1:2. T cells were spinoculated with retroviral supernatant collected from Phoenix-ECO cells 24 hours after initial T cell expansion as described33 (link),53 (link) and used for functional analysis 3–4 days later.
Amor C., Feucht J., Leibold J., Ho Y.J., Zhu C., Alonso-Curbelo D., Mansilla-Soto J., Boyer J.A., Li X., Giavridis T., Kulick A., Houlihan S., Peerschke E., Friedman S.L., Ponomarev V., Piersigilli A., Sadelain M, & Lowe S.W. (2020). Senolytic CAR T cells reverse senescence-associated pathologies. Nature, 583(7814), 127-132.
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9

Isolation and Activation of Polyclonal NK Cells

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Fresh NK cells were isolated from peripheral blood mononuclear cells (PBMC) obtained from buffy coats as previously described.24 (link) To obtain polyclonal activated NK (aNK) cells, freshly isolated NK cells were cultured on 30 Gy irradiated allogeneic PBMC feeder cells in the presence of 600 U/mL recombinant human IL-2 (Proleukin; Novartis-Farma, Origgio, Italy) and 1.5 ng/mL phytohemagglutinin (Merck-Millipore, Burlington, USA) for the first week, as previously described.24 (link) aNKs were used to perform experiments during the exponential growth phase.
Canzonetta C., Pelosi A., Di Matteo S., Veneziani I., Tumino N., Vacca P., Munari E., Pezzullo M., Theuer C., De Vito R., Pistoia V., Tomao L., Locatelli F., Moretta L., Caruana I, & Azzarone B. (2021). Identification of neuroblastoma cell lines with uncommon TAZ+/mesenchymal stromal cell phenotype with strong suppressive activity on natural killer cells. Journal for Immunotherapy of Cancer, 9(1), e001313.
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