Recombinant human IL-15 protein - Bioactive cytokine

Human IL-15 protein - Mammalian cell-expressed, tag-free, carrier-free

Recombinant human IL-15 is a high-quality and biologically active cytokine, validated using proprietary IL-15 reporter cells. This immunomodulating cytokine of the γc superfamily is produced in CHO cells to ensure protein glycosylation and bona fide 3D structure.

Recombinant human IL-15 can be used together with HEK-Blue™ IL-2/IL-15 cells, a cell line that detects human IL-15, for the screening of inhibitory molecules, such as Ordesekimab, a therapeutic monoclonal antibody targeting the IL-15 cytokine (see figures).

IL-15 signaling and biological functions

Key features

• Each lot is validated using HEK-Blue™ IL-2/IL-15 cells
• Endotoxin < 0.01 EU/µg
• 0.2 µm sterile-filtered

Applications

• T cell and NK cell activation and propagation
• IL-15 detection and quantification assays (positive control)
• Screening and release assays for antibodies blocking IL-15 signaling
• Screening and release assays for engineered IL-15 proteins

Interleukin 15 (IL-15) is an immunomodulating cytokine that is structurally and functionally related to IL-2. IL-15 promotes the ontogeny, activation, and survival of NK cells, NKT cells, T cells, and B cells. IL-15 also plays controversial roles in the development and progression of various hematological malignancies. Recombinant IL-15 can be used in combination with other cytokines like IL-2 or IL-7 to increase the efficiency of T and NK cell expansion and maturation in cell culture protocols.

 More details

InvivoGen’s products are for research use only, and not for clinical or veterinary use.

Figures

SDS-PAGE analysis of recombinant human (h)IL-15. 3 μg of hIL-15 was loaded on a 12% Mini-PROTEAN® TGX Stain-Free™ Precast Gel (Bio-Rad). Detection was performed as per the manufacturer’s instructions. Since IL-15 is highly glycosylated and the production of recombinant proteins in mammalian cells ensures protein glycosylation, the SDS gel shows a characteristic smear.

Dose-response of HEK-Blue™ IL-2/IL-15 cells to recombinant human IL-15. HEK-Blue™ IL-2/IL-15 cells were stimulated with increasing concentrations of recombinant human IL-15. After overnight incubation, the STAT5-induced SEAP activity was determined using QUANTI-Blue™, a SEAP detection reagent. Data are shown as optical density (OD) at 650 nm (mean ± SEM).

Dose-dependent inhibition of HEK-Blue™ IL-2/IL-15 cell response using Ordesekimab biosimilar. Increasing concentrations of Anti-hIL-15-hIgG1 (0.1 ng/ml - 10 µg/ml) were incubated with recombinant human (h)IL-15 (30 pg/ml) for 1 h before the addition of HEK-Blue™ IL-2/IL-15 cells. After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown in percentage of activity (%) (mean ± SEM).

Specifications

Source: Chinese hamster ovary (CHO) cells

Species: Human

Alternative name: IL15, Interleukin-15 

Carrier: Carrier-free

Tag: Tag-free

Accession number: P40933

Protein size: 114 a.a. (N49-S162)

Molecular weight: ~ 13 kDa (SDS-PAGE)

Solubility: 100 μg/ml in water

Formulation: Phosphate buffer saline (pH 7.4), 5% saccharose

Sterility: 0.2 µm filtration

Form:  Lyophilized

Reconstitution buffer:  Endotoxin-free water (provided)

Purity:  ≥95% (SDS-PAGE)

Endotoxin level:  ≤ 0.01 EU/μg (measurement by kinetic chromogenic LAL assay)

Tested applications: Cellular assays

Quality control: Each lot is functionally tested and validated

Contents

Recombinant human IL-15 is provided lyophilized and is available in two quantities:

• rcyc-hil15: 10 µg
• rcyc-hil15-5: 50 µg (5 x 10 µg)

Each product is provided with 1.5 ml endotoxin-free water.

 Recombinant human IL-15 is shipped at room temperature.

 Upon receipt, it should be stored at -20°C.

 Avoid repeated freeze-thaw cycles.

Details

Interleukin 15 Signaling

IL-15 and IL-2 are two closely related cytokines that belong to the IL-2 family and display important functions in the immune system [1]. They share the heterodimeric CD122 (IL-2Rβ)/CD132 (IL-2Rγ) receptor to deliver their signals within target cells. Their specificity of action is conferred by their α receptor chains, CD25 (IL-2Rα) and CD215 (IL-15Rα). The binding of IL-15 and IL-2 to their heterodimeric or heterotrimeric receptors activates a signaling cascade through tyrosine kinases of the Janus family (JAK1 and JAK3) and signal transducer and transcription activator 5 (STAT5) [1]. This results in the subsequent induction of proliferation and anti-apoptotic transcriptional program and can, if uncontrolled, potentially lead to malignant transformation [2]. IL-15 can sustain chronic inflammation, autoimmunity, and uncontrolled T cell proliferation, leading to T cell lymphoma development [1].

IL-15 therapeutic targeting

The multi-faceted nature of IL-15, its anti-tumorigenic abilities as a booster of the immune response, and its pathogenic function as a driver of malignant cell proliferation harbors a high potential as a therapeutic tool [1]. Numerous IL-15-enhancing approaches have been used to treat different cancers, including leukemias and lymphomas, exploiting the main function of the cytokine to potentiate NK and T cell responses and unleash immune-mediated cancer eradication. On the other hand, agents that inhibit IL-15 are also being extensively tested as therapeutic approaches against cancer, such as soluble IL-15Rα, antibodies against IL-15 or components of the heterotrimeric receptor, and steric antagonists of the receptor in the form of a modified IL-2 [1]. Moreover, the IL-15 cytokine, together with IL-2 and IL-7, is currently under investigation for application in ex vivo T cell expansion. These cytokines are used in different concentrations and combinations to improve cell viability, proliferation, and differentiation in CAR T cell culture [2-3]. 

References:

1. Sindaco P, et al., 2023. The role of interleukin-15 in the development and treatment of hematological malignancies. Front Immunol.14:1141208.
2. Sudarsanam H, et al., 2022. Influence of Culture Conditions on Ex Vivo Expansion of T Lymphocytes and Their Function for Therapy: Current Insights and Open Questions. Front Bioeng Biotechnol.;10:886637
3. Chamucero-Millares JA, et al., 2021. Usefulness of IL-21, IL-7, and IL-15 conditioned media for expansion of antigen-specific CD8+ T cells from healthy donor-PBMCs suitable for immunotherapy. Cell Immunol.;360