IL15/IL15Ra Lentivirus

Catalog #

78938

$1,250 *

Size: 500 µl x 2

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Description

IL15/IL15Ra Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles ready to transduce nearly all types of mammalian cells, including primary and non-dividing cells. These particles result in expression of human IL15 (NM_000585.4) and IL15Rα (NM_002189.3) driven by an EF1a promoter, and a puromycin selection marker (Figure 1). IL15 and IL15Ra are fused via a glycine-serine linker.

Figure 1. Schematic of the lenti-vector used to generate IL15/IL15Ra Lentivirus.

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Product Data Gallery

Expression of IL15 and IL15Ra in CHO cells transduced with IL15/IL15Ra Lentivirus

Product Info

Storage and Usage

Citations

Supplied As

Two vials (500 µl x 2) of lentivirus at a titer ≥10⁷ TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.

Formulation

The lentivirus particles were produced in HEK293T cells in medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations and produced at higher titers by special request, for an additional fee.

Background

IL15 (interleukin 15) and its high affinity receptor (IL15R) are involved in NK cell development and proliferation and persistence of CD8⁺ T cells, NKT cells, δγT cells and NK cells. The use of membrane bound IL15 as part of the CAR in T cells has been in use for a few years, as a way to improve T cell persistence in vivo. IL15 has a short half-life and requires treatment in high dosages, so research into ways to increase its potency has been ongoing. RLI-15 (receptor-linker-IL-15) is considered a super agonist composed of IL15 and an IL15Rα domain. This fusion protein was designed to bypass the need for endogenous IL15Rα to take advantage of IL15. It has been shown to activate proliferation and activity of NK. RLI-15 resulted in a potent effect in cell models. Recently the use of a CAR containing IL15/IL15Rα and targeting CD19 in NK92 cells has also resulted in increased proliferation, cytokine secretion and cytotoxicity towards B-cell cancer cell lines. The inclusion of IL15/IL15Rα fusion proteins to armor CARs represents a step forward in the fight against solid tumors.

References

Hurton L., et al., 2016 Proc Natl Acad Sci USA 113 (48): E7788-E7797.
Fujii R., et al., 2018 Cancer Immunol Immunother. 67(4):675-689.
Desbois M., et al., 2020 Journal for ImmunoTherapy of Cancer 8:e000632.
Silvestre R., et al., 2023 Front. Immunol. 14: https://doi.org/10.3389/fimmu.2023.1226518.

Storage/Stability

Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses at -80°C. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.

Applications

Expression of human IL15/IL15Ra in cells of interest.
Arming CAR-T and CAR-NK cells with IL15/IL15Ra fusion protein.
Generate cell pools or stable cell lines expressing IL15/IL15Ra following puromycin selection

Shipping Temperature

-80°C

Notes

To generate an IL15/IL15Ra expressing stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin (as pre-determined from a killing curve, https://bpsbioscience.com/cell-line-faq), for antibiotic selection of transduced cells, followed by clonal selection.

Biosafety
The lentiviruses are produced with a SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and after integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal and are not present in the lentivirus particle. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.

Troubleshooting Guide
Visit bpsbioscience.com/lentivirus-faq for detailed troubleshooting instructions. For further questions, please email [email protected].

Warnings

Avoid freeze/thaw cycles

Scientific Category

Immunotherapy, NK Cells

Regulatory Status

Research Use Only

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IL15/IL15Ra Lentivirus

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