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Scientific 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.
Product Description
HIV-based · VSV-G-pseudotyped · SIN=YesIL15/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. IL15 and IL15Ra are fused via a glycine-serine linker.
Technical Details
| Vector Type | HIV-based, VSV-G-pseudotyped lentiviral vector |
|---|---|
| Payload / Construct | IL15/IL15Ra fusion transgene |
| Target Antigen / Gene | IL-15 / IL-15Rα (IL15, IL15RA) |
| Signaling / Architecture | N/A (transgene expression) |
| Selection Marker | Puromycin |
| Reporter | None |
| Biosafety Level | BSL-2 |
| SIN Vector | Yes |
| Formulation | The lentivirus particles were produced in HEK293T cells in medium containing 90% DMEM + 10% FBS. |
| 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. |
| Storage | −80°C; avoid repeated freeze-thaw cycles |
| Hazardous Shipping | UN3373 |
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
Biosafety & Safety
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, bpsbioscience.com/cell-line-faq), for antibiotic selection of transduced cells, followed by clonal selection. 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 support@bpsbioscience.com.
To generate a IL-15 / IL-15Rα-expressing stable cell line, transduce your cells of interest at an appropriate MOI. Remove growth medium 48 hours post-transduction and replace with fresh medium containing puromycin at a concentration pre-determined by a kill curve. Continue selection until untransduced control cells are fully eliminated.
BSL-2 facilities are required. The lentiviruses are produced using a SIN (self-inactivating) lentivector design, ensuring self-inactivation after integration. HIV accessory genes are not expressed in transduced cells. Always follow applicable institutional and regulatory biosafety guidelines.
Each shipment contains 500 µl × 2 of lentivirus at ≥10⁷ TU/ml. The exact titer is lot-specific and is reported on the Certificate of Analysis provided with each shipment.
This lentivirus encodes a puromycin resistance gene. Following transduction, stable integrants can be selected by puromycin treatment. Determine the optimal puromycin concentration for your cell line using a kill-curve assay prior to selection.
This VSV-G-pseudotyped lentiviral vector has broad tropism and can transduce most mammalian cell types, including primary cells, non-dividing cells, and established cell lines. Transduction efficiency may vary by cell type and MOI.
Store at −80°C immediately upon receipt. Avoid repeated freeze-thaw cycles to preserve titer. Aliquot into single-use volumes before freezing for best results.
BioHippo offers customization and add-on services for selected products. Options may include:
- Custom formulations: Virus particles can be packaged in alternative formulations upon request (additional fees may apply).
- Custom titers: Higher-titer preparations may be available; contact us for feasibility and pricing.
- Bulk orders: Volume discounts are available for qualifying order quantities.
- Technical support: Our scientific team can assist with protocol optimization and troubleshooting.
For all customization inquiries, please use the contact form or email support@biohippo.com.
- 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.
