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Application: Positive control for anti-CD19 CAR evaluation in T/NKcells.Transduction optimization experiments.Generate anti-CD19 armored CAR-T/CAR-NK cells (for research use only, not for therapeutic purposes).
Background: CD19 (also known as Cluster of Differentiation 19, B-lymphocyte surface antigen B4, or CVID3) is a glycoprotein expressed at the surface of B lymphocytes through most phases of B cell maturation. It is strictly required for B cell terminal differentiation. Mutations in the CD19 gene cause severe immune-deficiency syndromes associated with impaired antibody production such as CVID3 (common variable immuno-deficiency 3). The majority of B cell malignancies express normal to high levels of CD19, making it a nearly ideal target for cancer immunotherapy. Blinatumomab, a CD19/CD3 bi-specific T cell engager (BiTE) has been approved for relapsed/refractory B precursor ALL (Acute lymphoblastic leukemia) and CD19 was the target of the first approved CAR-T cell therapy. Recently, a new generation of CAR-T cells has been developed. These 4th generation CAR-T constructs include a ligand or cytokine, with the goal of armoring the T cells against the TME (tumor microenvironment). The addition of IL-15 has been shown to increase the cytotoxic potential of CAR-T cells both in vitro and in vivo. The continuous improvement of CAR-T cells will lead to more efficacious cancer therapies. Moreover, it has been reported that engineering NK cells with the IL-15/IL-15R can further improve NK cell proliferation, activation, and cytotoxic activity against B-cell leukemiaα.
Description: The Anti-CD19 CAR Lentivirus (CD19 ScFv-CD8-CD28-CD3ζ, IL-15/IL-15Ra) are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to transduce almost all types of mammalian cells, including primary and non-dividing cells. These viruses transduce the ScFv portion of anti-CD19 (clone FMC63) linked to a 4th generation CAR (Chimeric Antigen Receptor) containing the CD8 hinge and transmembrane domains, CD28 and CD3ζ signaling domains. This construct also includes a 2A-IL15/IL-15 receptor alpha (IL15Ra) sequence downstream of the anti-CD19 CAR cassette to create armored CAR-T cells by overexpressing IL-I5 and IL-15Ra.Note: This product transduces the same construct as the anti-CD19 CAR Lentiviruses (CD19 ScFv-CD8-4-1BB-CD3ζ, IL-15/IL-15Ra) (#82378), but differs in a key aspect:82379, described here, is constructed with a CD28 costimulatory domain in the anti-CD19 CAR cassette, while 82738 contains a 4-1BB costimulatory domain.
Formulation: The lentiviruses were produced from HEK293T cells, concentrated, and resuspended in DMEM. Virus particles can be packaged in custom formulations by special request, for an additional fee.
Storage Stability: Lentiviruses are shipped with dry ice. For long term storage, it is recommended to store the virus at -80°C for up to 12 months from date of receipt. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.
Supplied As: 50 µl at a titer ≥ 3 x 108 TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.
Warnings: Avoid freeze/thaw cycles
Biosafety Level: BSL-2
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