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Applications: Validate different CAR designs and constructs for their specificity, efficacy and potency before proceeding into patient-derived primary T cells.
Intracellular co-stimulatory and activation domain comparison.
Background: The development of CAR-T cells is a complex process that requires multiple steps in the workflow including I) screening and sequencing of mAbs that are specific to the cancer antigens; II) engineering and validation of scFv and scFv-CAR of different varieties for their specificities and activities; III) production of high titer lentivirus for CAR constructs; IV) isolation, activation and expansion of primary T cells from healthy donors or patients that exhibit a specific cellular phenotype; V) transduction of activated T cells with CAR-encoding lentivirus; VI) validation of engineered CAR-T cells through FACS and functional analysis. BPS Bioscience has developed a series of CAR-T products, including lentiviruses, reporter cell lines and fully validated functional CAR-T cells for a variety of targets such as CD19 and BCMA. In this product, anti-CD19 CAR negative control and NFAT-luciferase reporter are co-transfected into a Jurkat cell line, where the anti-CD19 scFv binds to CD19, however, it does not induce the activation of CAR and luciferase reporter through NFAT as the intracellular activation motifs are missing. Anti-CD19 scFv linked to the CD28 transmembrane region was cloned into a lentivector, and packaged using a safe, replication incompetent, and VSV-G pseudotyped lentiviral packaging system, in which the gene of anti-CD19 CAR negative control is driven by an EF-1α promotor. Anti-CD19 CAR negative control Jurkat/NFAT reporter cell line was generated by the transduction of anti-CD19 CAR negative control lentivirus into an NFAT-luciferase reporter Jurkat cell line. In these cells, the luciferase reporter should not be activated upon co-culture with CD19/CHO target cells (BPS Bioscience #79561).Anti-CD19 CAR/NFAT-luciferase reporter Jurkat cell line (BPS Bioscience, #79853) is a great system for primary screening of anti-CD19 CAR and predicting its mechanism of action before testing on patient-derived primary T cells. The same anti-CD19 CAR lentivirus (BPS Bioscience, #79851) was also used to transduce primary T cells to make primary anti-CD19 CAR-T cells, which showed IFN-γ production and cytotoxic killing of CD19+ tumor cells in co-culture experiments, indicating that there is a good correlation between the reporter activity in CAR reporter Jurkat cell line and functional activation of primary CAR-T cells when co-cultured with target cells.
Description: Anti-CD19 CAR negative control/NFAT-luciferase reporter Jurkat cell line is a double stable cell line expressing anti-CD19 CAR negative control and NFAT-luciferase reporter. The anti-CD19 CAR negative control consists of anti-CD19 scFv linked to the CD28 transmembrane motif without any intracellular signaling domains. The reporter cell line has been validated for anti- CD19 expression by FACS, while the stimulation by target cells including CD19/CHO recombinant cell line has not activated the luciferase reporter gene in this cell line. The cell line can be used for the negative control of anti-CD19 CAR/Jurkat-NFAT cell line (BPS Bioscience, #79853).Â
Host Cell Line: Jurkat
Mycoplasma Testing: The cell line has been screened using Lonza MycoAlert Mycoplasma Detection kit (Lonza, #LT07-318) to confirm the absence of Mycoplasma species.
Storage Stability: Immediately upon receipt, store in liquid nitrogen.
Supplied As: Each vial contains 2 x 10^6 cells in 1 ml of 10% DMSO and 90% FBS
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-1
References: 1. Immune checkpoint blockade and CAR-T cell therapy in hematologic malignancies.Wang et al. J Hematol Oncol. 2019 Jun 11;12(1):59-78.
2. Chimeric antigen receptor T cell therapy for multiple myeloma. Hasegawa et al. Inflamm Regen. 2019 Jun 4;39:10-14.
3. Novel targets for the treatment of relapsing multiple myeloma. Giuliani et al. Expert Rev Hematol. 2019 Jun 3:1-16.
4. Anti-CD19 antibodies in the future management of multiple myeloma. Gavriatopoulou et al. Expert Rev Anticancer Ther. 2019 Apr;19(4):319-326.
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