Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69 upregulation in 6h, I could not find Raji killing by this CAR-Jurkat cells. It is really confusing. I wonder whether it is related to CAR affinity or immune synpase.
In my experience Jurkat cells aren't really efficient for CAR-mediated killing. I don't know why they worked against the Rajis, maybe just because anti-CD19 CARs and Raji cells are an extremely potent combination. Still, Jurkats aren't CD8+, but more like CD4+ T cells. You can stimulate them to produce cytokines like IL-2 and IFN-gamma, which can be indicative for would-be-efficient cytotoxicity, but eventually you'll have to generate primary CAR-T cells to really test the CAR's potency. Hope this helps.
you have to use primary T cells. Jurkat cells dont have sufficient cytotoxicity to kill Cd19 positive cells although they upregulate activation markers
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