Jurkat-Lucia™ NFAT-CD16 Cells
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Human T Lymphocytes - ADCC Reporter Cells
3-7 x 10e6 cells
ADCC reporter T-cell line
Jurkat-Lucia™ NFAT-CD16 cells were engineered from the human T-Lymphocyte Jurkat cell line. Jurkat cells naturally express a functional NFAT (nuclear factor of activated T cells) transcription factor, which is involved in the early signaling events in antibody-dependent cellular cytotoxicity (ADCC) [1, 2]. ADCC is an immune mechanism through which Fc receptor-bearing effector cells can recognize and kill antibody (Ab)-coated target cells expressing antigens on their surface. ADCC is triggered by the cross-linking between antigen-bound Abs and the Fc receptor CD16A at the surface of immune effector cells. These interactions induce the increase of intracellular calcium concentrations, calicneurin/calmodulin-mediated dephosphorylation of NFAT, allowing its nuclear translocation and binding to promoter regions of ADCC relevant genes. Ultimately, the effector cells release cytotoxic granules which kill the target cells . Jurkat-Lucia™ NFAT-CD16 cells have been designed as effector reporter cells for InvivoGen’s ADCC assay. These cells stably express the Lucia luciferase reporter gene under the control of an ISG54 minimal promoter fused to six NFAT response elements. ADCC induction is measured as a bioluminescent signal produced by the Lucia luciferase upon addition of the appropriate detection reagent QUANTI-Luc™.
Features of Jurkat-Lucia™ NFAT-CD16 cells:
- Stable expression of the cell surface Fc receptor CD16A (FcgRIIIA; V158 high affinity allotype )
- Stable expression of the Lucia luciferase reporter gene under the control of an ISG54 minimal promoter fused to six NFAT response elements
- Resistance to Blasticidin and Zeocin™
Applications for Jurkat-Lucia™ NFAT-CD16 cells:
- ADCC reporter effector cells
Validation of Jurkat-Lucia™ NFAT-CD16 cells:
- Human CD16A expression verified by flow-cytometry
- Functionally tested in ADCC assays with various target cells from our expanding collection of Raji-derived target cells (e.g. Raji-Null cells, Raji-hCTLA4 cells, Raji-hPD-1 cells, Raji-hPD-L1 cells) and specific monoclonal Ab isotype combinations, such as immune checkpoint antibodies.
Learn more on InvivoGen’s Immune Checkpoint Antibodies.
1. Shaw J-P. et al., 1998. Identification of a putative regulator of early T cell activation genes. Science. 241:202-205.
2. Leibson P.J., 1997. Signal transduction during natural killer cell activaion: inside the mind of a killer. Immunity. 6:655-61.
3. Quast I. et al., 2016. Regulation of antibody effector functions through IgG Fc N-glycosylation. Cell. Mol. Life. Sci. 74(5):837-47.
Comparison of ADCC potency for native and engineered anti-human CD20 isotypes: Raji-Null cells were incubated with gradient concentrations of Anti-hCD20 or Anti-β-galactosidase (β-gal) mAbs for 1 hour. Jurkat-Lucia™ NFAT-CD16 effector cells were then co-incubated with targets cells for 6 hours. NFAT activation, reflecting the induced ADCC response, was assessed by determining Lucia luciferase activity in the supernatant using QUANTI-Luc™. Percentages of the maximal response normalized to the IgG1 isotype are shown.
Increased ADCC activity mediated by IgG1 compared to IgG1fut (non-fucosylated): Raji-hCTLA4, Raji-hPD-1, and Raji-hPD-L1 cells were incubated with Jurkat-Lucia™ NFAT-CD16 effector cells and corresponding IgG1 or IgG1fut specific mAbs. The data represent the EC50 for each antibody.
- Human CD16A expression has been verified by flow-cytometry.
- Induction of antibody-dependent cellular cytotoxicity (ADCC) has been validated using InvivoGen’s anti-hCD20-hIgG1 antibody and Raji-Null target cell line.
- The stability for 20 passages following thawing has been verified.
- Jurkat-Lucia™ NFAT-CD16 cells are guaranteed mycoplasma-free.
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- 1 vial of Jurkat-Lucia™ NFAT-CD16 cells (3-7 x 106 cells)
- 1 ml of Blasticidin (10 mg/ml)
- 1 ml of Zeocin™ (100 mg/ml)
- 1 ml of Normocin™ (50 mg/ml)
- 1 pouch of QUANTI-Luc™
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