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 [3]. 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 [3])

• 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.

 Read our mini-review on Immune Checkpoint Blockade.

 Learn more on InvivoGen’s Immune Checkpoint Antibodies.

References:

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.

Figures

Jurkat-Lucia™ NFAT-CD16 cell responses to ADCC induction with anti-human CD20 isotypes and Raji-hCD20 target cells