Jurkat-Lucia™ NFAT Cells for ADCC & ADCP Assays
Jurkat-Lucia™ NFAT-CD16 Cells | Unit size | Cat. code | Docs | Qty | Price |
---|---|---|---|---|---|
ADCC Reporter Cells - Human T Lymphocytes |
3-7 x 10e6 cells |
jktl-nfat-cd16 |
Jurkat-Lucia™ NFAT-CD32 Cells | Unit size | Cat. code | Docs | Qty | Price |
---|---|---|---|---|---|
ADCP Reporter Cells - Human T Lymphocytes |
3-7 x 10e6 cells |
jktl-nfat-cd32 |
You may also need : Zeocin™ | View more associated products ▼
ADCC and ADCP reporter T-cell lines
Intracellular signaling in Jurkat™-Lucia NFAT-CD16 (or CD32) cells
InvivoGen also offers:
• Clinically relevant antibodies in various Ig isotypes
• Raji-target cells expressing a choice of molecules of interest
InvivoGen offers a series of Jurkat-Lucia™ NFAT cell lines, specifically designed to assess the potency of specific immunoglobulin for ADCC (antibody-dependent cellular cytotoxicity) and ADCP (antibody-dependent cell-mediated phagocytosis).
– Jurkat-Lucia™ NFAT-CD16 cells for ADCC reporter assays
– Jurkat-Lucia™ NFAT-CD32 cells for ADCP reporter assays
These cells derive from the human T-lymphocyte Jurkat cell line and stably express CD16 or CD32, two Fc-gamma receptors (FcγR) for the constant region of immunoglobulins. ADCC and ADCP outcomes depend on the preferential affinity of each antibody (Ab) for one FcγR. The Ab-FcγR interaction is regulated by the Ab isotype and glycosylation. Jurkat cells naturally express a functional NFAT (nuclear factor of activated T cells) transcription factor, which is involved in the early signaling events of ADCC and ADCP [1, 2].
Jurkat-Lucia™ NFAT-CD16 and Jurkat-Lucia™ NFAT-CD32 cells have been designed as effector reporter cells for InvivoGen’s ADCC and ADCP assays, respectively. These cells stably express the Lucia luciferase reporter gene under the control of an ISG54 minimal promoter fused to six NFAT response elements. ADCC or ADCP induction is measured as a bioluminescent signal produced by the Lucia luciferase upon the addition of the appropriate detection reagent QUANTI-Luc™.
Key Features:
- Endogenous NFAT expression
- Stable CD16A(FcgRIIIA; V158 allotype [3]) expression
- Stable CD32A(FcgRIIA; H131 allotype [4])expression
- Readily assessable Lucia luciferase reporter activity for NFAT activation
- Resistance to Blasticidin and Zeocin™
Applications:
- Screening of engineered monoclonal antibodies (mAbs) potency for ADCC using Jurkat-Lucia™ NFAT-CD16 cells
-
Screening of engineered mAbs potency for ADCP using Jurkat-Lucia™ NFAT-CD32 cells
Read our review on Immune Checkpoint Blockade.
Learn more about 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 activation: 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.
4. Tay M.Z. et al.., 2019. Antibody-Dependent Cellular Phagocytosis in Antiviral Immune Responses. Front Immunol. 10:332.
Specifications
Growth medium: IMDM, 2 mM L-glutamine, 25 mM HEPES, 10% (v/v) heat-inactivated fetal bovine serum (FBS), 100 U/ml penicillin, 100 µg/ml streptomycin, 100 µg/ml Normocin™
Antibiotic resistance: Blasticidin and Zeocin™
Quality Control:
- Human CD16A and CD32A expression have been verified by flow-cytometry.
- Induction of antibody-dependent cellular cytotoxicity (ADCC) and antibody-dependent cellular phagocytosis (ADCP) have been validated using InvivoGen’s anti-hCD20-hIgG1, anti-hPD-L1-hIgG2, Raji-Null cells, and Raji-hPD-L1 cells.
- The stability for 20 passages following thawing has been verified.
- These cells are guaranteed mycoplasma-free.
These products are covered by a Limited Use License (See Terms and Conditions).
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Please note: Each cell line is sold separately. See TDS for the exact contents of each cell line.
- 3-7 x 106 Jurkat-Lucia™ NFAT-CD16 cells OR Jurkat-Lucia™ NFAT-CD32 cells in a cryovial or shipping flask
- 1 ml of Blasticidin (10 mg/ml)
- 1 ml of Zeocin™ (100 mg/ml)
- 1 ml of Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasmas, bacteria, and fungi.
- 1 pouch of QUANTI-Luc™
Shipped on dry ice (Europe, USA & Canada).
Details
ADCC and ADCP are immune mechanisms through which Fc receptor-bearing effector cells can recognize and clear antibody (Ab)-coated microbes and target cells expressing specific antigens on their surface. The binding of antibody-antigen complexes to activatory FcγRs induces their cross-linking and subsequent signaling through immunoreceptor tyrosine-based activation motifs (ITAMs). Cytoplasmic signaling includes an increase in intracellular calcium concentration and calcineurin/calmodulin-mediated dephosphorylation of NFAT (nuclear factor of activated T cells), allowing its nuclear translocation and binding to promoter regions of ADCC and ADCP relevant genes [1, 2].
The ADCC and ADCP outcomes depend on the preferential affinity of each Ab for one FcgR. The Ab-FcgR interaction is regulated by the Ab isotype and glycosylation.
Antibody-dependent cellular cytotoxicity (ADCC)
ADCC is triggered by the cross-linking between antigen-bound Abs and the Fc receptor CD16A (FcγRIIIA)at the surface of Natural Killer (NK) cells [3]. Activated NK cells release cytotoxic granules which kill the microbe or target cells [1].
Antibody-dependent cell-mediated phagocytosis (ADCP)
ADCP is triggered by the cross-linking between antigen-bound Abs and the Fc receptor CD32A (FcγRIIA) at the surface of monocytes, macrophages, and dendritic cells. These interactions induce the phagocytosis of the microbe or target cells. This internalization is followed by phagolysosomal degradation, thus facilitating antigen presentation and stimulating inflammatory cytokine secretion [2].
References:
1. Quast I. et al., 2016. Regulation of antibody effector functions through IgG Fc N-glycosylation. Cell. Mol. Life. Sci. 74(5):837-47.
2. Tay M.Z. et al., 2019. Antibody-Dependent Cellular Phagocytosis in Antiviral Immune Responses. Front Immunol. 10:332