Raji-hCTLA4 Cells [NEW]
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Human lymphoblast cells - ADCC CTLA-4 Target Cells
3-7 x 10e6 cells
Human CTLA-4-expressing B-cell line
Raji-hCTLA4 cells were developed from the human Raji cell line. Raji cells have been successfully used as target cells in human effector studies such as antibody-dependent cellular cytotoxicity (ADCC), either with peripheral blood mononuclear cells, Natural Killer cells, or Jurkat-derived reporter T cells. CTLA-4 (cytotoxic T cell lymphocyte antigen-4; also known as CD152) is a type I transmembrane protein expressed at the cell surface of activated conventional T cells and constitutively on immunosuppressive regulatory T cells (Tregs). CTLA-4 is an inhibitory immune checkpoint that prevents T-cell overstimulation and host damage. It exerts competitive binding for stimulatory CD28 ligands (CD80/CD86) .
Features of Raji-hCTLA4 cells:
- Stable overexpression of the human CTLA-4 gene
- Constitutive expression of human CD19 and human CD20 at the cell surface
- Constitutive low expression of human PD-L1 at the cell surface
- Stable expression of a resistance gene to Blasticidin
Applications for Raji-hCTLA4 cells:
- ADCC target cells for anti-hCTLA4, anti-hCD19, anti-hCD20, or anti-hPD-L1 Abs
- Negative control target cells in ADCC assays using Abs other than anti-hCTLA-4, anti-hPD-L1, anti-hCD19, or anti-hCD20 Abs
- Target cells for anti-hCD19-CD3 bispecific Abs
Validation of Raji-hCTLA4 cells:
- Human CTLA-4 expression verified by flow-cytometry
- Functionally tested as target cells in ADCC assays using anti-human CTLA-4 monoclonal Abs and Jurkat-Lucia™ NFAT-CD16 cells
1. Ribas A. and Wolchock J.D., 2018. Cancer immunotherapy using checkpoint blockade. Science. 359:1350-55.
Comparison of ADCC potency for native and engineered anti-human CTLA-4 antibody isotypes: Raji-hCTLA4 cells were incubated with gradient concentrations of Anti-hCTLA4 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.
- Human CTLA-4 expression has been verified by flow-cytometry.
- Induction of antibody-dependent cellular cytotoxicity (ADCC) has been validated using InvivoGen’s anti-hCTLA4-hIgG1 and Jurkat-NFAT Lucia™ CD16 reporter cell line.
- The stability for 20 passages following thawing has been verified.
- Raji-hCTLA4 cells are guaranteed mycoplasma-free.
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- 1 vial of Raji-hCTLA4 cells (3-7 x 106 cells) in Freezing Medium
- 1 ml of Blasticidin (10 mg/ml). Store at 4 °C or at -20 °C.
- 1 ml of Normocin™ (50 mg/ml). Normocin™ is a formulation of three antibiotics active against mycoplasmas, bacteria and fungi. Store at -20 °C.
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