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Applications: The CD40 NF-κB Reporter HEK293 Stable Cell Line is ideal for High Throughput Screening (HTS) to identify potential CD40 agonistic monoclonal antibodies and CD40-specific inhibitors.
Background: CD40, a TNF receptor superfamily member, was initially identified on B lymphocytes. Antigen presenting cells (APCs) such as monocytes, basophils, dendritic cells, have been found to express CD40. Additionally, CD40 has been identified in non-immune cells such as endothelial cells and epithelial cells. A wide variety of carcinoma cells over-express CD40 as well. Interaction with CD40 ligand (CD40L, CD154) on CD4+ T helper lymphocytes triggers the expression of intercellular adhesion molecule (ICAM) and other pro-inflammatory cytokines. CD40:CD40L signaling simultaneously increases activation of antigen-specific T cells. CD40 activates NF-κB-dependent signaling in response to lipopolysaccharide (LPS) found on Gram negative bacterial pathogens. Furthermore, agonistic CD40 monoclonal antibodies have been shown to activate antigen presenting cells (APC) and promote anti-tumor T-cell responses in addition to fostering cytotoxic myeloid cells with the potential to control cancer in the absence of T-cell immunity.
Description: Recombinant HEK293 cell line expressing full length human CD40 (Tumor necrosis factor receptor superfamily member 5; TNFRSF5). Expression is confirmed by flow cytometry and Western Blot. This NF-κB luciferase reporter construct is stably integrated into the genome. The firefly luciferase gene is controlled by 4 copies of NF-κB response element located upstream of the TATA promoter. Following activation by human CD40 ligand, NF-κB transcription factor binds to the DNA response elements to induce transcription of the luciferase gene.
Format: Each vial contains ~2 x 106 cells in 1mL of 10% DMSO in FBS.
Host Cell Line: HEK293
Mycoplasma Testing: This cell line has been screened using the MycoAlert™ Mycoplasma Detection Kit (Lonza, Cat.#LT07-118) to confirm the absence of Mycoplasma contamination. MycoAlert Assay Control Set(Lonza, Cat. #LT07-518) was used as a positive control.
Storage Stability: Immediately upon receipt, store vials in liquid nitrogen.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-2
References: 1.Li G et.al. (2013) Human Genetics in Rheumatoid Arthritis Guides a High-Throughput Drug Screen of the CD40 Signaling Pathway. PLoS Genet. 9: e1003487. 2.Pontrelli P et.al. (2006) CD40L Proinflammatory and Profibrotic Effects on Proximal Tubular Epithelial Cells: Role NF-κB and Lyn. J. Amer. Soc. Neph. 17: 627. 3.Lavorgna A et.al. (2014) A Critical Role for IL-17RB Signaling in HTLV-1 Tax-Induced NF-κB Activation and T-Cell Transformation. PLoS Path. 10: e1004418.4.Moschonas A et.al. (2012) CD40 Stimulates a "Feed-Forward" NF-κB- Driven Molecular Pathway That Regulates IFN-β Expression in Carcinoma Cells. J. Immunol. 188: 5521.
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