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Applications: Screen for agonists or antagonists of LIGHT-HVEM signaling in a physiological relevant cellular context Characterize T cell-mediated immune responses of HVEM and its interactions with LIGHT Screen co-stimulatory immune checkpoint molecules
Background: HVEM is a bidirectional switch regulating T-cell activation in a costimulatory or coinhibitory fashion whose outcome depends on the binding partner. HVEM can act as both receptor and ligand, the binding of endogenous ligand LIGHT or agonist antibodies to HVEM delivers a costimulatory signal; whereas the binding of HVEM to BTLA (IgSF) or CD160 on Effector T cells delivers a coinhibitory signal. LIGHT/HVEM axis are co-stimulatory immune checkpoint molecules extensively studied for cancer immunotherapy; LIGHT, either in the soluble monomer form or homotrimer expressed on the cell surface, can activate NF-kB through binding to HVEM; we have shown that LIGHT expressing cells are more potent and efficacious than the soluble LIGHT for activating HVEM expressed on T cell surface.
Description: Recombinant clonal stable Jurkat T cell line expressing firefly luciferase gene under the control of 4 copies of NF-κB response elements with constitutive expression of human HVEM (Herpes Virus Entry Mediator, Tumor Necrosis Factor Receptor Superfamily Member 14, TNFRSF14, CD270, GenBank Accession No. NM_003820). Following activation by human HVEM ligand LIGHT, NF-κB transcription factors bind to the DNA response elements to induce transcription of the luciferase reporter gene.Materials Required but Not Supplied: Assay Medium: Thaw Medium 2 (BPS Cat. #60184)LIGHT, His-Tag (Human) (BPS Cat. #71266LIGHT-CHO Recombinant Cell Line (BPS Cat. # 79262)96-well tissue culture-treated white clear-bottom assay plateOne-Step luciferase assay system (BPS bioscience # 60690) or other luciferase reagents for measuring firefly luciferase activityLuminometer
Host Cell Line: Jurkat
Mycoplasma Testing: This cell line has been screened using the MycoAlert™ Mycoplasma Detection Kit (Lonza, #LT07-118) to confirm the absence of Mycoplasma contamination. MycoAlert Assay Control Set (Lonza, #LT07-518) was used as a positive control.
Storage Stability: Store in liquid nitrogen immediately upon receipt.
Supplied As: Each vial contains ~ 2 x 10^6 cells in 1 ml of 10% DMSO.
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: BSL-1
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