TIM3/NFκB Reporter Lentivirus

Background

TIM-3 (HAVCR2) is a co-inhibitory receptor expressed on T cells, natural killer cells, and myeloid cells, where it acts as a checkpoint that limits immune activation. Engagement by ligands such as galectin-9, phosphatidylserine, CEACAM1, and HMGB1 contributes to the inhibition of effector responses and is a hallmark of T cell exhaustion in chronic infection and cancer. TIM-3 modulates downstream signaling pathways, including the NF-κB family of transcription factors that govern immune activation and inflammatory gene expression. Because of its role in restraining anti-tumor immunity, TIM-3 is a prominent target for checkpoint inhibitor therapies, and pathway reporters help quantify its signaling.

Product Description & Applications

The TIM3/NF-κB Reporter Lentivirus is an immunotherapy reporter system for studying the checkpoint receptor TIM-3 and its NF-κB pathway relevance. The system drives NF-κB-dependent transcription of dual reporters: secreted Gaussia luciferase and a fluorescent protein (GFP or RFP), enabling quantification of NF-κB activation and real-time visualization of pathway-specific effects.

Sequential transduction and selection generate a stable, pathway-specific reporter cell line. Applications include studying the role of TIM-3 in cancer immunity, T cell exhaustion, and checkpoint inhibitor therapies. Supplied as third-generation, VSV-G-pseudotyped lentiviral particles purified by PEG precipitation and sucrose gradient centrifugation, the product is effective in difficult-to-transfect primary and thawed cells.

About This Product

This 2-vial immunotherapy reporter system consists of a Vial 1 Receptor Lentivirus encoding human TIM3 under a constitutive promoter with antibiotic selection, and a Vial 2 Reporter Lentivirus encoding tandem NFAT (or NF-κB) response elements driving a dual reporter (GFP, GFP-P2A-GLuc, GLuc, GLuc-P2A-GFP, GLuc-P2A-RFP, RFP, RFP-P2A-GLuc). Sequential transduction and selection generates a dual-stable effector cell line that responds quantitatively to receptor stimulation with a ratiometric fluorescent + bioluminescent readout.

Secreted Gaussia luciferase (where included) accumulates in conditioned media, enabling kinetic sampling without cell lysis. The combined fluorescent and luminescent outputs allow parallel microscopy-based visualization and plate-reader luminometry from the same cell population — providing assay redundancy and flexibility for potency testing formats compliant with regulatory expectations for cell-based functional assays.