CD27/NFκB Reporter Lentivirus

Background

CD27 is a co-stimulatory receptor of the tumor necrosis factor receptor superfamily, expressed on T cells, natural killer cells, and subsets of B cells. Engagement by its ligand CD70 promotes T cell activation, proliferation, survival, and the generation of effector and memory populations. CD27 signaling proceeds through TRAF adaptor proteins to activate the NF-κB pathway, a family of transcription factors that controls expression of genes governing immune activation, survival, and inflammation. Because CD27-CD70 co-stimulation amplifies anti-tumor and anti-viral immunity, the axis is an active target for agonist immunotherapies, and NF-κB-driven reporters provide a quantitative measure of CD27 pathway engagement.

Product Description & Applications

The CD27/NF-κB Reporter Lentivirus is an all-in-one immunotherapy reporter system for studying CD27 signaling in T cells. The construct constitutively expresses the human CD27 receptor and couples receptor engagement to NF-κB-dependent transcription of dual reporters: secreted Gaussia luciferase and a fluorescent protein (GFP or RFP). This design enables real-time monitoring of CD27-mediated immune activation by both luminometry and fluorescence.

Transducing target cells with the high-titer lentivirus establishes a stable, pathway-specific reporter cell line. Applications include evaluating co-stimulatory signals, studying immune responses, and testing CD27-targeted immunotherapeutics. Supplied as third-generation, VSV-G-pseudotyped particles purified by PEG precipitation and sucrose gradient centrifugation, the product performs well 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 CD27 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.