1. Jurkat NFAT-GFP Reporter Cell System

Background

The activation of T-cell receptors (TCRs) is a critical component of the adaptive immune response, driven by the interaction between TCRs and peptide-major histocompatibility complex (MHC) molecules on antigen-presenting cells (APCs). Understanding this interaction is pivotal for developing improved checkpoint and cell-based immunotherapies.

To support this research, we have developed a versatile platform utilizing Jurkat NFAT-GFP reporter cells in combination with K562 APCs engineered to express any Class I HLA molecule. This system enables the characterization of antigen specificity and the identification of peptide epitopes recognized by specific TCRs.

Fig 1. Specific TCR-Epitope-MHC interactions activate expression of GFP and secrete IL2 which specifically labels APC expressing interacting peptides. Amplification and NGS analysis from IL2-labelled APCs allows the identification of peptide epitope candidates which are used for follow-up validation and characterization of TCR candidate antigen specificity using synthetic peptides.

Applications of the Jurkat Reporter Cell System:

1. TCR Cloning and Evaluation:
   Jurkat reporter cells facilitate the cloning of full-length TCRs identified through bulk or single-cell Adaptive Immune Receptor Repertoire (scAIR) analysis. This allows for precise determination of the specificity and affinity of both native and engineered TCRs.

2. High-Medium Throughput Antigen Screening:
   The system is ideally suited for medium- to high-throughput screening of peptide libraries. These may include synthetic peptides loaded onto PBMC-derived APCs or expressed in K562 cells as minigene libraries paired with specific HLA Class I molecules.

3. Screening and Evaluation of Novel Therapies
   Jurkat reporter systems are a powerful tool in drug discovery and immunotherapy development. They support the screening of antibodies or small molecules that can mimic or inhibit TCR activation. Hits can be further validated as functional Chimeric Antigen Receptors (CARs).

4. T Cell Signaling Studies
   These cells are widely used to explore TCR signaling pathways. Researchers can use the system to study T-cell responses to tumor- or virus-specific antigens and to dissect the molecular mechanisms underlying T-cell activation.

Please read the entire user manual before proceeding with your experiment.

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