Related Experiment Video
Updated: Jan 16, 2026
Author Spotlight: High-Throughput Screening of CAR T-Cell Constructs for Enhanced Cytotoxicity and Immunologic Memory
Published on: October 27, 2023
Evaluation of Immune Checkpoint Inhibitors in a Jurkat-Based Transcriptional Reporter System: A Cell-Based Reporter
Magdalena Bojko1, Judith Leitner2, Peter Steinberger2
1Department of Biomedical Chemistry, Faculty of Chemistry, University of Gdańsk, Gdańsk, Poland. magdalena.bojko@phdstud.ug.edu.pl.
Abstract:
Humanity has been facing cancer since the beginning of its existence, and the number of new cancer cases grows each year. Immunotherapies based on immune checkpoint inhibition have been intensively developed over the last decade and focus on the blockade of the co-inhibitory molecule complexes such as CTLA-4 with its ligands CD80 and CD86, PD-1 with PD-L1 and PD-L2, and many more. The investigation and development of new immune checkpoints inhibitors is necessary for better cancer treatment. One way of inhibitor evaluation is to assess them in cell-based assays. In this protocol, we focus on describing the cell-based reporter platform for PD-1/PD-L1 inhibitors assessment, which is based on measuring the expression of eGFP under the transcription factor NF-κB, responsible for transcriptional program required for T-cell activation and differentiation.
Insights
This study presents a novel cell-based reporter assay for evaluating programmed cell death protein 1 (PD-1) and its ligand 1 (PD-L1) inhibitors. This method aids in developing new cancer immunotherapies by assessing inhibitor efficacy through NF-κB activation.
Area of Science:
- Oncology
- Immunology
- Biotechnology
Background:
- Cancer incidence is rising globally, necessitating advanced treatment strategies.
- Immune checkpoint inhibitors, targeting complexes like PD-1/PD-L1, are a key focus in modern cancer therapy.
- Developing novel inhibitors requires robust evaluation methods.
Purpose of the Study:
- To describe a new cell-based reporter platform for assessing PD-1/PD-L1 inhibitors.
- To provide a method for evaluating the efficacy of immune checkpoint inhibitors.
Main Methods:
- Development of a cell-based reporter assay.
- Utilizing the expression of enhanced green fluorescent protein (eGFP) as a readout.
- Measuring eGFP expression under the control of the NF-κB transcription factor.
Main Results:
- The reporter platform enables the assessment of PD-1/PD-L1 inhibitor activity.
- NF-κB activation, indicative of T-cell activation, is measured via eGFP expression.
- This assay provides a quantifiable method for inhibitor screening.
Conclusions:
- The described reporter platform is a valuable tool for evaluating PD-1/PD-L1 inhibitors.
- This assay facilitates the development of more effective cancer immunotherapies.
- The NF-κB-driven eGFP reporter system aids in understanding T-cell activation pathways relevant to immunotherapy.
More Related Videos
14:27CRISPR-Cas9-based Genome Engineering to Generate Jurkat Reporter Models for HIV-1 Infection with Selected Proviral Integration Sites
Published on: November 14, 2018
05:21Author Spotlight: Optimizing Antibody-Based Cancer Treatments via Antibody-Dependent, Cell-Mediated Cytotoxicity Assay
Published on: September 13, 2024