Development of a PD-L1 Overexpression System in HEK293T Cells: A Platform for Studying Immune Evasion Mechanisms
DOI:
https://doi.org/10.61173/enz7xj28Keywords:
PD-L1, immune checkpoint, cancer immunotherapy, tumor microenvironment, overexpression system, HEK293T cellsAbstract
Programmed death-ligand 1 (PD-L1) is a key immune checkpoint protein that facilitates tumor immune evasion by interacting with PD-1 receptors on T cells, making the PD-1/PD-L1 axis a critical target in cancer immunotherapy. However, resistance to these therapies remains a challenge, necessitating further exploration of PD-L1’s functions and regulatory mechanisms. In this study, we amplified the PD-L1 coding sequence (CDS) from human cDNA, cloned it into a pcDNA3.1 vector, and transfected it into HEK293T cells to establish a robust in vitro PD-L1 overexpression system. Quantitative RT-qPCR demonstrated a 300-400 fold increase in PD-L1 mRNA expression compared to controls. These results confirm the successful construction of the PD-L1 overexpression plasmid and validate its application as a tool for studying PD-L1’s role in immune modulation and its potential implications in overcoming resistance to immunotherapy. This system provides a valuable platform for future investigations into PD-L1-mediated mechanisms of immune evasion and cancer progression.
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