Sigma-Aldrich Bw5147 Murine Thymoma Pd-L1 Cell Line

Immune checkpoints, such as programmed cell dead protein-1 (PD-1) and B and T cell attenuator (BTLA), have become important targets in cancer immunotherapy. These checkpoints limit antitumor immunity so blocking them can result in improved antitumor cell responses. Many modern clinical trials have tested PD-1 or PD-1 ligand (PD-L1) antibody treatments in combination with other cancer therapies to improve outcomes. While benefits are limited to certain patient subsets, immune checkpoint inhibitor (ICI) therapy appears to have clear positive outcomes.1

While BTLA appears to have some biological similarities to PD-1, it is still incompletely understood. Combining PD-1 treatments with BTLA treatments appears feasible but requires more thorough investigation. TCR induced gene expression is an important subject to consider when understanding the function of immune checkpoints. In addition, data reporting how co-inhibitory receptor binding affects gene expression activated by TCR complex signaling is scarce.

The Jurkat JE6.1 NF-κB cell line expressing PD-1 (SCC676) expresses an NF-κB::eGFP reporter construct, resulting in downstream eGFP expression when TCR activation occurs. These cells were designed to be used in conjunction with the BW5147 Murine Thymoma T Cell Stimulator (TCS) cell line expressing PD-L1 (SCC675) as a co-culture system. The BW5147 cells express membrane bound anti-CD3 single chain antibody fragments fused to human CD14 in addition to the costimulatory ligand CD86 used to bind to TCR complexes and activate T-cells. The BW5147 cells also express PD-L1 which can be used to further understand transcriptomic changes that occur through PD-1/PD-L1 co-inhibitory binding and TCR activation.1 This system can be used to evaluate compounds to block PD-1/PD-L1 binding interactions or signaling. The Jurkat cells expressing PD-1 can be co-cultured with BW5147 TCS cells (SCC675) which will result in T-cell activation and downstream eGFP expression via NF-κB eGFP reporter construct, however, this result is inhibited by PD-1/PD-L1 binding interactions leading to a dampened eGFP response. Blocking PD-1/PD-L1 binding will result in improved eGFP expression. These unique interactions allow the co-culture system to be used for a diverse number of applications.

Source
Derived from BW5147 murine thymoma line. Cells were engineered to stably express an anti-human CD3 single chain fragment anchored to the cell membrane via a human CD14 stem. Cells were retrovirally transduced to express high levels of PD-L1.

References
Arifin MZ, Leitner J, Egan D, Waidhofer-Söllner P, Kolch W, Zhernovkov V, Steinberger P. 2024. BTLA and PD-1 signals attenuate TCR-mediated transcriptomic changes. iScience. 27(7):110253.

The BW5147 cells express PD-L1 and membrane bound anti-CD3 single chain antibody fragments fused to human CD14 in addition to the costimulatory ligand CD86 used to bind to TCR complexes and activate T-cells.

BW5147 Murine Thymoma T Cell Stimulator Expressing PD-L1 cells should be stored in liquid nitrogen. The cells can be cultured for at least 10 passages without significantly affecting cell marker expression and function.

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