Multiple mechanisms regulate the nanoscale organization of PD-L1 at the cell surface
- 1Department of Medical BioSciences, Radboud Institute for Medical Innovation, Radboud University Medical Center, Nijmegen, The Netherlands
Abstract
The immune checkpoint protein PD-L1 plays a pivotal role in tumor immune evasion by binding to PD-1 on immune cells, including T lymphocytes. While the expression and function of PD-L1 have been well studied, the importance of its spatial organization on the cell surface of tumor cells remains poorly understood. In this study, we used super-resolution microscopy combined with biochemical perturbations to investigate the factors regulating PD-L1 clustering and its effects on PD-1 binding and T cell inhibition. We found that PD-L1 is organized into nanoscale clusters at the plasma membrane, with distinct regulatory roles for the actin cytoskeleton, galectin-3, and cholesterol. Disruption of cortical actin increased PD-L1 cluster size, while galectin-3 promoted smaller, denser clusters and increased PD-L1 lateral mobility. Cholesterol depletion reduced PD-L1 cluster size and number and impaired PD-1 binding. These findings indicate that PD-L1 surface organization is collectively regulated by the actin cytoskeleton, galectin-3, and membrane cholesterol within the plasma membrane of tumour cells. Our results provide new insights into the dynamic regulation of PD-L1 and its potential as a therapeutic target in cancer immunotherapy.
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BibTeX
@misc{franken2026multiple,
title = {{Multiple mechanisms regulate the nanoscale organization of PD-L1 at the cell surface}},
author = {G. A. Franken and A. B. Arp and D. Cerina and V. M. R. van Esch and B. Scheijen and A. B. van Spriel},
howpublished = {bioRxiv},
year = {2026},
month = aug,
doi = {10.64898/2026.08.31.748200},
url = {https://www.biorxiv.org/content/10.64898/2026.08.31.748200v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Multiple mechanisms regulate the nanoscale organization of PD-L1 at the cell surface
AU - G. A. Franken
AU - A. B. Arp
AU - D. Cerina
AU - V. M. R. van Esch
AU - B. Scheijen
AU - A. B. van Spriel
PY - 2026
DA - 2026/08/31
PB - bioRxiv
DO - 10.64898/2026.08.31.748200
UR - https://www.biorxiv.org/content/10.64898/2026.08.31.748200v1
AB - The immune checkpoint protein PD-L1 plays a pivotal role in tumor immune evasion by binding to PD-1 on immune cells, including T lymphocytes. While the expression and function of PD-L1 have been well studied, the importance of its spatial organization on the cell surface of tumor cells remains poorly understood. In this study, we used super-resolution microscopy combined with biochemical perturbations to investigate the factors regulating PD-L1 clustering and its effects on PD-1 binding and T cell inhibition. We found that PD-L1 is organized into nanoscale clusters at the plasma membrane, with distinct regulatory roles for the actin cytoskeleton, galectin-3, and cholesterol. Disruption of cortical actin increased PD-L1 cluster size, while galectin-3 promoted smaller, denser clusters and increased PD-L1 lateral mobility. Cholesterol depletion reduced PD-L1 cluster size and number and impaired PD-1 binding. These findings indicate that PD-L1 surface organization is collectively regulated by the actin cytoskeleton, galectin-3, and membrane cholesterol within the plasma membrane of tumour cells. Our results provide new insights into the dynamic regulation of PD-L1 and its potential as a therapeutic target in cancer immunotherapy.
ER -