Natural killer cell receptor signaling and activation depend on cell cycle stages
10 affiliations
- 1Steve and Cindy Rasmussen Institute for Genomic Medicine, Abigail Wexner Research Institute at Nationwide Children's Hospital, Columbus, Ohio, United States of America.
- 2Biomedical Sciences Graduate Program, The Ohio State University, Columbus, Ohio, United States of America.
- 3Department of Microbiology and Immunology and the Parker Institute for Cancer Immunotherapy, University of California-San Francisco, San Francisco, California, United States of America.
- 4GIG Statistical Consulting, LLC, Columbus, Ohio, United States of America.
- 5Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan.
- 6Department of Pathology, Stanford University School of Medicine, Palo Alto, California, United States of America.
- 7Division of Hematology, The Ohio State University, Columbus, Ohio, United States of America.
- 8Pelotonia Institute for Immuno-Oncology, The James Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio, United States of America.
- 9Department of Pediatrics, College of Medicine, The Ohio State University, Columbus, Ohio, United States of America.
- 10Biophysics Program, The Ohio State University, Columbus, Ohio, United States of America.
Abstract
Natural Killer (NK) cells are lymphocytes of the innate immunity and provide key protection against infection and cancer. NK cells express a wide array of inhibitory and activating receptors on the cell surface including the activating receptor NKG2D. Receptor signaling in NK cells leads to rapid post-translational modification (e.g., phosphorylation) of sub-cellular signaling proteins within minutes of receptor stimulation that ultimately give rise to diverse effector functions including cell proliferation. Recent single-cell mass cytometry (i.e., CyTOF) experiments in macrophages showed variations of abundances of phosphorylated signaling proteins across cell cycle stages indicating a dependence of cell signaling kinetics on an order of magnitude slower kinetics (~ several hours) of cell cycle transitions. We investigated cell cycle dependence of NKG2D signaling kinetics in NK cells using longitudinal CyTOF measurements up to 256 minutes post stimulation performed on IL-2-treated NKG2D-stimulated primary human CD56dim NK cells. The CyTOF experiments revealed cell stage specific kinetics of signaling proteins such as pVav1, pErk, and pS6, with the degranulation marker CD107a showing its largest upregulation specifically in the combined G2 and M (G2 - M) stage. We developed stochastic minimal computational models coupling signaling and cell cycle processes to investigate the effect of cell cycle transitions on NK cell receptor induced signaling kinetics. These models suggest that the observed cell stage specific NK cell receptor signaling kinetics arise due to active modulation of the receptor signaling kinetics by cell cycle progression. The framework can be extended to study the interplay between receptor signaling and cell cycle progression in other cell types. Author summary: NK cells are innate lymphocytes that protect against infection and cancer. They express receptors such as NKG2D, which enable them to sense their environment and respond to healthy or diseased cells. Upon binding to cognate ligands, receptors such as NKG2D induce a series of rapid biochemical signaling reactions within minutes, regulating NK cell effector functions such as cell proliferation or cytokine release. In contrast, dividing NK cells progress through the cell cycle (G1 → S → G2 → M) on a much slower timescale of hours. Since protein synthesis, including that of signaling components, occurs during cell cycle progression, receptor signaling kinetics are likely to depend on cell cycle stage. We performed single-cell CyTOF experiments on IL-2-treated primary human NK cells stimulated with anti-NKG2D antibodies, measuring signaling and cell cycle-associated proteins over time. The results revealed cell-stage-specific signaling kinetics, including early activation and degranulation responses. We developed stochastic minimal models that couple receptor signaling with cell cycle transitions to quantify how cell cycle progression regulates signaling kinetics. Analysis of CyTOF data supports an active role of the cell cycle in modulating NKG2D signaling. This framework can be extended in other cell types.
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BibTeX
@article{wethington2026natural,
title = {{Natural killer cell receptor signaling and activation depend on cell cycle stages}},
author = {Darren Wethington and Indrani Nayak and Helle Jensen and William C Stewart and Oscar A Aguilar and Shih-Yu Chen and Garry P Nolan and Gregory K Behbehani and Lewis L Lanier and Jayajit Das},
journal = {PLOS Computational Biology},
year = {2026},
month = oct,
doi = {10.1371/journal.pcbi.1014825},
url = {https://journals.plos.org/ploscompbiol/article?id=10.1371/journal.pcbi.1014825},
note = {CC BY},
}
RIS
TY - JOUR
TI - Natural killer cell receptor signaling and activation depend on cell cycle stages
AU - Darren Wethington
AU - Indrani Nayak
AU - Helle Jensen
AU - William C Stewart
AU - Oscar A Aguilar
AU - Shih-Yu Chen
AU - Garry P Nolan
AU - Gregory K Behbehani
AU - Lewis L Lanier
AU - Jayajit Das
PY - 2026
DA - 2026/10/13
JO - PLOS Computational Biology
DO - 10.1371/journal.pcbi.1014825
UR - https://journals.plos.org/ploscompbiol/article?id=10.1371/journal.pcbi.1014825
AB - Natural Killer (NK) cells are lymphocytes of the innate immunity and provide key protection against infection and cancer. NK cells express a wide array of inhibitory and activating receptors on the cell surface including the activating receptor NKG2D. Receptor signaling in NK cells leads to rapid post-translational modification (e.g., phosphorylation) of sub-cellular signaling proteins within minutes of receptor stimulation that ultimately give rise to diverse effector functions including cell proliferation. Recent single-cell mass cytometry (i.e., CyTOF) experiments in macrophages showed variations of abundances of phosphorylated signaling proteins across cell cycle stages indicating a dependence of cell signaling kinetics on an order of magnitude slower kinetics (~ several hours) of cell cycle transitions. We investigated cell cycle dependence of NKG2D signaling kinetics in NK cells using longitudinal CyTOF measurements up to 256 minutes post stimulation performed on IL-2-treated NKG2D-stimulated primary human CD56dim NK cells. The CyTOF experiments revealed cell stage specific kinetics of signaling proteins such as pVav1, pErk, and pS6, with the degranulation marker CD107a showing its largest upregulation specifically in the combined G2 and M (G2 - M) stage. We developed stochastic minimal computational models coupling signaling and cell cycle processes to investigate the effect of cell cycle transitions on NK cell receptor induced signaling kinetics. These models suggest that the observed cell stage specific NK cell receptor signaling kinetics arise due to active modulation of the receptor signaling kinetics by cell cycle progression. The framework can be extended to study the interplay between receptor signaling and cell cycle progression in other cell types. Author summary: NK cells are innate lymphocytes that protect against infection and cancer. They express receptors such as NKG2D, which enable them to sense their environment and respond to healthy or diseased cells. Upon binding to cognate ligands, receptors such as NKG2D induce a series of rapid biochemical signaling reactions within minutes, regulating NK cell effector functions such as cell proliferation or cytokine release. In contrast, dividing NK cells progress through the cell cycle (G1 → S → G2 → M) on a much slower timescale of hours. Since protein synthesis, including that of signaling components, occurs during cell cycle progression, receptor signaling kinetics are likely to depend on cell cycle stage. We performed single-cell CyTOF experiments on IL-2-treated primary human NK cells stimulated with anti-NKG2D antibodies, measuring signaling and cell cycle-associated proteins over time. The results revealed cell-stage-specific signaling kinetics, including early activation and degranulation responses. We developed stochastic minimal models that couple receptor signaling with cell cycle transitions to quantify how cell cycle progression regulates signaling kinetics. Analysis of CyTOF data supports an active role of the cell cycle in modulating NKG2D signaling. This framework can be extended in other cell types.
SN - 1553-7358
SN - 1553-734X
ER -