SA-β-Gal stratification reveals senescence-associated heterogeneity and differential cell death susceptibility in human CD8 effector T cells
- 1Rutgers School of Graduate Studies, Rutgers Biomedical and Health Sciences, Rutgers University
- 2Department of Pathology, Immunology, and Laboratory Medicine, Rutgers-New Jersey Medical School
- 3Department of Microbiology, Biochemistry and Molecular Genetics, Center for Cell Signaling, Rutgers-New Jersey Medical School
Abstract
Introduction: Cellular senescence in human T cells remains incompletely defined, particularly with respect to how senescence-associated states influence susceptibility to cell death.
Methods: We used flow-cytometric SPiDER-based detection of senescence-associated β-galactosidase (SA-β-Gal) activity to profile human CD8+ T-cell subsets from young and old donors. We stratified effector memory (CD8 EM ) and effector memory re-expressing CD45RA (CD8 EMRA ) subsets by the lowest (bottom 25%) and highest (top 25%) quartiles of SA-β-Gal signal.
Results: CD8 EM and CD8 EMRA cells displayed the greatest within-subset heterogeneity in SA-β-Gal activity. The SA-β-Gal low and high populations occupied distinct regions in UMAP projections and were reproducibly enriched in different FlowSOM metaclusters, indicating that SA-β-Gal activity captures structured phenotypic states within differentiated CD8+ T-cell compartments. These states differed in senescence-associated markers, including CD27, CD28, CD57, and CX3CR1, as well as granzyme-associated programs involving Gzm A, B and K, whereas chronological age had only a limited effect once subsets were analyzed separately. Functionally, nutrient-limited conditions unmasked marked differences in cell death susceptibility between these populations. Following PMA/ionomycin stimulation, SA-β-Gal high CD8 EM and CD8 EMRA cells underwent accelerated stimulation-induced cell death, showed increased caspase-3/7 activation, and exhibited lower post-treatment γH2AX signal relative to SA-β-Gal low counterparts. In contrast, in the absence of stimulation-induced energetic demand, SA-β-Gal high cells displayed greater resistance to cell death.
Discussion: Together, SA-β-Gal stratification resolves structured heterogeneity within human CD8 effector T cells and supports a context-dependent model in which senescence-associated effector states become selectively vulnerable to stimulation-induced death under metabolic stress.
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BibTeX
@article{garzamartinez2026saβgal,
title = {{SA-β-Gal stratification reveals senescence-associated heterogeneity and differential cell death susceptibility in human CD8 effector T cells}},
author = {Luis Garza-Martínez and Mark Apadeer and Nica Mariz Aquino and Paolo S. Turano and Elizabeth Akbulut and Dillon Oswald and Sukhwinder Singh and Patricia Fitzgerald-Bocarsly},
journal = {Frontiers in Immunology},
year = {2026},
month = oct,
doi = {10.3389/fimmu.2026.1985303},
url = {https://www.frontiersin.org/articles/10.3389/fimmu.2026.1985303/full},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - SA-β-Gal stratification reveals senescence-associated heterogeneity and differential cell death susceptibility in human CD8 effector T cells
AU - Luis Garza-Martínez
AU - Mark Apadeer
AU - Nica Mariz Aquino
AU - Paolo S. Turano
AU - Elizabeth Akbulut
AU - Dillon Oswald
AU - Sukhwinder Singh
AU - Patricia Fitzgerald-Bocarsly
PY - 2026
DA - 2026/10/26
JO - Frontiers in Immunology
DO - 10.3389/fimmu.2026.1985303
UR - https://www.frontiersin.org/articles/10.3389/fimmu.2026.1985303/full
AB - Introduction: Cellular senescence in human T cells remains incompletely defined, particularly with respect to how senescence-associated states influence susceptibility to cell death. Methods: We used flow-cytometric SPiDER-based detection of senescence-associated β-galactosidase (SA-β-Gal) activity to profile human CD8+ T-cell subsets from young and old donors. We stratified effector memory (CD8 EM ) and effector memory re-expressing CD45RA (CD8 EMRA ) subsets by the lowest (bottom 25%) and highest (top 25%) quartiles of SA-β-Gal signal. Results: CD8 EM and CD8 EMRA cells displayed the greatest within-subset heterogeneity in SA-β-Gal activity. The SA-β-Gal low and high populations occupied distinct regions in UMAP projections and were reproducibly enriched in different FlowSOM metaclusters, indicating that SA-β-Gal activity captures structured phenotypic states within differentiated CD8+ T-cell compartments. These states differed in senescence-associated markers, including CD27, CD28, CD57, and CX3CR1, as well as granzyme-associated programs involving Gzm A, B and K, whereas chronological age had only a limited effect once subsets were analyzed separately. Functionally, nutrient-limited conditions unmasked marked differences in cell death susceptibility between these populations. Following PMA/ionomycin stimulation, SA-β-Gal high CD8 EM and CD8 EMRA cells underwent accelerated stimulation-induced cell death, showed increased caspase-3/7 activation, and exhibited lower post-treatment γH2AX signal relative to SA-β-Gal low counterparts. In contrast, in the absence of stimulation-induced energetic demand, SA-β-Gal high cells displayed greater resistance to cell death. Discussion: Together, SA-β-Gal stratification resolves structured heterogeneity within human CD8 effector T cells and supports a context-dependent model in which senescence-associated effector states become selectively vulnerable to stimulation-induced death under metabolic stress.
SN - 1664-3224
ER -