A novel method for autophagy detection in primary cells: Impaired levels of macroautophagy in immunosenescent T cells
Open accessAbstract
Autophagy is a conserved constitutive cellular process, responsible for the degradation of dysfunctional proteins and organelles. Autophagy plays a role in many diseases such as neurodegeneration and cancer; however, to date, conventional autophagy detection techniques are not suitable for clinical samples. We have developed a high throughput, statistically robust technique that quantitates autophagy in primary human leukocytes using the Image stream, an imaging flow cytometer. We validate this method on cell lines and primary cells knocked down for essential autophagy genes. Also, using this method we show that T cells have higher autophagic activity than B cells. Furthermore our results indicate that healthy primary senescent CD8(+) T cells have decreased autophagic levels correlating with increased DNA damage, which may explain features of the senescent immune system and its declining function with age. This technique will allow us, for the first time, to measure autophagy levels in diseases with a known link to autophagy, while also determining the contribution of autophagy to the efficacy of drugs.
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Studies of human T cells have associated aging with changes in autophagy, membrane raft properties, and reactive oxygen production, but these examined total T cell populations irrespective of naive or memory phenotype and are difficult to interpret.
“Studies of human T cells have associated aging with changes in autophagy, in membrane properties to form rafts, and in reactive oxygen production (Fulop et al., 2014; Phadwal et al., 2012); however, these studies examined total T cell populations, irrespective of naive or memory phenotypes, and are therefore difficult to interpret.”
From Successful and Maladaptive T Cell Aging (2017) · cites this paper as Phadwal et al., 2012
Senescent CD8+ T cells display both low autophagic activity and lack mTORC1 activity.
“However senescent CD8 + T cells displayed both low autophagic activity [25, 65] and also lack mTORC1 activity [25].”
From Senescence of T Lymphocytes: Implications for Enhancing Human Immunity (2016) · cites this paper as [65]
Autophagy falls with age, including in human CD8+ T cells.
“Autophagy, degrading bulk cytoplasmic material by delivering it to the lysosomes, falls with age, including in human CD8+ T cells [105].”
From Evolution of the immune system in humans from infancy to old age (2015) · cites this paper as [105]
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References 40 · Cited by 155
- Authors
- Phadwal, K., Alegre-Abarrategui, J., Watson, A. S., Pike, L., Anbalagan, S., Hammond, E. M., Wade-Martins, R., McMichael, A., Klenerman, P., & Simon, A. K.
- Affiliations
- John Radcliffe Hospital (GB)
- University of Oxford (GB)
- Corresponding author
- Simon, Anna Katharina
- ORCID
- Venue
- Autophagy
- Publisher
- Informa UK Limited
- Published
- 2012-04
- Volume
- 8
- Issue
- 4
- Pages
- 677-689
- ISSN
- 1554-8627 (ISSN-L)
- 1554-8635
- PMC
- PMC3405842
- OpenAlex
- W2037959286
- Cited by
- 155
- References
- 40
- Citations by year
- 2026: 5 · 2025: 5 · 2024: 6 · 2023: 14 · 2022: 8 · 2021: 6 · 2020: 13 · 2019: 16 · 2018: 11 · 2017: 14 · 2016: 13 · 2015: 14 · 2014: 20 · 2013: 5 · 2012: 5
- FWCI
- 10.45
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- cc-by-nc
- Locations
- Autophagy (publishedVersion, cc-by-nc)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- MDC Repository (Max-Delbrueck-Center for Molecular Medicine) (acceptedVersion)
- Spiral (Imperial College London) (submittedVersion, cc-by-nc)
- Funders
- Wellcome Trust (091663)
- National Institute for Health and Care Research (NF-SI-0510-10204)
- Parkinson's UK (J-0901)
- Natural Sciences and Engineering Research Council of Canada
- Biotechnology and Biological Sciences Research Council (BB/G021422/1)
- Topics
- Health Sciences › Medicine › Epidemiology › Autophagy in Disease and Therapy
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Cell Image Analysis Techniques
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Aging › Genetics, Aging, and Longevity in Model Organisms
- Keywords
- imaging flow cytometry, high-throughput quantification, T cells, B cells, CD8+ T cell senescence, DNA damage, aging immune system, drug efficacy
- MeSH terms
- Adult, Animals, Autophagy, B-Lymphocytes, Biological Assay, Cells, Cultured, Histones, Humans, Middle Aged, T-Lymphocytes, Reproducibility of Results, Cellular Senescence, Mice, HEK293 Cells
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