期刊
AUTOPHAGY
卷 8, 期 4, 页码 677-689出版社
LANDES BIOSCIENCE
DOI: 10.4161/auto.18935
关键词
autophagy detection; ImageStream; PBMCs; T cells; immunosenescence
类别
资金
- Biomedical Research Centre Oxford (NIHR)
- Wellcome Trust
- NSERC
- BBSRC
- Biotechnology and Biological Sciences Research Council [BB/G021422/1] Funding Source: researchfish
- Medical Research Council [G1000800a] Funding Source: researchfish
- National Institute for Health Research [NF-SI-0510-10204] Funding Source: researchfish
- Parkinson's UK [J-0901] Funding Source: researchfish
- BBSRC [BB/G021422/1] Funding Source: UKRI
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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