Abstract
To identify novel cell ageing markers in order to gain insight into ageing mechanisms, we adopted membrane enrichment and comparison of the CD4+ T cell membrane proteome (purified by cell surface labelling using Sulfo-NHS-SS-Biotin reagent) between healthy young (n = 9, 20–25 years) and older (n = 10; 50–70 years) male adults. Following two-dimensional gel electrophoresis (2DE) to separate pooled membrane proteins in triplicates, the identity of protein spots with age-dependent differences (p < 0.05 and >1.4 fold difference) was determined using liquid chromatography–mass spectrometry (LC–MS/MS). Seventeen protein spot density differences (ten increased and seven decreased in the older adult group) were observed between young and older adults. From spot intensity analysis, CD4+ T cell surface α-enolase was decreased in expression by 1.5 fold in the older age group; this was verified by flow cytometry (n = 22) and qPCR with significantly lower expression of cellular α-enolase mRNA and protein compared to young adult CD4+ T cells (p < 0.05). In an independent age-matched case-control study, lower CD4+ T cell surface α-enolase expression was observed in age-matched patients with cardiovascular disease (p < 0.05). An immune-modulatory role has been proposed for surface α-enolase and our findings of decreased expression suggest that deficits in surface α-enolase merit investigation in the context of immune dysfunction during ageing and vascular disease.
Original language | English |
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Pages (from-to) | 56-62 |
Journal | Mechanisms of Ageing and Development |
Volume | 152 |
Early online date | 1 Oct 2015 |
DOIs | |
Publication status | Published - Dec 2015 |
Keywords
- Enolase
- CD4+T cell
- Plasma membrane proteomics
- 2D gel electrophoresis
- Phenotype