CD4+ T cell surface alpha enolase is lower in older adults

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 <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 alpha-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 alpha-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 et-enolase expression was observed in age-matched patients with cardiovascular disease (p < 0.05). An immune-modulatory role has been proposed for surface alpha-enolase and our findings of decreased expression suggest that deficits in surface alpha-enolase merit investigation in the context of immune dysfunction during ageing and vascular disease. (C) 2015 Elsevier Ireland Ltd. All rights reserved.