Aging-associated subpopulations of human CD8+ T-lymphocytes identified by their CD28 and CD57 phenotypes

doi:10.1016/j.archger.2015.08.007

Archives of Gerontology and Geriatrics

Volume 61, Issue 3, November–December 2015, Pages 494-502

https://doi.org/10.1016/j.archger.2015.08.007 Get rights and content

Highlights

•
Among the CD8 T-lymphocytes, only CD57+ subpopulations increase with aging.
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Characteristics of replicative cell aging are most pronounced in CD28+CD57+ cells.
•
Our findings favor the presence of senescent cells in vivo.

Abstract

Background

During organismal aging, human T-cells shift towards less functional phenotypes, often called senescent cells. As these cells have not been well characterized, we aimed to relate surface markers of human T-cell senescence with characteristics of in vitro cellular aging and to further characterize these cells.

Methods

We identified, by flow cytometry, subpopulations of CD8+ T-cells based on CD57 and CD28 expression, and tested them for some markers of cellular senescence, apoptosis, differentiation and homing.

Results

Elderly persons presented significantly higher proportions not only of CD28−CD57+, but also of CD28+CD57+ cells. CD28+CD57+ cells had the highest expression of p16, p21, Bcl-2, CD95, CD45RO, CCR5 and PD-1, thereby arguing in favor of a senescent phenotype.

Conclusion

Among CD8+ T-lymphocytes, CD28+CD57+ cells represent a subset with some senescent features that are distinct from the CD28−CD57+ cells.

Introduction

Reports on senescence of human cells have highly relied on in vitro observations, which might, however, not be true reflections of in vivo characteristics (Rodier and Campisi, 2011). As a consequence, the contribution of senescent cells to the human aging process has remained controversial. On the other hand, organismal aging invariably leads to a decrease in functionality. In the immune system, this results in immunosenescence, a complex process that includes a shift towards less functional T-cells that are often designated as senescent. Cell surface markers, mainly absence of CD28 or expression of CD57, have been used to identify T lymphocytes as senescent in vitro (Brenchley et al., 2003, Effros, 1997).

Working with CD3+ T-cells, we have found the combined expression of CD28 and CD57 useful in distinguishing senescent subpopulations. In elderly compared with young persons, we have observed not only a higher prevalence of CD28−CD57+ cells, as expected, but also of a small subpopulation that is CD28+CD57+. (Onyema et al., 2012). The attention on CD28+CD57+ cells was first drawn in 2003 by Brenchley et al., who observed in six healthy individuals that 8% of memory CD8+ T cells consisted of the CD28+CD57+ phenotype. In their study the expression of CD57 on T-cells was related to short telomeres and loss of proliferation capacity, possibly linking its presence to senescence (Brenchley et al., 2003). CD28+CD57+ T-cells differ from the more prevalent CD28−CD57+ cells, which are known to correspond to a highly differentiated subpopulation with effector function (Onyema et al., 2012). While CD28−CD57+ cells have been related to persistent viral infections and an immune risk profile (IRP) (Olsson et al., 2000, Wikby et al., 2002), we have found that CMV seropositivity was not related to the prevalence of CD28+CD57+ cells (Onyema et al., 2012). Based on these observations, CD28+CD57+ cells might be a different phenotype of senescent T-cells. However, their nature remains to be further elaborated.

Here, focusing on CD8+ T lymphocytes, we looked at various characteristics related to cellular senescence, apoptosis, T-cell differentiation and homing. We observed that the CD28+CD57+ subpopulation increased with aging, presented some features of a senescent cell phenotype, and has differentiation characteristics of memory T-cells destined for tissue migration.

Section snippets

Participants

One group of young (N = 11) and two groups of elderly participants (N = 11 + 11) were recruited. First, to study the protein expression pattern of senescence and apoptosis modulators in CD8+ subpopulations of T-cells we recruited 11 young (median age 24 ± 2.3 years; 5 females and 6 males) and 11 elderly (E1) (median age 84 ± 1.8 years; 5 females and 6 males) Caucasians from the Belgian population. These 22 participants were the same as in our previous study; earlier studies have shown that the sample

Results

The gating procedure for the delineation of the different subpopulations is shown in Fig. 1A. Fig. 1B shows the proportion of CD8+ T-cells according to their combined expression of the surface markers CD28 and CD57 in young (N = 11) and E1 old (N = 11) participants. Cells expressing CD57 were more frequent in the older participants, irrespective of their CD28 status, while CD28− cells were more frequent only if they also expressed CD57. The frequency of the various CD8+ subpopulations in the second

Discussion

When considering the cell surface markers CD28 and CD57, we observed in CD8+ T-cells from old (E1) compared to young subjects not only significantly higher proportions of the CD28−CD57+ cell population (34.5% versus 6.0%), as expected, but also of a small population that was characterized as CD28+CD57+ (3.0% versus 0.8%). Both subpopulations were CD57+, emphasizing the importance of CD57 expression in T-lymphocyte aging, independent of the CD28 phenotype.

At the protein level, the highest

Conflict of interest

None of the authors has a conflict of interest to report.

Acknowledgments

We acknowledge Angelo Willems, Wim Renmans and Carlo Heirman, for their expert assistance with cell sorting, flow cytometry and PCR respectively. The study was supported by a scientific grant from the “Wetenschappelijk Fonds Willy Gepts” from the Universitair Ziekenhuis Brussel.

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Aging-associated subpopulations of human CD8+ T-lymphocytes identified by their CD28 and CD57 phenotypes

Highlights

Abstract

Background

Methods

Results

Conclusion

Introduction

Section snippets

Participants

Results

Discussion

Conflict of interest

Acknowledgments

Cell

Blood

Developmental and Comparative Immunology

American Journal of Human Genetics

Blood

Molecular Cell

Cell

Biochemical and Biophysical Research Communications

Journal of Biological Chemistry

Human Immunology

Brain, Behavior, & Immunity

Experimental Gerontology

Blood

Experimental Gerontology

Increased apoptosis of T cell subsets in aging humans: altered expression of Fas (CD95), Fas ligand, Bcl-2, and Bax

The Journal of Immunology

Loss of CD28 expression on CD8(+) T cells is induced by IL-2 receptor gamma chain signalling cytokines and type I IFN, and increases susceptibility to activation-induced apoptosis

International Immunology

CD95 promotes tumour growth

Nature

Proliferation and interleukin 5 production by CD8hi CD57+ T cells

European Journal of Immunology

PD-1 expression on HIV-specific T cells is associated with T-cell exhaustion and disease progression

Nature

A biomarker that identifies senescent human cells in culture and in aging skin in vivo

Proceedings of the National Academy of Sciences of the United States of America

Phosphorylation of p53 at serine 37 is important for transcriptional activity and regulation in response to DNA damage

Oncogene

Involvement of the Ink4 proteins p16 and p15 in T-lymphocyte senescence

Oncogene

Functional expression of the chemokine receptor CCR5 on virus epitope-specific memory and effector CD8+ T cells

The Journal of Immunology