Multiparameter flow cytometric analysis of CD4 and CD8 T cell subsets in young and old people

TLDR

T cell‑mediated immunity in the elderly is compromised, reflected in peripheral T cell composition, and polychromatic flow cytometry now enables detailed analysis of shifts among naïve, central memory, and effector memory subsets defined by CD45RA/CCR7 and additional costimulatory/coinhibitory markers. The study aims to build a differentiation scheme for CD4 and CD8 T cells by integrating multiple markers and functional assays, following the model of Romero et al. The authors employed polychromatic flow cytometry to analyze CD45RA/CCR7‑defined subsets, adding CD27, CD28, CD57, and KLRG‑1 markers, and incorporated proliferation and cytokine secretion assays to map differentiation pathways. They found that age‑related changes are driven by both subset distribution differences and inter‑subset differences, with CD8 cells showing more marked shifts, while CD4 cells exhibit more intra‑subset variation;.

Abstract

T cell-mediated immunity in elderly people is compromised in ways reflected in the composition of the peripheral T cell pool. The advent of polychromatic flow cytometry has made analysis of cell subsets feasible in unprecedented detail.Here we document shifts in subset distribution within naïve (N), central memory (CM) and effector memory (EM) cells defined by CD45RA and CCR7 expression in the elderly, additionally using the costimulatory receptors CD27 and CD28, as well as the coinhibitory receptors CD57 and KLRG-1, to further dissect these. Although differences between young and old were more marked in CD8 than in CD4 cells, a similar overall pattern prevailed in both. Thus, the use of all these markers together, and inclusion of assays of proliferation and cytokine secretion, may enable the construction of a differentiation scheme applicable to CD4 as well as CD8 cells, with the model (based on Romero et al.) suggesting the progression N-->CM-->EM1-->EM2-->pE1-->pE2-->EM4-->EM3-->E end-stage non-proliferative effector cells.Overall, the results suggest that both differences in subset distribution and differences between subsets are responsible for age-related changes in CD8 cells but that differences within rather than between subsets are more prominent for CD4 cells.