This study used a panel of mAb and multiparameter flow cytometry to assess the composition of PBL from healthy aged individuals. The results showed that while total lymphocyte numbers altered only marginally in the aged (> or = 70 years) there were significant changes in the distribution of various sub-populations; for example, there were lower numbers of CD3+ and CD8+ cells, and higher numbers of CD16+ (NK) cells. As a direct result of these changes the numbers of CD2+ cells remained unchanged in the aged compared with young adult controls (18-25 years). The number of CD4+ T cells expressing CD45RO isoform (memory cells) exceeded the number of CD4+CD45RA+ (naive) cells in aged donors, whereas the converse was true for young donors. In addition, associated with this increase in CD45RO+ cells, the aged showed an expanded population of CD45RO+ T cells displaying low surface levels of CD45RO. These data suggest that shifts in the distribution of regulatory T cell subsets may play a role in age-related changes in immune response.