To study the decreasing responsiveness of the immune system during aging, influenza virus specific cellular immunity was investigated in a cohort of healthy blood donors between 18 and 70 years of age. The percentage of influenza A virus specific T cells was determined by flow cytometry and found not to change during aging. After stimulation with phorbol 12-myristate 13-acetate and ionomycin, an increase in the percentage of IFN-gamma and IL-4 producing CD8(+) T cells was observed during aging. In addition, the cytotoxic T lymphocyte (CTL) activity was investigated in two additional groups of five donors, 18-20 and 68-70 years of age. The lytic capacity of purified CD8(+) T cells, after in vitro stimulation of peripheral blood mononuclear cells with influenza A virus, seemed lower in 68- to 70-year-old donors than in 18- to 20-year-old donors. Therefore we conclude that the reduced CTL activity in the elderly is not the result of a lower frequency of virus-specific T cells, but more likely the result of impaired antigen-specific proliferation or lower lytic capacity of these cells.