The molecular mechanism responsible for the age-dependent decline of natural killer (NK) cell response was examined. The responses of NK cells to poly(I:C), Con A, and LPS were compared between the young and the aged animals. We observed that both basal and induced NK cell activity declined with age. To further determine the basis for this change, an effort was made to identify the cytokine genes that might be involved. In addition, the capacity of acetyl-L-carnitine (ALC) to restore NK cell activity was evaluated. In this study, 51Cr release assay and reverse transcriptase-polymerase chain reaction assay were used to measure the NK cell activity and the changes in cytokine gene expression, respectively. Our results show that the decline in NK cell response to poly(I:C) and LPS in aged animals was correlated with decreased expression of the IFNγ gene. The decline in the NK cell response to Con A in aged animals was, however, independent of the expression of IL-2 or IFN-γ genes. Although ALC was reported to restore functions of T cells and macrophages in aged animals, we found that long-term treatment (4 months) with ALC had no effect on either the basal or the induced age-dependent loss of NK cell activity. Our observations suggest that cytokine gene therapy could be a potential approach to improving or even preventing the decline of certain immune functions in the elderly.