In view of the importance of vitamin A in the human immune system and the central role of interleukin-2 (IL-2) in the proliferation of T-lymphocytes, we examined the effects of all-trans-retinoic acid (ATRA) on the protein and gene expression of IL-2 in the human T-cell line HUT-78 when stimulated with either 12-O-tetradecanoylphorbol-13-acetate (TPA) or phytohemagglutinin (PHA). ATRA enhanced the production of IL-2 stimulated by TPA, but suppressed that stimulated by PHA. These findings were consistent with the results of a reverse transcription-polymerase chain reaction and quantitative real-time polymerase chain reaction examining IL-2 gene expression. ATRA augmented the gene expression of PKC-beta1 up-regulated by TPA and restored that suppressed by PHA but to below the control level. ATRA suppressed the c-fos gene expression up-regulated by PHA to a level of 36% of the control whereas it had no effect on the up-regulation by TPA. Since PKC- beta1 has been suggested to be important for the secretion and gene expression of IL-2 and since the activator protein-l binding site is present in the promoter of the IL-2 gene, these findings may explain the differences in ATRA's effects on TPA- and PHA-stimulated IL-2 expression. These results suggest that ATRA affects the production of IL-2 by T-lymphocytes in a stimulus-dependent manner.