Interleukin-10 (IL-10) is a pleiotropic cytokine which has potent inhibitory effects on macrophages and T cells, and contributes to the regulation of proliferation and differentiation of B cells. Resting HuT 78, a T-cell line derived from a Sezary lymphoma, produced significant amounts of IL-10 compared with another T-cell line, Jurkat. To elucidate the mechanisms by which the IL-10 is expressed, we have analyzed their activity in human T-cell lines. We report here evidence that members of the family of transcription factors nuclear factor-kappa B (NF-kappa B)/Rel can specifically recognize 3 identical sequences located in the 5'-regulatory region of IL-10 gene in resting HuT 78 cells, whereas Jurkat cells expressed high levels of NF-kappa B consisting of p65 and p50 only upon activation with tumor necrosis factor-alpha (TNF-alpha). In HuT 78 cells, p50 was the major component in the NF-kappa B complexes. Exogenous TNF-alpha and the monoclonal antibody to TNF-alpha did not affect IL-10 production and constitutive NF-kappa B binding levels in HuT 78 cells. This study is the first demonstration of a role for NF-kappa B in the IL-10 gene expression, and suggests that its expression does not require TNF-alpha. These novel findings may account for the specific IL-10 gene expression in T cells.