We have shown that tumor necrosis factor alpha (TNF alpha) and interleukin 2 (IL-2) are complementary for stimulation of growth of the hematopoietic stem cell line, LyD9. Neither TNF alpha nor IL-2 alone could stimulate the proliferation of LyD9 cells even after pre-incubation with these growth factors. The number of high-affinity IL-2 receptors on LyD9 cells did not increase after incubation with IL-2 and TNF alpha. These results suggest that the proliferative response of LyD9 by TNF alpha and IL-2 is not mediated by receptor inducing activities. We used the induction of the proto-oncogenes c-myc and c-pim to characterize the proliferative stimulation by IL-2 and TNF alpha. Northern blot analysis revealed that the simultaneous addition of IL-2 and TNF alpha was more efficient than IL-2 alone for c-myc mRNA induction. However, the addition of TNF alpha and IL-2 could not increase c-pim mRNA more than the level induced by IL-2 alone. The results indicate that the two growth factors complement each other by transducing different types of growth signal.