We describe here a novel myelomonocytic cell line (OTT1) obtained from primary cultures of mouse bone marrow cells infected with a retroviral vector carrying the mouse interleukin (IL)-1 alpha gene. OTT1 cells are dependent for their survival and proliferation on IL-3, granulocyte-macrophage colony-stimulating factor (GM-CSF) or, unexpectedly, IL-5. Despite their IL-5 dependency, OTT1 cells form colonies showing predominantly monocyte maturation when plated in methylcellulose. It is suggested that constitutive expression of the exogenous IL-1 alpha gene may predispose to a monocytic phenotype. OTT1 cells should be a useful experimental model to investigate the molecular mechanisms of IL-5 signal transduction and the possible interrelationships between this signal pathway and those utilized by IL-3 and GM-CSF.