The present experiments were designed to perform a further investigation of the cell lineage of lymphokine-activated killer (LAK) cells. In the presence of adherent cells both T and not-T cells, separated on the basis of rosette formation with sheep erythrocytes (E rosettes), generated LAK activity after short-term culture in recombinant interleukin-2 in 5 different individuals tested. Since at the termination of the culture more than 98% of cells were T11-positive, it is evident that both LAK precursor and effector cells may belong to the T cell lineage. By applying a culture technique which allows the clonal expansion of virtually all T cells, we further selected and analyzed T cell clones with LAK activity. Under the culture conditions used, LAK clones represented approximately 4% of all proliferating clones. All had cytolytic activity against K562 target cells as well and also released large amounts of gamma-interferon following phytohemagglutinin stimulation.