The proliferative kinetics of the leukemic cells in the cerebrospinal fluid (CSF) were studied in three adults with CNS leukemia after 18 and 8 months while receiving an intensive treatment regimen. One (J.L.) had acute myeloblastic leukemia and developed CNS disease after 5 years' treatment with arabinosylcytosine and 6-thioguanine. All were in marrow remission at the time of study. G.H. had had no therapy for CNS disease, A.K. had had intrathecal methotrexate 2 months previously, and J.L. had recently completed radiotherapy (1900 R) to the head. An Ommaya reservoir was placed in a lateral ventricle and 3H-thymidine was injected into the resrvoir every 12 hours for 10 days; samples of cells were obtained by lumbar puncture periodically for autoradiography. In all patients the flash 3H-thymidine labeling index (LI) of the leukemic cells was less than 2% (determined in vitro) and the mitotic index less than 0.1%. After 10 days of 3H-thymidine injections in vivo, the LIs of the leukemic cells were 55%, 36%, and 21% in G.H., A.K., and J.L., respectively. These findings indicate that leukemic cells may proliferate very slowly in the CNS, and stress the difficulty of eradicating CNS leukemia with chemotherapeutic agents which are only active against proliferating cells.