Purpose: To investigate the ability of human lymphocytes labeled with DNA-incorporated 125I to exert an inhibitory (antiproliferative) bystander effect on co-cultured human colon adenocarcinoma LS174T cells in vitro.Materials and methods: Human peripheral blood lymphocytes were stimulated to synthesize DNA in the presence of phytohemagglutinin (PHA) and labeled with 5-[125I]iodo-2′-deoxyuridine. Human colon adenocarcinoma LS174T cells were co-cultured with the 125I-labeled lymphocytes in various ratios for 5 days and the proliferation of the LS174T cells was assessed. Further, the supernatant media from these co-cultures were: (i) Transferred to LS174T cells and their proliferation measured after 5 days, (ii) used to assess the clonogenic survival of LS174T cells, and (iii) screened for factors that suppress growth.Results: A significant reduction in the proliferation of LS174T cells was observed when co-cultured either with 125I-labeled lymphocytes (56 ± 3.5%) or the supernatant media (52.5 ± 1.3%) obtained from these co-cultures. Clonogenic survival of LS174T cells grown in the supernatant media corroborated the decrease in tumor cell growth.Conclusion: The observed reduction in the proliferation of LS174T cells in presence of 125I-labeled lymphocytes or media obtained from such co-cultures can be attributed to an inhibitory (antiproliferative) bystander effect, probably mediated by factor(s) released from the dying 125I-labeled lymphocytes.