A comparative study of the JAM test and 51Cr-release assay to assess the cytotoxicity of dendritic cells on hematopoietic tumor cells

Dendritic cells (DCs) are potent antigen presenting cells and possess a direct anti-tumor cytotoxic ability. Nevertheless, the mechanism of Anti-tumor cytotoxicity by DCs and the methods for its evaluation are not fully elucidated. In order to clarify this mechanism of cytotoxicity, we examined the ability of DCs 1) to suppress [H-3] thymidine (H-3-TdR) uptake by tumor cells; 2) to induce cytolysis on Cr-51-labeled tumor cells; 3) and to induce DNA fragmentation on H-3-TdR labeled tumor cells (JAM test). Cytolysis and DNA fragmentation are markers of necrotic and apoptotic mechanisms of cytotoxicity in vitro, respectively. DCs inhibited approximately 38.6% to 54.8% of the growth of B4D6, NB4, U937, and Daudi cells as evaluated by the uptake of H-3-TdR. However no cytolysis was verified by Cr-51-release assay. On the other hand, cytotoxicity rates found using the JAM test ranged from 3 to 81% depending on the cell line and the effector to target cell ratio. The discrepancy of cytotoxicity between Cr-51-release assay and the JAM test may be due to the phagocytosis of apoptotic tumor cells or the absorption of released Cr-51 by DCs surrounding the target cells. In conclusion, the JAM test was more sensitive than the 4-h and the 10-h Cr-51-release assay to investigate cytotoxicity mediated by DCs toward hematopoietic tumor cell lines in vitro.