Over the years of testing biocompatibility of endodontic filling materials, little attention has been paid to the potential adverse influences on the function of the immune system. Therefore, the purpose of this study was to investigate the extent to which extractable components of some commonly used root canal sealing materials (ERCS) may interfere with immunocompetent cells in vitro. The potential of these materials to cause delayed-type hypersensitivity (DTH) was also addressed in a rat model system. Extractable components were drawn in cell culture medium from freshly mixed or set material of AH 26. Grossman's sealer, Endomethasone, and Apexit. In-vitro assays included either spleen cells or rat pulp tissue cells that were released following enzymatic digestion with collagenase. Purified T cells for the pulpal cell assay were obtained from rat mesenteric lymph nodes. The effect of ERCS on the proliferation of concanavalin A (con A) stimulated spleen cell was measured by 3H-thymidine incorporation. Pulpal accessory cell function was monitored by the capacity of pulpal cells, pretreated with components of ERCS, to provide signals to con A stimulated T cells. DTH was tested after subcutaneous implantation of root canal sealers (RCS) in rats and challenge by ear injection. Pretreatment of pulpal cells with low dilutions of eluates from extracted AH 26 and Endomethasone resulted in a strong reduction of the T cell proliferation rate. The effect was considerably reduced (P < 0.01) when extracts of the solid material were employed. Extracts of Grossmans' sealer and Apexit affected T cell proliferation only to a limited extent in the pulpal cell assay. In general, assays on spleen cells showed a similar profile, although increased cell division was induced by Grossman's sealer at high eluate dilutions and a concentration-dependent decrease of cell division at lower concentrations of this material. ERCS evoked both immunosuppression and, in some instances, immunostimulation, but they did not release DTH.