A cytokine-producing profile of anti-type 2 t cells induced in normal mice by treatment with benzoylmesaconine (BEN)

Marked protective effects of CD4+ anti-type 2 Tcells (BEN-Tcells), which were generated in spleens of mice treated with BEN (a non-toxic alkarold extracted from Aconiti tuber), have been shown in thermally injured mice infected with herpes simplex virus type 1. In the present study BEN-Tcells were characterized by their cytokine-producing profile. Purified BEN-Tcells were prepared from mice treated with BEN (Tsumura Co., Tokyo, Japan), as described. The anti-type 2 T cell activity was analyzed by a modified mixed lymphocyte reaction supplemented with T6S cells, a clone of burn-associated CDStype 2 Tcells. Activities of various cytokines were assayed as described previously. Cytokine mRNA expression of BEN-T cells was determined by RT-PCR method. As controls, LB2 cells, obtained from ATCC, and T6S cells were used as authentic Th1 and Th2 cells, respectively. BEN-Tcells (1x106 cells/ml) produced 170 U/ml of IFN-y when they were cultured with anti-CD3 mAb(2.5 ng/ml) for 48 hrs. However, the activities of IL-2. IL-4 and IL-10 were not detected in their culture fluids. IL-2 (160 U/ml) and IFN-y (190 U/ml) were produced by Th1 cells (1x106 cells/ml) stimulated the mAb. Th2 cells (1x106 cells/ml) produced IL4 (80 U/ml) and IL-10 (70 U/ml), but not produce IFN-r and IL-2, after the stimulation. BEN-T cells expressed mRNA for only IFN-r. when Th1 cells expressed for IFN-y mRNA and IL-2 mRNA and Th2 cells expressed for IL-4 mRNA and IL-10 mRNA. These results may suggest that BEN-T cells are a new CD4T cell subset, not belonging to Th1 and Th2 cells.