A transfer model for studying both the development and prevention of diabetes in rats is described in detail. Diabetes was induced in BBDR rats by combining RT6-depletion with PolyI:C treatment. Autoreactive cells were isolated from acutely diabetic donors, reactivated in vitro and transferred intravenously into young (<34-day-old) BBDP rats. Accelerated diabetes occurred 13+/-3 days or 18+/-4 days after transfer of reactivated splenocytes or purified T cells (42/43 or 26/27 recipients, respectively). Freshly isolated mesenteric and splenic leukocytes from adult, healthy BBDR rats prevented spontaneous diabetes in BBDP rats, but were not able to prevent the accelerated diabetes when co-transferred with the autoreactive cells. By contrast, diabetes was significantly delayed (P<0.001) when protective cells were transferred 4 days prior to the autoreactive cells (16+/-3 days). In vivo tracking studies of the two types of transferred cells suggest different homing patterns which may explain this finding. The data suggest that leukocytes from BBDR contain cells with the ability to regulate reactivated autoreactive T cells in an autoimmune environment. This in vivo model of recurrent diabetes can therefore be used to define which type of cells are most effective in suppressing established autoimmune destruction of beta-cells.