Prophylactic insulin can prevent diabetes in the BB rat. We evaluated its use to prevent adoptive transfer of diabetes by activated splenocytes. ConA-activated spleen cells from acutely diabetic BB rats were divided into two equal aliquots and injected intravenously in paired diabetes-prone BB rat littermates. At the time of cell injection, subcutaneous insulin injections (15 U.kg-1.day-1) were started in one of each pair (n = 21) of littermates, and the control littermates (n = 21) were injected with saline. The incidence of diabetes, observed 35 days after cell injection, was 95% in control rats compared with 29% in insulin-treated rats (P less than 0.05). To confirm the absence of diabetes, insulin was stopped in all nondiabetic insulin-treated rats at 63 days of age. An OGTT was performed at 65 days of age: 4 rats were glucose intolerant. All rats received comparable numbers of ConA-activated splenocytes. At the time the rats were killed, 3 insulin-treated rats had a completely normal morphology and a normal glucose tolerance. All control rats had insulitis whether they were diabetic or not. No significant difference in any mononuclear subset was observed in relation to insulin treatment. We conclude that prophylactic insulin can prevent adoptive transfer of diabetes in the BB rat without inducing changes in the mononuclear cell subsets.