The capacity of Coxsackie B viruses (CBVs) to produce diabetes in mice was studied before and after passage in various cell types. CBVs that had been passaged in monkey kidney cells or in mouse embryo fibroblasts failed to produce abnormal glucose tolerance tests, whereas virus passaged five or more times in the pancreata of mice or in beta-cell cultures produced transient abnormal glucose tolerance tests. Immunofluorescence and histologic studies revealed that passage of CBVs in cultured beta-cells changed the tropism of these viruses from the acinar pancreas to the islets of Langerhans. Although all six CBV serotypes that had been passaged in beta-cell cultures behaved very similarly, substantial variation was observed with the different virus passages and in some experiments, beta-cell damage and the glucose abnormalities were minimal. From these and other experiments, we conclude that the six members of the CBV group have the potential for infecting and damaging pancreatic beta-cells in mice.