Type 1 diabetes (T1D) results from the autoimmune destruction of pancreatic β cells. T cells (both CD8+ and CD4+) are critical to T1D progression and pathogenic T cell responses are well established at disease diagnosis. In mouse models, CD8+ T cells directly destroy β cells. In order for an immunotherapy to be effective, diabetogenic CD8+ T cell responses must be terminated. Genetically targeting antigen expression to antigen-presenting cells induces antigen-specific tolerance in CD8+ T cells. As insulin has been shown to be the primary antigen, we want to determine whether immunological tolerance can be induced in a spontaneous diabetes model, using insulin as the tolerogenic antigen. When insulin-specific CD8+ (G9C8) T cells were transferred to non-transgenic mice, G9C8 cells proliferated only in pancreatic lymph nodes (LN) where insulin is normally presented. In contrast, G9C8 cells proliferated in spleen, skin-draining LN and pancreatic LN of transgenic mice expressing proinsulin. Despite this, there was no difference in the number of G9C8 cells recovered from lymphoid tissues after transfer to proinsulin transgenic and non-transgenic mice. This was not due to site-specific trafficking as G9C8 cells did not accumulate in the pancreas of proinsulin transgenic or non-transgenic mice within 3 days of transfer. However, G9C8 cells recovered from proinsulin transgenic mice display a predominantly activated, ‘central memory’ phenotype. Finally, G9C8 T cell receptor expression was reduced in proinsulin transgenic but not non-transgenic recipients. These data suggest G9C8 cells are susceptible to antigen-specific tolerance induction despite inherent genetic defects that exist in NOD mice.