Inheritance of Type 1 diabetes (T1D) associated-risk alleles at certain genetic loci is unable to completely account for either the onset or acceleration of Type 1 diabetes (T1D). Increasing evidence suggests that environmental factors are key triggers that contribute to disease onset. Notably, in a recent study, infection with the Coxsackievirus B1 (CBV1) serotype has been linked to T1D progression in a large prospective birth cohort study¹ . CD8 T lymphocytes represent a major component of anti-viral adaptive immune responses, but are poorly understood in relation to enterovirus infection and T1D. To address this, we used in silico peptide prediction algorithms to generate a candidate list of 107 peptides with high predicted scores for presentation by HLA-A*0201. We then used sensitive CD8 IFN-gamma ELISPOT studies on CBV1-seropositive donors to screen these candidates in peptide pools and identify several novel candidate CD8 T cell peptide targets derived from CBV1. Following co-culture of peptide-pulsed dendritic cells we generated CD8 T cell lines and subsequently clones directed against these putative targets. Thus our study confirms the presence of peripheral CBV1-specific effector CD8 T cells directed against novel, CVB1-restricted epitopes in CVB1-experienced subjects. CBV1-specific CD8 T cell clones will elucidate the natural processing of these by target cells and enable peptide-HLA multimer studies to monitor peripheral numbers of circulating CBV1-specific CD8 T cells for the first time.