The use of Chimeric antigen receptor (CAR) T cells is extremely effective in many haematological cancers, however to date, efficacy in solid tumours has proven less successful. Clinical trials report that high numbers of T cells infused into patients correlates with potential life threatening side effects, and therefore it is of benefit to infuse T cells with the potential for greater survival, proliferative capacity, and effector function in vivo. The clinical production of CAR-T cells requires an ex vivo T cell activation step using aCD3, aCD28 and IL-2, signals which program the T cell fate. Here, we have investigated the role for additional co-stimulatory signals to program CD8 T cell fates as measured by survival, proliferation, and effector function. Transgenic peptide-stimulated OT-I T cells were activated in the presence of various co- stimulatory agonists, with and without IL-2. Cell survival and division was measured over time using division tracking dye, before assaying for cell function. Our data demonstrates that alternative co-stimulation signalling induced unique profiles of survival, division potential, cytokine production and cell cytotoxicity in a dose dependent manner. Our data reveal that the initial priming of CD8+ T cells can drastically influence cell fate and persistence. With a better understanding of the biology behind how a T cell integrates co- stimulatory signals during priming we may be able to ‘tailor’ design current clinical practice to informatively predict the T cell response. We anticipate that this research may be applicable to the CAR-T cell immunotherapy field, with a potential to influence clinical practice.