Fig. 3.

CD8 T cell clone recognition of endogenously presented glutamic acid decarboxylase 65 (GAD65) by transduced target cells. CD8 T cell clones were co-cultured with GAD2 transduced human leucocyte antigen (HLA)-A2 expressing K562 cells (K562-A2-GAD65) for 18 h, and supernatants assessed for macrophage inflammatory protein (MIP)-1α and MIP-1β production. Both RK9P9-3 and RK9C10-1 respond similarly to stimulation with GAD9-mer peptide-pulsed cells. Similarly, neither respond to the empty vector control cell line. RK9C10-1 produced high levels of chemokines when cultured with K562-A2-GAD65 target cells, but RK9P9-3 did not respond. These data indicate that only RK9C10-1 recognizes GAD65 after endogenous processing, and that the naturally processed and presented epitope is GAD65_114–123. Cells were incubated at an effector : target ratio of 1:1. Bars show triplicate means and error bars standard errors of the mean of representative experiments.