Figure 2: Strategy for the detection of immune responses against an artificial allogeneic HLA variant. Experimental outline for detecting the T cell response against an individual HLA class I molecule. Monocytes isolated from peripheral blood of a selected donor (here expressing HLA-B*44:02) are used for the generation of DCs. Mature DCs are transduced with lentiviral constructs encoding for an additional full length HLA-B*44 allele differing by a single AA at position 156. These DCs are co-cultured with freshly isolated autologous CD3+ T cells. The HLA mismatch effect is determined by monitoring the responses of the stimulated T cells using recombinant DCs expressing addition HLA-B*44 molecule as target cells.