Wilson’s main area of focus will be on the HIV-1 vaccine design, specifically by targeting the immune response towards vulnerable regions in which escape mutations may significantly attenuate HIV. This may result in limited immune escape or in the event of escape, a less fit virus. HIV-1 Nef is a highly variable protein with many HLA-associated mutations potentially conferring escape from CD8+ T cell responses. Identification of immune escape mutations influencing pathogenic functions of Nef may indicate Nef regions that could be included in an HIV-1 vaccine. HIV Nef is an attractive target for vaccine design given its important role in enhancing viral replication and pathogenesis. The functional consequences of these mutations require further confirmation by site-directed mutagenesis experiments. The project aims at investigating the effect of these identified HLA-associated mutations on Nef function. The relevant Nef mutations will be introduced into three different patient-derived Nef clones that have high similarity to the consensus C Nef sequence. A CD4+ T cell line will then be transfected with the mutant Nef clones and the CD4 and HLA-I expression will be subsequently measured by flow cytometry to assess the CD4 and HLA-I down-regulation ability of the mutant Nef proteins.