Antiviral CD8+ T cells play an important role in initial control of HIV replication and presumably help delay the onset of AIDS in HIV infected individuals. In order to understand the role of antiviral CD8+ T cells in immune protection, it will be necessary to comprehensively characterize the specificities CD8+ T cells in HIV-infected patients. A major obstacle to this goal is the high degree of genetic variability among HIV clinical isolates from different individuals. Most current techniques to screen T cell specificities rely on reference strain antigens or synthetic peptides representing a consensus sequence. These methods may underestimate T cell response due to genetic divergence between the assay antigen and autologous virus. This proposal describes an approach, based on transfection of antigen presenting cells with synthetic mRNA, for efficient screening of patient CDS+ T cell responses to autologous HIV antigens. This method will be used to compare the T cell responses to autologous and consensus antigens in chronic HIV infection.
