Figure 4

Clonotype analysis of FLU- and GAG-stimulated lymphocytes. Some identical FLU-specific clonotypes between FLU- and GAG-stimulated T cells were observed in 5 BV families analyzed by denaturing gradient gel electrophoresis (DGGE). PBMC from a HIV-seronegative Donor 1 were stimulated in vitro with FLU-M1:58–66 or GAG:77–85 peptides, and the resulting FLU-M1:58–66-specific cells purified by binding cells with the cognate TCR to HLA-A2/FLU-M1:58–66 peptide monomers conjugated to magnetic microbeads. RT-PCR was performed on RNA derived from each cell type (FLU-M1:58–66- or GAG:77–85-stimulated) using primers specific for all 24 TCR-β (BV) families. Each band resolved on a DGGE gel represents different clonotypes separated according to their nucleotide composition. The results of five different primers are shown here (BV13a is one of the two variable primers needed to amplify all possible members of the BV13 family).