Figure 4.

Clonotypic analysis of CD8⁺ T cells generated with mock-transfected and SOCS1-silenced DCs. CD8⁺ T cells from HLA-A*0201⁺ HIV-seronegative donors were primed and expanded with MDDCs that were either mock-transfected or treated with SOCS-1 siRNA. CD8⁺pentamer⁺ T cells were sorted by flow cytometry to >98% purity and all the expressed TRB gene products were analyzed using an unbiased template-switch anchored RT-PCR. CDR3 amino acid sequences, TRBV and TCRBJ usage, and the relative frequency of CD8⁺ T cell clonotypes specific for the (a) Melan-A/MART-1 and (b) HIV Gag TV9 epitopes are shown. Analyses were conducted on day 9 in (a) or on days 9 and day 23 in (b); top and bottom rows, respectively. Shaded boxes indicate common clonotypes. The IMGT nomenclature is used.⁴⁸ DC, dendritic cell; HIV, human immunodeficiency virus; IFN-γ, interferon-γ MDDC, myeloid-derived DC; RT-PCR, real-time PCR; SOCS-1, suppressor of cytokine signaling-1; TCR, T-cell receptor.