Representative analysis of TCRBV usage in CD8 T cells responding to HIV-infected targets. (a) Agarose gel analysis of anchored PCR amplification of TCRBV genes from immunomagnetically captured IFN-γ+ cells stimulated with autologous virus from representative subject 307. TCRBV and CD3 region sequencing of 50 cloned PCR products revealed a single 7S1 clonotype (see Table 2). Lanes depict molecular weight markers (M), negative control containing no cDNA template (1), and TCRBV gene amplification product from autologous virus–responding CD8 T cells (2). (b) Independent evaluation of TCRBV gene usage by PCR amplification of the same sample with a panel of 5′ Vβ primers and a common 3′ Cβ primer (lane numbers indicate individual Vβ families and decimals indicate subfamilies). (c) Flow cytometric analysis of cultured IFN-γ+ CD8 T cells from the same donor stained with TCRBV antibody corresponding to TCRBV gene 7S1.