Flow cytometry analysis of transduced T cells.

<p>A, analysis of the TCR-transduced EZP cell lines for CM9 peptide/MHC tetramer and SL8 peptide/MHC tetramer is presented with that of the untransduced CD8⁺ control cell line from recipient animal EZP. B, tetramer analysis of two SIV-specific CTL clones isolated from donor animal DAJ is presented above tetramer-sorted TCR transduced CD8⁺ cell lines. The DAJ SL8–42 clone is the TCR gene donor for the SL8–42 TCR EZP cell line.</p

Diagram of TCR expressing retroviral vector and the mature TCR chains.

<p>The MSGV1 murine retroviral vectors sequences are displayed as black boxes and lines while the TCR expression cassette is in white. The fine structure of the TCR chain fusion cassette is presented below the vector with the different MamuA*01-restricted TCRs molecularly-cloned from DAJ T-cell clones that were inserted into the vectors indicated above the cassette. The sequences separating the TCR genes, the furin recognition sequence, KAKR, the S-G-S-G spacer, and the P2A fowl pox self-cleaving peptide, are shaded gray. The furin cleavage site and the P2A self-cleavage site are indicated below the cassette with arrows. The mature α and β chains produced by this vector are displayed at the bottom of the figure.</p

In <i>vitro</i> virus suppression assay of TCR-transduced cell lines.

<p>Flow cytometry analyses of mixed cultures consisting of effector CD8⁺ T-cell lines and a target autologous CD4⁺ T-cell clone that was untreated or exposed to either wild-type SIV_mac239 or SIV_myr- are presented. Effectors are labeled above each column and targets are labeled at the left of each row. The effector CD8⁺ T cells in the co-cultures were stained with CellTrace Violet® and excluded from the analysis so that only the target cells were counted.</p