Expression of the chimeric scFv-anti-erbB2-ζ and scFv-anti-erbB2-CD28-ζ receptors in mouse T lymphocytes.

(A) Schematic representation of the scFv-anti-erbB2-ζ and scFv-anti-erbB2-CD28-ζ receptors. Each construct was composed of the V_H and V_L regions of the anti-erbB2 mAb joined by a flexible linker, a membrane-proximal hinge region, and the transmembrane (TM) and cytoplasmic regions of the human TCR-ζ chain or the mouse CD28 signaling chain fused to the intracellular domain of ζ. A c-myc tag epitope was incorporated into the C-terminus of the V_L region for expression analysis. (B) Expression of the scFv-anti-erbB2-ζ and anti-erbB2-CD28-ζ chimeric receptors in primary mouse T lymphocytes was analyzed by flow cytometry. Cells were stained with an anti–tag mAb or an IgG1 isotype control mAb. No expression was detected on T cells transduced with the retroviral vector, pLXSN, alone (bottom panel). Solid-line histogram depicts T cells stained with the anti–tag mAb; dashed-line histogram, T cells stained with IgG1 isotype control mAb.