Fig. 4.
Expression, cytotoxicity, and proliferation of scFv-CD28-ζ–transduced T cells from WT, IFN-γ−/−, pfp−/−, and pfp−/−IFN-γ−/−mice.

Expression, cytotoxicity, and proliferation of scFv-CD28-ζ–transduced T cells from WT, IFN-γ−/−, pfp−/−, and pfp−/−IFN-γ−/−mice.

(A) Equivalent expression of the scFv-anti-erbB2-CD28-ζ chimeric receptor in primary mouse T lymphocytes from WT, IFN-γ−/−, pfp−/−, and pfp−/−IFN-γ−/− BALB/c mice as determined by flow cytometry. Cells were stained with an anti–tag mAb (solid-line histogram) or an IgG1 isotype control mAb (dashed-line histogram). (B) The cytolytic activity of the scFv-anti-erbB2-CD28-ζ–transduced T cells from WT and gene-targeted mice was evaluated in a 6-hour51Cr release assay. Lysis of MC-38-erbB2+ tumor cells (closed symbols) by transduced IFN-γ−/− T cells (circles), but not pfp−/− (triangles) or pfp−/−INF-γ−/− T cells (diamonds), was equivalent to that mediated by gene-modified WT T cells (squares). All transduced effector T-cell populations were unable to lyse the erbB2MC-38 parental tumor cells (open symbols). Results from a representative experiment are expressed as percent specific51Cr release ± SE of triplicate samples. Spontaneous lysis was consistently less than 10%. (C) The proliferative capacity of scFv-anti-erbB2-CD28-ζ–transduced mouse T lymphocytes from WT (striped bars), pfp−/− (dotted bars), IFN-γ−/− (open bars), and pfp−/−IFN-γ−/− (closed bars) mice was determined by [3H]-thymidine incorporation at 72 hours. The ability of receptor-modified T cells from gene-targeted mice to proliferate in response to erbB2 antigen or antibody-mediated crosslinking of endogenous CD3 and CD28 receptors was comparable to that observed for transduced WT T cells. All transduced T-cell populations failed to proliferate in response to erbB2MC-38 parental tumor cells. Incorporation of radioactivity by tumor cells was not detected. Results from a representative experiment are expressed as means ± SE of triplicate samples.

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