Abstract
The redirection of autologous lymphocytes to predefined tumor target Ags has considerable potential for the immunotherapeutic treatment of cancer; however, robust experimental systems for comparing various approaches have not been developed. Herein, we have generated a single chain variable domain anti-carcinoembryonic Ag (CEA) Fcepsilon receptor I gamma-chain fusion (scFv anti-CEA) receptor and demonstrated high-level expression of this chimeric receptor in naive mouse T lymphocytes by retroviral gene transduction. These gene-modified CTL were able to lyse CEA+ targets and secrete high levels of IFN-gamma following Ag stimulation. Depletion studies demonstrated that specific tumor cell cytotoxicity was mediated by gene-modified CD8+ T cells. Importantly, in increasingly stringent tests of efficacy in vivo, transduced CTL were sequentially shown to reject CEA+ colon carcinoma cells in a Winn assay and then reject established s.c. colon carcinoma in scid or syngeneic mice. Furthermore, using gene-targeted and scFv anti-CEA receptor-transduced donor CTL, perforin and IFN-gamma were demonstrated to be absolutely critical for the eradication of colon carcinoma in mice. In summary, we have developed a highly efficient gene transfer system for evaluating chimeric receptor expression in cytotoxic lymphocytes. This series of experiments has revealed the utility of scFv anti-CEA chimeras in providing mouse T cells the capacity to reject colon carcinoma in an Ag- and perforin-specific manner.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenocarcinoma / genetics
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Adenocarcinoma / immunology
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Adenocarcinoma / prevention & control
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Adoptive Transfer
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Animals
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Binding Sites / genetics
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Binding Sites / immunology
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Carcinoembryonic Antigen / metabolism
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Carrier Proteins / genetics
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Carrier Proteins / immunology
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Cell Division / immunology
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Colonic Neoplasms / genetics*
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Colonic Neoplasms / immunology*
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Colonic Neoplasms / pathology
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Colonic Neoplasms / prevention & control
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Cytokines / metabolism
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Cytotoxicity, Immunologic / genetics*
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Epitopes, T-Lymphocyte / immunology
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Epitopes, T-Lymphocyte / metabolism
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Humans
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Immunoglobulin Fragments / biosynthesis
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Immunoglobulin Fragments / genetics
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Immunoglobulin Variable Region / genetics
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Interferon-gamma / physiology
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Lymphocyte Count
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Membrane Glycoproteins / genetics
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Membrane Glycoproteins / physiology*
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Mice
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Mice, Inbred BALB C
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Mice, Inbred C57BL
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Mice, Knockout
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Mice, SCID
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Neoplasm Transplantation
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Perforin
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Pore Forming Cytotoxic Proteins
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Receptors, Cell Surface*
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Recombinant Fusion Proteins / biosynthesis
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Recombinant Fusion Proteins / chemical synthesis
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Recombinant Fusion Proteins / genetics
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T-Lymphocytes, Cytotoxic / immunology*
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T-Lymphocytes, Cytotoxic / metabolism
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T-Lymphocytes, Cytotoxic / transplantation
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Transduction, Genetic
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Tumor Cells, Cultured
Substances
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Carcinoembryonic Antigen
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Carrier Proteins
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Cytokines
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Epitopes, T-Lymphocyte
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Immunoglobulin Fragments
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Immunoglobulin Variable Region
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Membrane Glycoproteins
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Pore Forming Cytotoxic Proteins
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Receptors, Cell Surface
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Recombinant Fusion Proteins
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carcinoembryonic antigen binding protein, human
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immunoglobulin Fv
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Perforin
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Interferon-gamma