title: Redirected perforin-dependent lysis of colon carcinoma by ex vivo genetically engineered CTL.
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.
authors.affiliation: Cellular Cytotoxicity Laboratory, The Austin Research Institute, Heidelberg, Victoria, Australia. p.darcy@ari.unimelb.edu.au
authors.forename: P K
authors.initials: PK
authors.lastname: Darcy
authors.forename: N M
authors.initials: NM
authors.lastname: Haynes
authors.forename: M B
authors.initials: MB
authors.lastname: Snook
authors.forename: J A
authors.initials: JA
authors.lastname: Trapani
authors.forename: L
authors.initials: L
authors.lastname: Cerruti
authors.forename: S M
authors.initials: SM
authors.lastname: Jane
authors.forename: M J
authors.initials: MJ
authors.lastname: Smyth
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000276:immunology / Q000517:prevention & control; D019264:Adoptive Transfer; D000818:Animals; D001665:Binding Sites / Q000235:genetics / Q000276:immunology; D002272:Carcinoembryonic Antigen / Q000378:metabolism; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology; D002455:Cell Division / Q000276:immunology; D003110:Colonic Neoplasms / Q000235:genetics* / Q000276:immunology* / Q000473:pathology / Q000517:prevention & control; D016207:Cytokines / Q000378:metabolism; D003602:Cytotoxicity, Immunologic / Q000235:genetics*; D018984:Epitopes, T-Lymphocyte / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007128:Immunoglobulin Fragments / Q000096:biosynthesis / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics; D007371:Interferon-gamma / Q000502:physiology; D018655:Lymphocyte Count; D008562:Membrane Glycoproteins / Q000235:genetics / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D016513:Mice, SCID; D009368:Neoplasm Transplantation; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000138:chemical synthesis / Q000235:genetics; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000378:metabolism / Q000637:transplantation; D014161:Transduction, Genetic; D014407:Tumor Cells, Cultured
chemical_list: D002272:Carcinoembryonic Antigen; D002352:Carrier Proteins; D016207:Cytokines; D018984:Epitopes, T-Lymphocyte; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; C065686:carcinoembryonic antigen binding protein, human; C055966:immunoglobulin Fv; D054353:Perforin; D007371:Interferon-gamma