title: T cell re-targeting to EBV antigens following TCR gene transfer: CD28-containing receptors mediate enhanced antigen-specific IFNgamma production.
abstract: EBV is associated with a broad range of malignancies. Adoptive immunotherapy of these tumors with EBV-specific CTL proved useful. We generated a panel of primary human T cells specific to various EBV antigens (i.e. Epstein-Barr nuclear antigen 3A, 3B and BamHI-M leftward reading frame) via transfer of modified TCR genes that are either coupled to CD3zeta or Fc(epsilon)RIgamma. TCR-transduced T cells from 20-60% of donors (total number of 25) demonstrated specific lysis of EBV peptide-loaded target cells, whereas lymphoblastoid cell lines expressing native EBV antigens were not killed by any of the EBV-specific T cell populations. This non-responsiveness, confirmed at the level of nuclear factor of activated T cells activation, is not due to receptor configuration since identical receptor formats specific for melanoma antigens successfully re-targeted T cells to native melanoma cells. In an effort to generate a more potent receptor, we introduced a CD28 domain into one of the EBV-specific TCR. This TCR did not affect the cytotoxic response of re-targeted T cells, but dramatically enhanced antigen-specific IFNgamma production. We therefore conclude that these novel CD28-containing EBV-specific TCRs provide a basis for further development of TCR gene transfer to treat EBV-induced diseases.
authors.affiliation: Laboratory of Tumor Immunology, Department of Medical Oncology, Erasmus MC-Daniel den Hoed Cancer Center, Groene Hilledijk 301, 3075 EA Rotterdam, the Netherlands.
authors.forename: Niels
authors.initials: N
authors.lastname: Schaft
authors.forename: Birgit
authors.initials: B
authors.lastname: Lankiewicz
authors.forename: Joost
authors.initials: J
authors.lastname: Drexhage
authors.forename: Cor
authors.initials: C
authors.lastname: Berrevoets
authors.forename: Denis J
authors.initials: DJ
authors.lastname: Moss
authors.forename: Victor
authors.initials: V
authors.lastname: Levitsky
authors.forename: Marc
authors.initials: M
authors.lastname: Bonneville
authors.forename: Steven P
authors.initials: SP
authors.lastname: Lee
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: McMichael
authors.forename: Jan-Willem
authors.initials: JW
authors.lastname: Gratama
authors.forename: Reinier L H
authors.initials: RL
authors.lastname: Bolhuis
authors.forename: Ralph
authors.initials: R
authors.lastname: Willemsen
authors.forename: Reno
authors.initials: R
authors.lastname: Debets
mesh_terms: D018106:CD28 Antigens / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D019309:Epstein-Barr Virus Nuclear Antigens / Q000276:immunology*; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D019672:Genes, T-Cell Receptor; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis*; D019169:Jurkat Cells; D011948:Receptors, Antigen, T-Cell / Q000235:genetics / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*; D014161:Transduction, Genetic
chemical_list: D018106:CD28 Antigens; D019309:Epstein-Barr Virus Nuclear Antigens; D011948:Receptors, Antigen, T-Cell; D007371:Interferon-gamma