title: Generation of EBV-specific cytotoxic T cells that are resistant to calcineurin inhibitors for the treatment of posttransplantation lymphoproliferative disease.
abstract: Epstein-Barr virus (EBV)-driven posttransplantation lymphoproliferative disease (PTLD) is a serious complication of immunosuppression after either stem cell transplantation (SCT) or solid organ transplantation (SOT). Adoptive transfer of EBV-specific cytotoxic T lymphocytes (EBV-CTLs) is an effective prophylaxis and treatment for PTLD after SCT, but not for PTLD after SOT when pharmacologic immunosuppression cannot be discontinued. We report the generation of calcineurin (CN) mutants that render EBV-CTL resistant to the immunosuppressants tacrolimus (FK506) and cyclosporin A (CsA): mutant CNa12 confers resistance to CsA but not FK506, and mutant CNa22 confers resistance to FK506 but not CsA, whereas mutant CNb30 renders CTLs resistant to both calcineurin inhibitors. Untransduced EBV-CTLs do not proliferate in the presence of FK506/CsA. However, EBV-CTLs transduced with a retroviral vector coding for these mutants retain the ability to both proliferate and secrete normal levels of interferon-gamma in the presence therapeutic levels of FK506 (CNa12), CsA (CNa22), or both (CNb30). The cytotoxicity and phenotype of EBV-CTL lines were unaffected by expression of these mutant CNs. This approach should allow effective immunotherapy with EBV-CTLs in the SOT setting without risking the graft by reduction in immunosuppression, and represents a generic approach to improving immunotherapy in the face of immunosuppression.
authors.affiliation: Molecular Immunology Unit, Institute of Child Health, University College London, London, United Kingdom.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Brewin
authors.forename: Christoph
authors.initials: C
authors.lastname: Mancao
authors.forename: Karin
authors.initials: K
authors.lastname: Straathof
authors.forename: Helen
authors.initials: H
authors.lastname: Karlsson
authors.forename: Sujith
authors.initials: S
authors.lastname: Samarasinghe
authors.forename: Persis J
authors.initials: PJ
authors.lastname: Amrolia
authors.forename: Martin
authors.initials: M
authors.lastname: Pule
mesh_terms: D019264:Adoptive Transfer; D019703:Calcineurin / Q000737:chemistry / Q000235:genetics*; D065095:Calcineurin Inhibitors; D002478:Cells, Cultured / Q000187:drug effects / Q000276:immunology; D021984:Cyclophilin A / Q000378:metabolism; D016572:Cyclosporine / Q000494:pharmacology*; D003602:Cytotoxicity, Immunologic; D004351:Drug Resistance / Q000235:genetics; D020031:Epstein-Barr Virus Infections / Q000628:therapy*; D005822:Genetic Vectors / Q000494:pharmacology*; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D007371:Interferon-gamma / Q000378:metabolism; D019169:Jurkat Cells / Q000187:drug effects / Q000276:immunology; D008232:Lymphoproliferative Disorders / Q000628:therapy*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D016377:Organ Transplantation; D011183:Postoperative Complications / Q000628:therapy*; D011487:Protein Conformation; D025941:Protein Interaction Mapping; D011993:Recombinant Fusion Proteins / Q000037:antagonists & inhibitors / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism / Q000637:transplantation; D016559:Tacrolimus / Q000494:pharmacology*; D022061:Tacrolimus Binding Protein 1A / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D065095:Calcineurin Inhibitors; D007166:Immunosuppressive Agents; D011993:Recombinant Fusion Proteins; D007371:Interferon-gamma; D016572:Cyclosporine; D019703:Calcineurin; D021984:Cyclophilin A; D022061:Tacrolimus Binding Protein 1A; D016559:Tacrolimus