title: Rapid generation of EBV-specific cytotoxic T lymphocytes resistant to calcineurin inhibitors for adoptive immunotherapy.
abstract: Epstein-Barr virus (EBV)-associated posttransplant lymphoproliferative disorder (PTLD) remains a major cause of morbidity and mortality after hematopoietic stem cell (HSCT) or solid organ transplant (SOT). Strategies to reconstitute immunity by adoptive transfer of EBV-specific cytotoxic T lymphocyte (CTL) therapy while highly effective in the HSCT setting where immunosuppression can be withdrawn have been less successful in the SOT setting where continued immunosuppression therapy is necessary. Additionally, the complexity and time taken to generate EBV-CTLs for adoptive transfer limit the clinical applicability. We have developed a system for the rapid generation of EBV-CTLs resistant to immunosuppression based on selection of interferon-gamma (IFN-γ) secreting EBV-CTLs and retroviral transduction with a calcineurin B mutant. With this methodology, EBV-CTLs resistant to the calcineurin inhibitor Tacrolimus (TAC) can be produced in 14 days. These CTLs show high specificity for EBV with negligible alloreactivity in both proliferation and cytotoxicity assays and are able to proliferate and secrete IFN-γ in response to antigen stimulation in the presence of therapeutic doses of TAC. This strategy will substantially facilitate clinical application of this approach for the treatment of PTLD in SOT recipients.
authors.affiliation: Molecular Immunology Unit, Institute of Child Health, UCL, London, UK; Department of Haematology, UCL Cancer Institute, London, UK.
authors.forename: I
authors.initials: I
authors.lastname: Ricciardelli
authors.forename: J
authors.initials: J
authors.lastname: Brewin
authors.forename: G
authors.initials: G
authors.lastname: Lugthart
authors.forename: S J
authors.initials: SJ
authors.lastname: Albon
authors.forename: M
authors.initials: M
authors.lastname: Pule
authors.forename: P J
authors.initials: PJ
authors.lastname: Amrolia
mesh_terms: D000941:Antigens / Q000276:immunology; D019703:Calcineurin / Q000235:genetics; D065095:Calcineurin Inhibitors; D049109:Cell Proliferation; D020031:Epstein-Barr Virus Infections / Q000276:immunology*; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects; D004854:Herpesvirus 4, Human; D006801:Humans; D007156:Immunologic Memory; D007165:Immunosuppression Therapy; D016219:Immunotherapy, Adoptive; D007371:Interferon-gamma / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000166:cytology; D008223:Lymphoma / Q000276:immunology*; D009154:Mutation; D016377:Organ Transplantation / Q000009:adverse effects; D010641:Phenotype; D011183:Postoperative Complications; D012190:Retroviridae / Q000378:metabolism; D013601:T-Lymphocytes / Q000821:virology; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology*; D016559:Tacrolimus / Q000494:pharmacology
chemical_list: D000941:Antigens; D065095:Calcineurin Inhibitors; D007371:Interferon-gamma; D019703:Calcineurin; D016559:Tacrolimus
keywords: Calcineurin inhibitors resistance; Epstein-Barr virus; IFN-γ selection; cytotoxic T lymphocytes; posttransplant lymphoma; solid organ transplant