Generation of Epstein-Barr virus-specific cytotoxic T lymphocytes resistant to the immunosuppressive drug tacrolimus (FK506).
Adoptive transfer of autologous Epstein-Barr virus-specific cytotoxic T lymphocytes (EBV-CTLs) to solid organ transplant (SOT) recipients has been shown safe and effective for the treatment of EBV-associated posttransplantation lymphoproliferative disorders (PTLDs). SOT recipients, however, require the continuous administration of immunosuppressive drugs to prevent graft rejection, and these agents may significantly limit the long-term persistence of transferred EBV-CTLs, precluding their use as prophylaxis. Tacrolimus (FK506) is one of the most widely used immunosuppressive agents in SOT recipients, and its immunosuppressive effects are largely dependent on its interaction with the 12-kDa FK506-binding protein (FKBP12). We have knocked down the expression of FKBP12 in EBV-CTLs using a specific small interfering RNA (siRNA) stably expressed from a retroviral vector and found that FKBP12-silenced EBV-CTLs are FK506 resistant. These cells continue to expand in the presence of the drug without measurable impairment of their antigen specificity or cytotoxic activity. We confirmed their FK506 resistance and anti-PTLD activity in vivo using a xenogenic mouse model, suggesting that the proposed strategy may be of value to enhance EBV-specific immune surveillance in patients at high risk of PTLD after transplantation.
De Angelis
Biagio
B
Center for Cell and Gene Therapy, Baylor College of Medicine, The Methodist Hospital and Texas Children's Hospital, Houston, TX 77030, USA.
Dotti
Gianpietro
G
Quintarelli
Concetta
C
Huye
Leslie E
LE
Zhang
Lan
L
Zhang
Ming
M
Pane
Fabrizio
F
Heslop
Helen E
HE
Brenner
Malcolm K
MK
Rooney
Cliona M
CM
Savoldo
Barbara
B
D019264:Adoptive Transfer; D000818:Animals; D000956:Antigens, Viral / Q000276:immunology; D002478:Cells, Cultured / Q000187:drug effects / Q000276:immunology / Q000637:transplantation; D003602:Cytotoxicity, Immunologic; D004351:Drug Resistance / Q000235:genetics; D055785:Gene Knockdown Techniques; D005822:Genetic Vectors / Q000494:pharmacology*; D004854:Herpesvirus 4, Human / Q000276:immunology*; D006801:Humans; D007166:Immunosuppressive Agents / Q000627:therapeutic use*; D008228:Lymphoma, Non-Hodgkin / Q000628:therapy / Q000821:virology; D051379:Mice; D016513:Mice, SCID; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000037:antagonists & inhibitors; D034741:RNA, Small Interfering / Q000502:physiology*; D011993:Recombinant Fusion Proteins / Q000037:antagonists & inhibitors / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D019893:Ribosomal Protein S6 Kinases / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects* / Q000276:immunology / Q000637:transplantation; D016559:Tacrolimus / Q000494:pharmacology*; D022061:Tacrolimus Binding Protein 1A / Q000037:antagonists & inhibitors* / Q000187:drug effects / Q000235:genetics; D023041:Xenograft Model Antitumor Assays
D000956:Antigens, Viral; D007166:Immunosuppressive Agents; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D019893:Ribosomal Protein S6 Kinases; D022061:Tacrolimus Binding Protein 1A; D016559:Tacrolimus