Towards gene therapy for EBV-associated posttransplant lymphoma with genetically modified EBV-specific cytotoxic T cells

Blood. 2014 Oct 16;124(16):2514-22. doi: 10.1182/blood-2014-01-553362. Epub 2014 Sep 2.

Abstract

Epstein-Barr virus (EBV)-associated posttransplant lymphoma (PTLD) is a major cause of morbidity/mortality after hematopoietic stem cell (SCT) or solid organ (SOT) transplant. Adoptive immunotherapy with EBV-specific cytotoxic lymphocytes (CTLs), although effective in SCT, is less successful after SOT where lifelong immunosuppression therapy is necessary. We have genetically engineered EBV-CTLs to render them resistant to calcineurin (CN) inhibitor FK506 through retroviral transfer of a calcineurin A mutant (CNA12). Here we examined whether or not FK506-resistant EBV-CTLs control EBV-driven tumor progression in the presence of immunosuppression in a xenogeneic mouse model. NOD/SCID/IL2rγ(null) mice bearing human B-cell lymphoma were injected with autologous CTLs transduced with either CNA12 or eGFP in the presence/absence of FK506. Adoptive transfer of autologous CNA12-CTLs induced dramatic lymphoma regression despite the presence of FK506, whereas eGFP-CTLs did not. CNA12-CTLs persisted longer, homed to the tumor, and expanded more than eGFP-CTLs in mice treated with FK506. Mice receiving CNA12-CTLs and treated with FK506 survived significantly longer than control-treated animals. Our results demonstrate that CNA12-CTL induce regression of EBV-associated tumors in vivo despite ongoing immunosuppression. Clinical application of this novel approach may enhance the efficacy of adoptive transfer of EBV-CTL in SOT patients developing PTLD without the need for reduction in immunosuppressive therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcineurin / genetics
  • Calcineurin Inhibitors / pharmacology
  • Drug Resistance
  • Epstein-Barr Virus Infections / complications*
  • Genetic Engineering / methods
  • Genetic Therapy* / methods
  • Herpesvirus 4, Human / genetics
  • Herpesvirus 4, Human / isolation & purification
  • Humans
  • Immunosuppressive Agents / pharmacology
  • Immunotherapy, Adoptive* / methods
  • Lymphoma / genetics
  • Lymphoma / pathology
  • Lymphoma / therapy*
  • Lymphoma / virology*
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Mutation
  • T-Lymphocytes, Cytotoxic / drug effects
  • T-Lymphocytes, Cytotoxic / metabolism
  • T-Lymphocytes, Cytotoxic / transplantation*
  • T-Lymphocytes, Cytotoxic / virology*
  • Tacrolimus / pharmacology
  • Transduction, Genetic

Substances

  • Calcineurin Inhibitors
  • Immunosuppressive Agents
  • Calcineurin
  • Tacrolimus