An Epstein-Barr virus deletion mutant associated with fatal lymphoproliferative disease unresponsive to therapy with virus-specific CTLs

Blood. 2001 Feb 15;97(4):835-43. doi: 10.1182/blood.v97.4.835.


There is a growing interest in using antigen-specific T cells for the treatment of human malignancy. For example, adoptive transfer of Epstein-Barr virus (EBV)-specific cytotoxic T lymphocytes (CTLs) has been effective prophylaxis and treatment of EBV-associated lymphoproliferative disease in immunocompromised patients. For all immunotherapies, however, there has been a hypothetical concern that mutations in tumor-specific antigens may lead to tumor escape. We now demonstrate that such events may indeed occur, with lethal outcome. A patient who developed lymphoma after marrow transplantation received donor-derived, EBV-specific CTLs but died with progressive disease. The tumor cells proved substantially less sensitive to cytolysis than the EBV-transformed B-cell line used for CTL generation. The major cytolytic activity of the donor CTL was directed against 2 HLA-A11-restricted epitopes in the viral EBNA-3B antigen. Sequence analysis of this gene in the tumor virus revealed a 245-base pair deletion, which removed these 2 CTL epitopes. Hence, the viral antigen in the tumor had mutated in a way that allowed escape from CTLs. Analysis of EBV polymorphisms demonstrated that before CTL infusion, more than one virus was present, including a virus with wild-type EBNA-3B. After CTL infusion, only the virus with the EBNA-3B deletion could be detected, suggesting that the infused CTLs had selected a resistant strain in vivo. Such an occurrence, even when polyclonal CTL lines are used against genetically stable virus antigens, suggests that escape mutants may be a serious problem when CTL therapy is directed against more unstable tumor cell-derived targets.

Publication types

  • Case Reports
  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adolescent
  • Amino Acid Sequence
  • Antineoplastic Agents, Alkylating / therapeutic use
  • B-Lymphocytes / immunology
  • Bone Marrow Transplantation / adverse effects*
  • Cell Line, Transformed / immunology
  • Combined Modality Therapy
  • Cyclophosphamide / therapeutic use
  • Cytotoxicity, Immunologic
  • Disease Progression
  • Epstein-Barr Virus Infections / genetics
  • Epstein-Barr Virus Infections / immunology*
  • Epstein-Barr Virus Infections / transmission
  • Epstein-Barr Virus Infections / virology
  • Epstein-Barr Virus Nuclear Antigens / genetics
  • Epstein-Barr Virus Nuclear Antigens / immunology*
  • Fatal Outcome
  • Female
  • Genes, Viral*
  • HLA-A Antigens / immunology
  • HLA-A11 Antigen
  • Herpesvirus 4, Human / genetics*
  • Herpesvirus 4, Human / immunology
  • Humans
  • Immunodominant Epitopes / genetics
  • Immunodominant Epitopes / immunology*
  • Immunotherapy, Adoptive*
  • Lymphoma, B-Cell / genetics
  • Lymphoma, B-Cell / immunology*
  • Lymphoma, B-Cell / virology
  • Molecular Sequence Data
  • Polymerase Chain Reaction
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / therapy
  • Sequence Alignment
  • Sequence Deletion
  • Sequence Homology, Amino Acid
  • T-Lymphocytes, Cytotoxic / immunology*
  • T-Lymphocytes, Cytotoxic / transplantation
  • Tissue Donors
  • Transplantation, Homologous / adverse effects*
  • Viral Structural Proteins / genetics*


  • Antineoplastic Agents, Alkylating
  • EBNA-3B antigen
  • Epstein-Barr Virus Nuclear Antigens
  • HLA-A Antigens
  • HLA-A11 Antigen
  • Immunodominant Epitopes
  • Viral Structural Proteins
  • Cyclophosphamide