Exploiting cytokine secretion to rapidly produce multivirus-specific T cells for adoptive immunotherapy

J Immunother. 2008 Sep;31(7):665-74. doi: 10.1097/CJI.0b013e318181b4bd.


Viral infections remain a major cause of morbidity and mortality after hematopoietic stem cell transplantation (HSCT), and conventional small-molecule therapeutics often have modest benefit, high cost, and adverse effects. Adoptive transfer of donor-derived virus-specific T cells has proved feasible and safe after HSCT and to reconstitute immunity against cytomegalovirus, Epstein-Barr virus, and adenovirus. Current protocols to generate these cytotoxic T cell lines are lengthy, taking up to 12 weeks. As viral infections often occur <30 days after HSCT, speedy production of virus-specific cytotoxic T cells lacking alloreactivity is highly desirable. We now describe a modified rapid selection method for production and characterization of CD4 and CD8 T cells specific for cytomegalovirus, Epstein-Barr virus, and adenovirus in a single infusate. We use Ad5f35-pp65/latent membrane protein 2 vectors in a single procedure over a 48-hour time period and manufacture a product suited for clinical use. By simultaneously expanding a portion of the selected product, we can characterize phenotype and function of the infused product and link them with subsequent in vivo outcome.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviruses, Human / immunology
  • Cell Proliferation
  • Cells, Cultured
  • Cytomegalovirus / immunology
  • DNA Virus Infections / immunology
  • DNA Virus Infections / prevention & control*
  • Genetic Vectors*
  • Hematopoietic Stem Cell Transplantation
  • Herpesvirus 4, Human / immunology
  • Humans
  • Immunophenotyping
  • Immunotherapy, Adoptive*
  • Interferon-gamma / immunology*
  • Lymphocyte Activation / immunology
  • Phosphoproteins / immunology
  • T-Cell Antigen Receptor Specificity / immunology
  • T-Lymphocytes, Cytotoxic / immunology*
  • Viral Matrix Proteins / immunology


  • EBV-associated membrane antigen, Epstein-Barr virus
  • Phosphoproteins
  • Viral Matrix Proteins
  • cytomegalovirus matrix protein 65kDa
  • Interferon-gamma