Generation of a multipathogen-specific T-cell product for adoptive immunotherapy based on activation-dependent expression of CD154

Blood. 2011 Jul 28;118(4):1121-31. doi: 10.1182/blood-2010-12-322610. Epub 2011 Jun 3.


Viral and fungal infections remain a leading cause of mortality in patients after hematopoietic stem cell transplantation (HSCT). Adoptive transfer of multipathogen-specific T cells is promising in restoring immunity and thereby preventing and treating infections, but approaches are currently limited because of time-consuming and laborious procedures. Therefore, we investigated a new strategy to simultaneously select T cells specific for viral and fungal pathogens based on activation-dependent expression of CD154. Single- and multipathogen-specific T-cell lines with high specificity for adenovirus (AdV), Epstein-Barr virus (EBV), cytomegalovirus (CMV), Candida albicans, and/or Aspergillus fumigatus could be readily generated within 14 days irrespective of the precursor frequency. The T-cell lines responded reproducibly to endogenously processed antigen and specifically proliferated upon antigenic stimulation. Although isolation based on CD154 favors enrichment of CD4(+) T cells, AdV-, EBV- and CMV-specific CD8(+) T cells could be expanded and demonstrated lysis of target cells. Conversely, T cell-mediated alloreactivity was almost abrogated compared with the starting fraction. This selection and/or expansion strategy may form the basis for future adoptive immunotherapy trials in patients at risk for multiple infections and may be translated to other antigens.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / immunology
  • Antigens, Fungal / immunology*
  • Antigens, Viral / immunology*
  • Aspergillus fumigatus / immunology
  • CD40 Ligand / biosynthesis
  • CD40 Ligand / immunology*
  • Candida albicans / immunology
  • Cell Line
  • Cell Separation
  • Cytomegalovirus / immunology
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Herpesvirus 4, Human / immunology
  • Humans
  • Immunophenotyping
  • Immunotherapy, Adoptive / methods*
  • Lymphocyte Activation / immunology
  • Lymphocyte Culture Test, Mixed
  • Mycoses / prevention & control
  • T-Lymphocytes / immunology*
  • Virus Diseases / prevention & control


  • Antigens, Fungal
  • Antigens, Viral
  • CD40 Ligand