Short-term in-vitro expansion improves monitoring and allows affordable generation of virus-specific T-cells against several viruses for a broad clinical application

PLoS One. 2013 Apr 22;8(4):e59592. doi: 10.1371/journal.pone.0059592. Print 2013.

Abstract

Adenoviral infections are a major cause of morbidity and mortality after allogeneic hematopoietic stem cell transplantation (HSCT) in pediatric patients. Adoptive transfer of donor-derived human adenovirus (HAdV)-specific T-cells represents a promising treatment option. However, the difficulty in identifying and selecting rare HAdV-specific T-cells, and the short time span between patients at high risk for invasive infection and viremia are major limitations. We therefore developed an IL-15-driven 6 to 12 day short-term protocol for in vitro detection of HAdV-specific T cells, as revealed by known MHC class I multimers and a newly identified adenoviral CD8 T-cell epitope derived from the E1A protein for the frequent HLA-type A*02∶01 and IFN-γ. Using this novel and improved diagnostic approach we observed a correlation between adenoviral load and reconstitution of CD8(+) and CD4(+) HAdV-specific T-cells including central memory cells in HSCT-patients. Adaption of the 12-day protocol to good manufacturing practice conditions resulted in a 2.6-log (mean) expansion of HAdV-specific T-cells displaying high cytolytic activity (4-fold) compared to controls and low or absent alloreactivity. Similar protocols successfully identified and rapidly expanded CMV-, EBV-, and BKV-specific T-cells. Our approach provides a powerful clinical-grade convertible tool for rapid and cost-effective detection and enrichment of multiple virus-specific T-cells that may facilitate broad clinical application.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenoviridae / immunology*
  • Adenoviridae Infections / immunology
  • Adenoviridae Infections / virology
  • Antigens, Viral / immunology
  • CD4-Positive T-Lymphocytes / immunology
  • CD4-Positive T-Lymphocytes / virology*
  • CD8-Positive T-Lymphocytes / immunology
  • CD8-Positive T-Lymphocytes / virology*
  • Cell Culture Techniques / economics
  • Cell Proliferation
  • Cells, Cultured
  • Child
  • Child, Preschool
  • Cytotoxicity, Immunologic
  • Female
  • Hematopoietic Stem Cell Transplantation / adverse effects
  • Humans
  • Male
  • Phenotype
  • Transplantation, Homologous
  • Virus Cultivation / economics
  • Young Adult

Substances

  • Antigens, Viral

Grant support

This work was supported in part by grants from Eurostars Grant E! 5744 (FFG 829495), Clinical Trial Investigator Driven (CTID)-RG-Matthes and the German Children's Cancer Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. No additional external funding received for this study.