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.
Highlights
Adenovirus (HAdV), cytomegalovirus (CMV), Epstein-BarrVirus (EBV), and polyoma-Virus (BKV) are responsible for serious morbidity and mortality in patients after hematopoietic stem cell transplantation (HSCT) [1,2,3,4]
PCR screening for human adenovirus (HAdV) following allogeneic HSCT allows early detection of impending invasive HAdV-infections, and timely preemptive antiviral treatment [8,36]
Few groups suggested that combined monitoring of viral load and virus-specific immunity by ELIspot, tetramer staining or the IFNc-cytokine secretion assay (CSA) has a clinical impact on the therapeutic intervention for pediatric allogeneic HSCT patients [36,37,38,39]
Summary
Adenovirus (HAdV), cytomegalovirus (CMV), Epstein-BarrVirus (EBV), and polyoma-Virus (BKV) are responsible for serious morbidity and mortality in patients after hematopoietic stem cell transplantation (HSCT) [1,2,3,4]. Certain sequences of the major capsid protein hexon are highly conserved among human HAdV which currently comprise more than 55 sybtypes divided into 7 different species (A–G)[23] This provides the basis for ‘‘cross-reactivity’’ of HAdV-specific T-cells facilitating broad recognition and protection against several species [24]. Only few HAdV-specific immunodominant CD8+ T-cell epitopes have been identified that are presented in the context of the common HLA-types A*01, A*24, B*07 and B*35 [14,27] greatly limiting the number of available HAdV-multimers. Using these four multimers, the probability to detect ADV-specific Tcells within the Caucasian population is about 73%. Our primary aim was to identify new promising ADV-specific epitopes for the HLA-types A*01 and A*24, and for the frequent HLA-type A*02, by analyzing the main structural proteins of the virus, including hexon and protein II, as well as the E1A protein expressed very early after infection
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