title: Short-term in-vitro expansion improves monitoring and allows affordable generation of virus-specific T-cells against several viruses for a broad clinical application.
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.
authors: Department of Clinical Cell Biology and FACS Core Unit, Children's Cancer Research Institute (CCRI), Vienna, Austria. rene.geyeregger@ccri.at
authors: René
authors: R
authors: Geyeregger
authors: Christine
authors: C
authors: Freimüller
authors: Stefan
authors: S
authors: Stevanovic
authors: Julia
authors: J
authors: Stemberger
authors: Gabor
authors: G
authors: Mester
authors: Jasmin
authors: J
authors: Dmytrus
authors: Thomas
authors: T
authors: Lion
authors: Hans-Georg
authors: HG
authors: Rammensee
authors: Gottfried
authors: G
authors: Fischer
authors: Britta
authors: B
authors: Eiz-Vesper
authors: Anita
authors: A
authors: Lawitschka
authors: Susanne
authors: S
authors: Matthes
authors: Gerhard
authors: G
authors: Fritsch
mesh_terms: D000256:Adenoviridae / Q000276:immunology*; D000257:Adenoviridae Infections / Q000276:immunology / Q000821:virology; D000956:Antigens, Viral / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000821:virology*; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology / Q000821:virology*; D018929:Cell Culture Techniques / Q000191:economics; D049109:Cell Proliferation; D002478:Cells, Cultured; D002648:Child; D002675:Child, Preschool; D003602:Cytotoxicity, Immunologic; D005260:Female; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects; D006801:Humans; D008297:Male; D010641:Phenotype; D014184:Transplantation, Homologous; D014776:Virus Cultivation / Q000191:economics; D055815:Young Adult
chemical_list: D000956:Antigens, Viral
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