title: Generation of a multipathogen-specific T-cell product for adoptive immunotherapy based on activation-dependent expression of CD154.
abstract: 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.
authors.affiliation: Department of Biomedicine and Division of Infectious Diseases and Hospital Epidemiology, University Hospital of Basel, Basel, Switzerland.
authors.forename: Nina
authors.initials: N
authors.lastname: Khanna
authors.forename: Claudia
authors.initials: C
authors.lastname: Stuehler
authors.forename: Barbara
authors.initials: B
authors.lastname: Conrad
authors.forename: Sarah
authors.initials: S
authors.lastname: Lurati
authors.forename: Sven
authors.initials: S
authors.lastname: Krappmann
authors.forename: Hermann
authors.initials: H
authors.lastname: Einsele
authors.forename: Carsten
authors.initials: C
authors.lastname: Berges
authors.forename: Max S
authors.initials: MS
authors.lastname: Topp
mesh_terms: D000256:Adenoviridae / Q000276:immunology; D000946:Antigens, Fungal / Q000276:immunology*; D000956:Antigens, Viral / Q000276:immunology*; D001232:Aspergillus fumigatus / Q000276:immunology; D023201:CD40 Ligand / Q000096:biosynthesis / Q000276:immunology*; D002176:Candida albicans / Q000276:immunology; D002460:Cell Line; D002469:Cell Separation; D003587:Cytomegalovirus / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D004854:Herpesvirus 4, Human / Q000276:immunology; D006801:Humans; D016130:Immunophenotyping; D016219:Immunotherapy, Adoptive / Q000379:methods*; D008213:Lymphocyte Activation / Q000276:immunology; D007959:Lymphocyte Culture Test, Mixed; D009181:Mycoses / Q000517:prevention & control; D013601:T-Lymphocytes / Q000276:immunology*; D014777:Virus Diseases / Q000517:prevention & control
chemical_list: D000946:Antigens, Fungal; D000956:Antigens, Viral; D023201:CD40 Ligand