title: Clinical-scale generation of multi-specific anti-fungal T cells targeting Candida, Aspergillus and mucormycetes.
abstract: OBJECTIVE
  Invasive fungal infections, in particular, infections caused by Candida, Aspergillus and mucormycetes, are a major cause of morbidity and mortality in patients undergoing allogeneic hematopoietic stem cell transplantation. Adoptive transfer of donor-derived anti-fungal T cells shows promise to restore immunity and to offer a cure. Because T cells recognize only specific epitopes, the low rate of patients in which the causal fungal pathogen can be identified and the considerable number of patients with co-infection with several genera or species of fungi significantly limit the application of adoptive immunotherapy.
  
  
  METHODS
  Using the interferon-γ secretion assay, we isolated multi-specific human anti-fungal T cells after simultaneous stimulation with cellular extracts of Aspergillus fumigatus, Candida albicans and Rhizopus oryzae. Cells were phenotypically and functionally characterized by flow cytometry.
  
  
  RESULTS
  Of a total of 1.1 × 10(9) peripheral blood mononuclear cells, a median number of 5.2 × 10(7) CD3+ CD4+ T cells was generated within 12 days. This cell population consisted of activated memory TH1 cells and reproducibly responded to a multitude of Aspergillus spp., Candida spp. and mucormycetes with interferon-γ production. On re-stimulation, the generated T cells proliferated and enhanced anti-fungal activity of phagocytes and showed reduced alloreactivity compared with the original cell fraction.
  
  
  CONCLUSIONS
  Our rapid and simple method of simultaneously generating functionally active multi-specific T cells that recognize a wide variety of medically relevant fungi may form the basis for future clinical trials investigating adoptive immunotherapy in allogeneic hematopoietic stem cell transplantation recipients with invasive fungal infection.
authors.affiliation: Pediatric Hematology and Oncology, Johann Wolfgang Goethe University, Frankfurt, Germany.
authors.forename: Lars
authors.initials: L
authors.lastname: Tramsen
authors.forename: Stanislaw
authors.initials: S
authors.lastname: Schmidt
authors.forename: Halvard
authors.initials: H
authors.lastname: Boenig
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Latgé
authors.forename: Cornelia
authors.initials: C
authors.lastname: Lass-Flörl
authors.forename: Frauke
authors.initials: F
authors.lastname: Roeger
authors.forename: Erhard
authors.initials: E
authors.lastname: Seifried
authors.forename: Thomas
authors.initials: T
authors.lastname: Klingebiel
authors.forename: Thomas
authors.initials: T
authors.lastname: Lehrnbecher
mesh_terms: D000328:Adult; D001230:Aspergillus / Q000276:immunology / Q000302:isolation & purification* / Q000472:pathogenicity; D002175:Candida / Q000276:immunology / Q000302:isolation & purification* / Q000472:pathogenicity; D005260:Female; D005434:Flow Cytometry; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects / Q000401:mortality; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology* / Q000378:metabolism; D059425:Interferon-gamma Release Tests; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology; D008297:Male; D008875:Middle Aged; D012233:Rhizopus / Q000276:immunology / Q000302:isolation & purification* / Q000472:pathogenicity; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology*
chemical_list: D007371:Interferon-gamma