title: Robust polyfunctional T-helper 1 responses to multiple fungal antigens from a cell population generated using an environmental strain of Aspergillus fumigatus.
abstract: OBJECTIVE
  Aspergillus fumigatus infections are the leading cause of invasive fungal infection-related deaths in stem cell transplant patients, and may be amenable to correction with adoptive immunotherapy providing T lymphocytes specific for A. fumigatus. However, a clinically usable source of antigen and a reliable procedure for the generation of large numbers of Aspergillus-specific T lymphocytes to clinical-grade standards is not available.
  
  
  METHODS
  An environmental strain of A. fumigatus (WMAfES) was isolated and cultured using materials and reagents suitable for clinical manufacture. Water-soluble lysate from germinated conidia of WMAfES was used as the antigen source. Peripheral blood mononuclear cells were stimulated with antigen-pulsed autologous dendritic cells on days 0 and 7. Cells were expanded with a cocktail of interleukin (IL)-2, IL-7 and IL-15 from days 7 to 21.
  
  
  RESULTS
  We obtained a mean 32.8-fold increase in cell numbers over 21 days of culture (n = 8). Resultant cultures were predominantly effector and central memory CD4(+) T cells, which produced T-helper (h)1 and Th17 cytokines when restimulated with A. fumigatus antigen derived from environmental or clinically isolated A. fumigatus. Cultured cells exhibited a high level of specific expansion and chemokine production when restimulated. Moreover, cultured cells cross-reacted with antigens from other fungi, including Penicillium, Candida albicans and other non-fumigatus Aspergillus species.
  
  
  CONCLUSIONS
  We describe a simple, robust, reproducible and clinically applicable procedure using a clinically appropriate antigen preparation for the expansion of polyfunctional A. fumigatus-specific T cells from normal donors of varying HLA types.
authors.affiliation: Westmead Institute for Cancer Research, Westmead Millennium Institute and Faculty of Medicine, The University of Sydney, Westmead, New South Wales, Australia. shivashni.gaundar@sydney.edu.au
authors.forename: Shivashni S
authors.initials: SS
authors.lastname: Gaundar
authors.forename: Leighton
authors.initials: L
authors.lastname: Clancy
authors.forename: Emily
authors.initials: E
authors.lastname: Blyth
authors.forename: Wieland
authors.initials: W
authors.lastname: Meyer
authors.forename: David J
authors.initials: DJ
authors.lastname: Gottlieb
mesh_terms: D000946:Antigens, Fungal / Q000097:blood / Q000276:immunology; D001228:Aspergillosis / Q000276:immunology / Q000628:therapy; D001232:Aspergillus fumigatus / Q000254:growth & development / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes; D002478:Cells, Cultured; D018925:Chemokines / Q000276:immunology / Q000378:metabolism; D016207:Cytokines / Q000276:immunology / Q000378:metabolism; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology; D018380:Hematopoietic Stem Cell Transplantation / Q000009:adverse effects; D006801:Humans; D016219:Immunotherapy, Adoptive; D013172:Spores, Fungal / Q000276:immunology; D018417:Th1 Cells / Q000166:cytology / Q000276:immunology; D058504:Th17 Cells / Q000166:cytology / Q000276:immunology
chemical_list: D000946:Antigens, Fungal; D018925:Chemokines; D016207:Cytokines