title: Stimulation with lysates of Aspergillus terreus, Candida krusei and Rhizopus oryzae maximizes cross-reactivity of anti-fungal T cells.
abstract: OBJECTIVE
  Invasive fungal diseases caused by filamentous fungi and yeasts are significant causes of morbidity and mortality in immunosuppressed hematology patients. We previously published a method to expand Aspergillus fumigatus-specific T cells for clinical cell therapy. In the present study, we investigated expansion of T cells specific for other fungal pathogens and creation of a broadly reactive panfungal T-cell product.
  
  
  METHODS
  Fungal strains selected were those frequently observed in the clinical hematology setting and included Aspergillus, Candida, Fusarium, Rhizopus and Lomentospora/Scedosporium. Four T-cell cultures specific to each fungus were established. We selected lysates of Aspergillus terreus, Candida krusei and Rhizopus oryzae to expand panfungal T cells. Allelic restriction of anti-fungal activity was determined through the use of specific major histocompatibility complex class II-blocking antibodies.
  
  
  RESULTS
  Individual T-cell cultures specific to each fungus could be expanded in vitro, generating predominantly CD4(+) T cells of which 8% to 20% were fungus-specific. We successfully expanded panfungal T cells from the peripheral blood (n = 8) and granulocyte-colony-stimulating factor-primed stem cell products (n = 3) of normal donors by using a combination of lysates from Aspergillus terreus, Candida krusei and Rhizopus oryzae. Anti-fungal activity was mediated through human leukocyte antigen (HLA)-DR alleles and was maintained when antigen-presenting cells from partially HLA-DRB1-matched donors were used to stimulate T cells.
  
  
  CONCLUSIONS
  We demonstrate a method to manufacture panfungal T-cell products with specificity against a range of clinical fungal pathogens by use of the blood and stem cells of healthy donors as the starting material. The safety and efficacy of these products will need to be tested clinically.
authors.affiliation: Centre for Cancer Research, Westmead Millennium Institute for Medical Research, Sydney, Australia; Sydney Medical School, University of Sydney, Australia. Electronic address: shivashni.deo@sydney.edu.au.
authors.forename: Shivashni S
authors.initials: SS
authors.lastname: Deo
authors.affiliation: Centre for Cancer Research, Westmead Millennium Institute for Medical Research, Sydney, Australia.
authors.forename: Balaji
authors.initials: B
authors.lastname: Virassamy
authors.affiliation: Centre for Infectious Diseases and Microbiology, Westmead Hospital, Sydney, Australia.
authors.forename: Catriona
authors.initials: C
authors.lastname: Halliday
authors.affiliation: Sydney Cellular Therapies Laboratory, Westmead Hospital, Sydney, Australia.
authors.forename: Leighton
authors.initials: L
authors.lastname: Clancy
authors.affiliation: Sydney Medical School, University of Sydney, Australia; Centre for Infectious Diseases and Microbiology, Westmead Hospital, Sydney, Australia.
authors.forename: Sharon
authors.initials: S
authors.lastname: Chen
authors.affiliation: Sydney Medical School, University of Sydney, Australia; Centre for Infectious Diseases and Microbiology, Westmead Hospital, Sydney, Australia; Marie Bashir Institute for Infectious Diseases and Biosecurity, University of Sydney, Australia.
authors.forename: Wieland
authors.initials: W
authors.lastname: Meyer
authors.affiliation: Sydney Medical School, University of Sydney, Australia; Centre for Infectious Diseases and Microbiology, Westmead Hospital, Sydney, Australia; Marie Bashir Institute for Infectious Diseases and Biosecurity, University of Sydney, Australia.
authors.forename: Tania C
authors.initials: TC
authors.lastname: Sorrell
authors.affiliation: Centre for Cancer Research, Westmead Millennium Institute for Medical Research, Sydney, Australia; Sydney Medical School, University of Sydney, Australia; Sydney Cellular Therapies Laboratory, Westmead Hospital, Sydney, Australia; Blood and Marrow Transplant Unit, Department of Haematology, Westmead Hospital, Sydney, Australia.
authors.forename: David J
authors.initials: DJ
authors.lastname: Gottlieb
mesh_terms: D000938:Antigen-Presenting Cells / Q000276:immunology; D000946:Antigens, Fungal / Q000276:immunology; D001230:Aspergillus / Q000276:immunology*; D002175:Candida / Q000276:immunology*; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D003429:Cross Reactions / Q000276:immunology*; D016207:Cytokines / Q000096:biosynthesis; D000949:Histocompatibility Antigens Class II / Q000276:immunology; D006801:Humans; D025301:Hyphae / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology*; D018655:Lymphocyte Count; D010641:Phenotype; D012233:Rhizopus / Q000276:immunology*; D013045:Species Specificity; D013601:T-Lymphocytes / Q000276:immunology*; D014019:Tissue Donors
chemical_list: D000946:Antigens, Fungal; D016207:Cytokines; D000949:Histocompatibility Antigens Class II
keywords: Adoptive T-cell therapy; Aspergillus; Candida; Filamentous fungi; Fusarium; Hematopoietic stem cell transplantation; Invasive fungal disease; Scedosporium; Yeast; Zygomycetes