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Stimulation with lysates of Aspergillus terreus, Candida krusei and Rhizopus oryzae maximizes cross-reactivity of anti-fungal T cells

  • Shivashni S. Deo
    Correspondence
    Correspondence: Shivashni S. Deo, MD, Centre for Cancer Research, Westmead Millennium Institute, 176 Hawkesbury Road, Westmead, NSW 2145, Australia.
    Affiliations
    Centre for Cancer Research, Westmead Millennium Institute for Medical Research, Sydney, Australia

    Sydney Medical School, University of Sydney, Australia
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  • Balaji Virassamy
    Affiliations
    Centre for Cancer Research, Westmead Millennium Institute for Medical Research, Sydney, Australia
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  • Catriona Halliday
    Affiliations
    Centre for Infectious Diseases and Microbiology, Westmead Hospital, Sydney, Australia
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  • Leighton Clancy
    Affiliations
    Sydney Cellular Therapies Laboratory, Westmead Hospital, Sydney, Australia
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  • Sharon Chen
    Affiliations
    Sydney Medical School, University of Sydney, Australia

    Centre for Infectious Diseases and Microbiology, Westmead Hospital, Sydney, Australia
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  • Wieland Meyer
    Affiliations
    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
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  • Tania C. Sorrell
    Affiliations
    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
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  • David J. Gottlieb
    Affiliations
    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
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Published:November 06, 2015DOI:https://doi.org/10.1016/j.jcyt.2015.09.013

      Abstract

      Background aims. 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.

      Abbreviations:

      HSCT (hematopoietic stem cell transplantation), PBMC (peripheral blood mononuclear cells), GVHD (graft-versus host disease), HPC (hematopoietic progenitor cells), PBSC (peripheral blood stem cells), HLA (human leukocyte antigen)

      Key Words

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