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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References
- Bloodstream infections and invasive mycoses in children undergoing acute leukaemia treatment: a 13-year experience at a single Italian institution.Eur J Cancer. 2005; 41: 1439-1445
- The epidemiology of fungal infections in patients with hematologic malignancies: the SEIFEM-2004 study.Haematologica. 2006; 91: 1068-1075
- Epidemiology of paediatric invasive fungal infections and a case-control study of risk factors in acute leukaemia or post stem cell transplant.Br J Haematol. 2010; 149: 263-272
- Role of prospective screening of blood for invasive aspergillosis by polymerase chain reaction in febrile neutropenic recipients of haematopoietic stem cell transplants and patients with acute leukaemia.Br J Haematol. 2006; 132: 478-486
- Risk factors for the development of invasive fungal infections in allogeneic blood and marrow transplant recipients.Transpl Infect Dis. 2002; 4: 3-9
- Prospective surveillance for invasive fungal infections in hematopoietic stem cell transplant recipients, 2001–2006: overview of the Transplant-Associated Infection Surveillance Network (TRANSNET) Database.Clin Infect Dis. 2010; 50: 1091-1100
- Changing patterns and trends in systemic fungal infections.J Antimicrob Chemother. 2005; 56: i5-11
- Epidemiology and outcome of invasive fungal infection in adult hematopoietic stem cell transplant recipients: analysis of Multicenter Prospective Antifungal Therapy (PATH) Alliance registry.Clin Infect Dis. 2009; 48: 265
- Epidemiology and outcome of mould infections in hematopoietic stem cell transplant recipients.Clin Infect Dis. 2002; 34: 909-917
- Correlation between in-vitro susceptibility testing to itraconazole and in-vivo outcome of Aspergillus fumigatus infection.J Antimicrob Chemother. 1997; 40: 401-414
- The new fungal opportunists are coming.Clin Infect Dis. 1996; 22: S112-18
- Alteration of cell wall composition leads to amphotericin B resistance in Aspergillus flavus.Microbiol Immunol. 1999; 43: 1017-1025
- In vitro amphotericin B resistance in clinical isolates of Aspergillus terreus, with a head-to-head comparison to voriconazole.J Clin Microbiol. 1999; 37: 2343-2345
- Fungal infections in leukemia patients: how do we prevent and treat them?.Clin Infect Dis. 2010; 50: 405-415
- Epidemiology and outcomes of candidemia in 2019 patients: data from the prospective antifungal therapy alliance registry.Clin Infect Dis. 2009; 48: 1695-1703
- Retrospective study of candidemia in patients with hematological malignancies. Clinical features, risk factors and outcome of 76 episodes.Eur J Haematol. 1999; 63: 77-85
- Azole resistant Aspergillus fumigatus: an emerging problem.Med Mal Infect. 2013; 43: 139-145
- Interferon-gamma and tumor necrosis factor-alpha protect mice from invasive aspergillosis.J Infect Dis. 1995; 172: 1554-1560
- Tumor necrosis factor alpha enhances antifungal activities of polymorphonuclear and mononuclear phagocytes against Aspergillus fumigatus.Infect Immun. 1998; 66: 5999-6003
- Enumeration of functionally active anti-Aspergillus T-cells in human peripheral blood.J Immunol Methods. 2008; 335: 41-45
- Transferring functional immune responses to pathogens after haploidentical hematopoietic transplantation.Blood. 2005; 106: 4397-4406
- Robust polyfunctional T-helper 1 responses to multiple fungal antigens from a cell population generated using an environmental strain of Aspergillus fumigatus.Cytotherapy. 2012; 14: 1119-1130
- Identification of MHC class II restricted T-cell-mediated reactivity against MHC class I binding Mycobacterium tuberculosis peptides.Immunology. 2011; 132: 482-491
- Quantifying metabolic activity of filamentous fungi using a colorimetric XTT assay.Biotechnol Prog. 2008; 24: 780-783
- Generation of highly purified and functionally active human TH1 cells against Aspergillus fumigatus.Blood. 2006; 107: 2562-2569
- Human phagocytic cell responses to Scedosporium apiospermum (Pseudallescheria boydii): variable susceptibility to oxidative injury.Infect Immun. 2003; 71: 6472-6478
- Clinical-scale generation of multi-specific anti-fungal T cells targeting Candida, Aspergillus and mucormycetes.Cytotherapy. 2013; 15: 344-351
- Identification of immunogenic antigens from Aspergillus fumigatus by direct multiparameter characterization of specific conventional and regulatory CD4+ T cells.J Immunol. 2014; 193: 3332-3343
- Clinical-scale generation of human anti-Aspergillus T cells for adoptive immunotherapy.Bone Marrow Transplant. 2009; 43: 13-19
- Cytomegalovirus-specific cytotoxic T lymphocytes can be efficiently expanded from granulocyte colony-stimulating factor-mobilized hemopoietic progenitor cell products ex vivo and safely transferred to stem cell transplantation recipients to facilitate immune reconstitution.Biol Blood Marrow Transplant. 2013; 19: 725-734
- Multicenter study of banked third-party virus-specific T cells to treat severe viral infections after hematopoietic stem cell transplantation.Blood. 2013; 121: 5113-5123
- Cross-protective TH1 immunity against Aspergillus fumigatus and Candida albicans.Blood. 2011; 117: 5881-5891
- Generation of a multipathogen-specific T-cell product for adoptive immunotherapy based on activation-dependent expression of CD154.Blood. 2011; 118: 1121-1131
Article info
Publication history
Published online: November 06, 2015
Accepted:
September 23,
2015
Received:
July 28,
2015
Identification
Copyright
© 2015 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.
