Clinical-scale generation of multi-specific anti-fungal T cells targeting Candida, Aspergillus and mucormycetes

Cytotherapy. 2013 Mar;15(3):344-51. doi: 10.1016/j.jcyt.2012.11.014.

Abstract

Background aims: 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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aspergillus / immunology
  • Aspergillus / isolation & purification*
  • Aspergillus / pathogenicity
  • Candida / immunology
  • Candida / isolation & purification*
  • Candida / pathogenicity
  • Female
  • Flow Cytometry
  • Hematopoietic Stem Cell Transplantation / adverse effects
  • Hematopoietic Stem Cell Transplantation / mortality
  • Humans
  • Interferon-gamma / immunology*
  • Interferon-gamma / metabolism
  • Interferon-gamma Release Tests
  • Leukocytes, Mononuclear / cytology
  • Leukocytes, Mononuclear / immunology
  • Male
  • Middle Aged
  • Rhizopus / immunology
  • Rhizopus / isolation & purification*
  • Rhizopus / pathogenicity
  • T-Lymphocytes / cytology
  • T-Lymphocytes / immunology*

Substances

  • Interferon-gamma