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Cell Procurement

Clinical-scale generation of human anti-Aspergillus T cells for adoptive immunotherapy

Bone Marrow Transplantation volume 43, pages 13–19 (2009)Cite this article

Abstract

Invasive aspergillosis is a major cause of morbidity and mortality in patients undergoing allogeneic hematopoietic SCT. There is a growing body of evidence that T cells are important in the host defense against Aspergillus, and adoptively transferred anti-Aspergillus T-helper 1 (TH) 1 cells might reduce infectious mortality in hematopoietic transplant recipients. Here we present for the first time a simple and rapid method for the clinical-scale generation of functionally active anti-Aspergillus T cells according to good manufacturing practice conditions. A total of 1.1 × 109 WBCs derived from a leukapheresis product were incubated with Aspergillus antigens. Stimulated cells were selected by means of the IFN-γ secretion assay and expanded. In three independent experiments, a median number of 2 × 107 CD3+CD4+cells (range, 0.9–3.2 × 107) were obtained within 13 days. The cultured CD3+CD4+ cells exhibited almost exclusively a memory activated T-helper cell phenotype. Upon restimulation, the generated T cells produced IFN-γ, but no IL-4 or IL-10, indicating a TH1-cell population. Additionally, the cells proliferated upon restimulation and showed reduced alloreactivity compared to unselected CD4+ cells. This method of generating is suitable for future prospective trials designed to evaluate the effect of adoptive immunotherapy in hematopoietic transplant recipients with invasive aspergillosis.

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Acknowledgements

We thank Carla Fadler, Nicola Schuely and Frauke Roeger for their technical assistance, and Klaus-Peter Hunfeld for microbiological testing. We also thank Georg Rauser for helpful discussions. This study was supported by the Deutsche Leukämie-Forschungshilfe (DLFH).

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Authors and Affiliations

  1. Pediatric Hematology and Oncology, Children's Hospital III, Johann Wolfgang Goethe University, Frankfurt, Germany

    L Tramsen, U Koehl, R Quaritsch, O Beck, T Klingebiel & T Lehrnbecher

  2. Red Cross Blood Donor Service Baden-Wuerttemberg-Hessen, Institute for Transfusion Medicine and Immunohematology, Johann Wolfgang Goethe University, Frankfurt, Germany

    T Tonn & E Seifried

  3. Insitut Pasteur, Aspergillus Unit, Paris, France

    J-P Latgé

  4. Clinics of Pediatric Oncology, Hematology and Immunology, University Hospital of Düsseldorf, Düsseldorf, Germany

    F R Schuster & A Borkhardt

  5. Department of Hematology, Oncology and Transfusion Medicine, Charité-Campus Benjamin Franklin, Berlin, Germany

    L Uharek

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  1. L Tramsen
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  2. U Koehl
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  3. T Tonn
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Correspondence to T Lehrnbecher.

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Tramsen, L., Koehl, U., Tonn, T. et al. Clinical-scale generation of human anti-Aspergillus T cells for adoptive immunotherapy. Bone Marrow Transplant 43, 13–19 (2009). https://doi.org/10.1038/bmt.2008.271

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