Clinical-scale isolation of the total Aspergillus fumigatus-reactive T-helper cell repertoire for adoptive transfer

Petra Bacher, Andrea Jochheim-Richter, Nadine Mockel-Tenbrink, Olaf Kniemeyer, Eva Wingenfeld, Regina Alex, Alice Ortigao, Darja Karpova, Thomas Lehrnbecher, Andrew J. Ullmann, Axel Hamprecht, Oliver Cornely, Axel A. Brakhage, Mario Assenmacher, Halvard Bonig, Alexander Scheffold

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

Background aims: Evidence of the criticality of the adaptive immune response for controlling invasive aspergillosis has been provided. This observation is supported by the fact that invasive aspergillosis, a grave complication of allogeneic stem cell transplantation, occurs long after myeloid reconstitution in patients with low T-cell engraftment and/or on immunosuppressants. Adoptive T-cell transfer might be beneficial, but idiosyncrasies of Aspergillus fumigatus and the anti- Aspergillus immune response render established selection technologies ineffective. Methods: We developed a Good Manufacturing Practice (GMP)-compliant protocol for preparation of A. fumigatus-specific CD4+ cells by sequentially depleting regulatory and cytotoxic T cells, activating A. fumigatus-specific T-helper cells with GMP-grade A. fumigatus lysate, and immuno-magnetically isolating them via the transiently up-regulated activation marker, CD137. Results: In 13 full-scale runs, we demonstrate robustness and feasibility of the approach. From 2 × 109 peripheral blood mononuclear cells, we isolated 27 × 103-318 × 103Aspergillus-specific T-helper cells. Frequency among total T cells was increased, on average, by 200-fold. Specific studies indicate specificity and functionality: After non-specific in vitro expansion and re-stimulation with different antigens, we observed strong cytokine responses to A. fumigatus and some other fungi including Candida albicans, but none to unrelated antigens. Discussion: Our technology isolates naturally occurring Aspergillus-specific T-helper cells within 2 days of identifying the clinical indication. Rapid adoptive transfer of Aspergillus-specific T cells may be quite feasible; the clinical benefit remains to be demonstrated. A manufacturing license as an advanced-therapy medicinal product was received and a clinical trial in post-transplantation invasive aspergillosis patients approved. The product is dosed at 5 × 10E3/kg T cells (single intravenous injection), of which at least 10% must be A. fumigatus-specific.

Original languageEnglish
Pages (from-to)1396-1405
Number of pages10
JournalCytotherapy
Volume17
Issue number10
DOIs
StatePublished - Oct 1 2015

Keywords

  • Aspergillosis
  • Cell therapy
  • GMP-compliant protocol
  • Prospective isolation
  • T-helper cells

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