Article Text

Download PDFPDF

P74. A Good Manufacturing Practice procedure to generate therapeutic numbers of highly pure anti-leukaemic virus-specific T-cells
  1. MM van Loenen1,
  2. R de Boer1,
  3. E van Liempt1,
  4. RS Hagedoorn1,
  5. P Meij2,
  6. I Jedema1,
  7. JHF Falkenburg1 and
  8. MHM Heemskerk1
  1. Aff1 grid.10419.3d0000000089452978HematologyLeiden University Medical Center Leiden the Netherlands
  2. Aff2 grid.10419.3d0000000089452978Clinical Pharmacy and ToxicologyLeiden University Medical Center Leiden the Netherlands

Statistics from

Request Permissions

If you wish to reuse any or all of this article please use the link below which will take you to the Copyright Clearance Center’s RightsLink service. You will be able to get a quick price and instant permission to reuse the content in many different ways.

Meeting abstracts


Recently, we have started a clinical trial to treat patients with high risk acute leukaemia with a donor-derived HA-1-TCR transduced virus-specific T-cell product as early as 8 weeks and 14 weeks after allogeneic stem cell transplantation (allo-SCT). Donor derived Cytomegalovirus (CMV)- and Epstein Bar virus (EBV)-specific T-cells will be isolated using Streptamer based CliniMACS selection, and will be subsequently transduced at day 2 with the well-characterized anti-leukaemic HA-1-TCR and infused 10-12 days later. Based on these well-defined specificities this T-cell product is predicted to result in a selective Graft versus Leukaemia (GvL) effect without Graft versus Host Disease (GvHD). Important study parameters are persistence of the T-cell product, feasibility of generation of HA-1-TCR transduced virus-specific T-cells, and the number of events of acute GvHD.

Material and methods

To obtain therapeutic cell numbers, one of the inclusion criteria is presence in donor peripheral blood of 1 or 2 virus-specific T-cell population with a frequency of ≥0.05% of T-cells. MHC-Streptamers will be used to isolate 1 or 2 virus-specific T-cell populations from donor leukocytes. MHC-Streptamer incubation will result in binding of the TCR of the virus-specific T-cells of interest to the specific peptide presented by the MHC molecule on the Streptamers. Next to allowing selection of T-cells of interest, this binding will also result in specific stimulation allowing subsequent transduction with the HA-1-TCR. The process of isolation of pure populations of virus-specific T-cells and transduction with good manufacturing practice (GMP)-grade retroviral supernatant encoding the HA-1-TCR has been validated with 4 large scale test procedures in the cleanroom. To pass the in process (IP) testing, T-cells needed to be ≥50% pure for the respective virus-specific tetramer directly after Streptamer isolation. In addition, after transduction and subsequent culturing T-cells need to be ≥60% antigen-specific, as measured with virus- and HA-1-tetramers. Moreover, transduction efficiency of ≥5% as measured with HA-1-tetramers is a prerequisite.


All HA-1-TCR td virus-specific T-cell products met the criteria for IP testing and quality control testing. They contained >90% antigen-specific T-cells and >10% HA-1 tetramer positive T-cells. Moreover, all HA-1-TCR td virus-specific T-cell products were highly reactive against HA-1-positive leukaemic cells.


Here, we present a GMP-grade procedure to generate in a short culture period of less than 2 weeks therapeutically relevant numbers of defined antigen-specific and highly anti-leukaemia reactive T-cells.