P74. A Good Manufacturing Practice procedure to generate therapeutic numbers of highly pure...
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POSTER PRESENTATION Open Access P74. A Good Manufacturing Practice procedure to generate therapeutic numbers of highly pure anti-leukaemic virus-specific T-cells MM van Loenen 1* , R de Boer 1 , E van Liempt 1 , RS Hagedoorn 1 , P Meij 2 , I Jedema 1 , JHF Falkenburg 1 , MHM Heemskerk 1 From 1st Immunotherapy of Cancer Conference (ITOC1) Munich, Germany. 12-14 March 2014 Background 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 Cytomegalo- virus (CMV)- and Epstein Bar virus (EBV)-specific T-cells will be isolated using Streptamer based Clini- MACS 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 iso- late 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 inter- est to the specific peptide presented by the MHC mole- cule 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 superna- tant 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. Results 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. Conclusions Here, we present a GMP-grade procedure to generate in a short culture period of less than 2 weeks therapeuti- cally relevant numbers of defined antigen-specific and highly anti-leukaemia reactive T-cells. Authors’ details 1 Leiden University Medical Center, Hematology, Leiden, the Netherlands. 2 Leiden University Medical Center, Clinical Pharmacy and Toxicology, Leiden, the Netherlands. 1 Leiden University Medical Center, Hematology, Leiden, the Netherlands Full list of author information is available at the end of the article van Loenen et al. Journal for ImmunoTherapy of Cancer 2014, 2(Suppl 2):P48 http://www.immunotherapyofcancer.org/content/2/S2/P48 © 2014 van Loenen et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Transcript of P74. A Good Manufacturing Practice procedure to generate therapeutic numbers of highly pure...
POSTER PRESENTATION Open Access
P74. A Good Manufacturing Practice procedure togenerate therapeutic numbers of highly pureanti-leukaemic virus-specific T-cellsMM van Loenen1*, R de Boer1, E van Liempt1, RS Hagedoorn1, P Meij2, I Jedema1, JHF Falkenburg1,MHM Heemskerk1
From 1st Immunotherapy of Cancer Conference (ITOC1)Munich, Germany. 12-14 March 2014
BackgroundRecently, we have started a clinical trial to treat patientswith high risk acute leukaemia with a donor-derivedHA-1-TCR transduced virus-specific T-cell product asearly as 8 weeks and 14 weeks after allogeneic stem celltransplantation (allo-SCT). Donor derived Cytomegalo-virus (CMV)- and Epstein Bar virus (EBV)-specificT-cells will be isolated using Streptamer based Clini-MACS selection, and will be subsequently transduced atday 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 toresult in a selective Graft versus Leukaemia (GvL) effectwithout Graft versus Host Disease (GvHD). Importantstudy 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 methodsTo obtain therapeutic cell numbers, one of the inclusioncriteria is presence in donor peripheral blood of 1 or 2virus-specific T-cell population with a frequency of≥0.05% of T-cells. MHC-Streptamers will be used to iso-late 1 or 2 virus-specific T-cell populations from donorleukocytes. MHC-Streptamer incubation will result inbinding of the TCR of the virus-specific T-cells of inter-est to the specific peptide presented by the MHC mole-cule on the Streptamers. Next to allowing selection ofT-cells of interest, this binding will also result in specificstimulation allowing subsequent transduction with the
HA-1-TCR. The process of isolation of pure populationsof virus-specific T-cells and transduction with goodmanufacturing practice (GMP)-grade retroviral superna-tant encoding the HA-1-TCR has been validated with 4large scale test procedures in the cleanroom. To pass thein process (IP) testing, T-cells needed to be ≥50% purefor the respective virus-specific tetramer directly afterStreptamer isolation. In addition, after transduction andsubsequent culturing T-cells need to be ≥60% antigen-specific, as measured with virus- and HA-1-tetramers.Moreover, transduction efficiency of ≥5% as measuredwith HA-1-tetramers is a prerequisite.
ResultsAll HA-1-TCR td virus-specific T-cell products met thecriteria for IP testing and quality control testing. Theycontained >90% antigen-specific T-cells and >10% HA-1tetramer positive T-cells. Moreover, all HA-1-TCR tdvirus-specific T-cell products were highly reactiveagainst HA-1-positive leukaemic cells.
ConclusionsHere, we present a GMP-grade procedure to generate ina short culture period of less than 2 weeks therapeuti-cally relevant numbers of defined antigen-specific andhighly anti-leukaemia reactive T-cells.
Authors’ details1Leiden University Medical Center, Hematology, Leiden, the Netherlands.2Leiden University Medical Center, Clinical Pharmacy and Toxicology, Leiden,the Netherlands.
1Leiden University Medical Center, Hematology, Leiden, the NetherlandsFull list of author information is available at the end of the article
van Loenen et al. Journal for ImmunoTherapy of Cancer 2014, 2(Suppl 2):P48http://www.immunotherapyofcancer.org/content/2/S2/P48
© 2014 van Loenen et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the CreativeCommons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, andreproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver(http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
Published: 12 March 2014
doi:10.1186/2051-1426-2-S2-P48Cite this article as: van Loenen et al.: P74. A Good ManufacturingPractice procedure to generate therapeutic numbers of highly pureanti-leukaemic virus-specific T-cells. Journal for ImmunoTherapy of Cancer2014 2(Suppl 2):P48.
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van Loenen et al. Journal for ImmunoTherapy of Cancer 2014, 2(Suppl 2):P48http://www.immunotherapyofcancer.org/content/2/S2/P48
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