Title |
Comparative Analysis of Clinical-Scale IFN-γ-Positive T-Cell Enrichment Using Partially and Fully Integrated Platforms
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Published in |
Frontiers in immunology, September 2016
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DOI | 10.3389/fimmu.2016.00393 |
Pubmed ID | |
Authors |
Christoph Priesner, Ruth Esser, Sabine Tischer, Michael Marburger, Krasimira Aleksandrova, Britta Maecker-Kolhoff, Hans-Gert Heuft, Lilia Goudeva, Rainer Blasczyk, Lubomir Arseniev, Ulrike Köhl, Britta Eiz-Vesper, Stephan Klöß |
Abstract |
The infusion of enriched CMV-specific donor T-cells appears to be a suitable alternative for the treatment of drug-resistant CMV reactivation or de novo infection after both solid organ and hematopoietic stem cell transplantation. Antiviral lymphocytes can be selected from apheresis products using the CliniMACS Cytokine-Capture-System(®) either with the well-established CliniMACS(®) Plus (Plus) device or with its more versatile successor CliniMACS Prodigy(®) (Prodigy). Manufacturing of CMV-specific T-cells was carried out with the Prodigy and Plus in parallel starting with 0.8-1 × 10(9) leukocytes collected by lymphapheresis (n = 3) and using the MACS GMP PepTivator(®) HCMVpp65 for antigenic restimulation. Target and non-target cells were quantified by a newly developed single-platform assessment and gating strategy using positive (CD3/CD4/CD8/CD45/IFN-γ), negative (CD14/CD19/CD56), and dead cell (7-AAD) discriminators. Both devices produced largely similar results for target cell viabilities: 37.2-52.2% (Prodigy) vs. 51.1-62.1% (Plus) CD45(+)/7-AAD(-) cells. Absolute numbers of isolated target cells were 0.1-3.8 × 10(6) viable IFN-γ(+) CD3(+) T-cells. The corresponding proportions of IFN-γ(+) CD3(+) T-cells ranged between 19.2 and 95.1% among total CD3(+) T-cells and represented recoveries of 41.9-87.6%. Within two parallel processes, predominantly IFN-γ(+) CD3(+)CD8(+) cytotoxic T-cells were enriched compared to one process that yielded a higher amount of IFN-γ(+) CD3(+)CD4(+) helper T lymphocytes. T-cell purity was higher for the Prodigies products that displayed a lower content of contaminating IFN-γ(-) T-cells (3.6-20.8%) compared to the Plus products (19.9-80.0%). The manufacturing process on the Prodigy saved both process and hands-on time due to its higher process integration and ability for unattended operation. Although the usage of both instruments yielded comparable results, the lower content of residual IFN-γ(-) T-cells in the target fractions produced with the Prodigy may allow for a higher dosage of CMV-specific donor T-cells without increasing the risk for graft-versus-host disease. |
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Switzerland | 1 | 100% |
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Members of the public | 1 | 100% |
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Unknown | 37 | 100% |
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Other | 4 | 11% |
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Student > Doctoral Student | 3 | 8% |
Student > Bachelor | 3 | 8% |
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Other | 5 | 14% |
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