Optimized clinical-scale culture conditions for ex vivo selective depletion of host-reactive donor lymphocytes: A strategy for GvHD prophylaxis in allogeneic PBSC transplantation

Scott R. Solomon, T. Tran, C. S. Carter, S. Donnelly, N. Hensel, J. Schindler, E. Bahceci, V. Ghetie, J. Michálek, D. Mavroudis, E. J. Read, E. S. Vitetta, A. J. Barrett

Research output: Contribution to journalArticle

53 Scopus citations

Abstract

Background: Ex vivo selective depletion (SD) is a strategy to prevent GvHD, in which host-reactive donor lymphocytes are selectively eliminated from a PBSC allograft while conserving useful donor immune function. Prior to testing this strategy in patients, our goal was to develop a clinical-scale SD process, which involves co-culture of donor lymphocytes and irradiated recipient cells, followed by the addition of an immunotoxin (IT) directed against the α-chain of the IL-2 receptor (CD25), expressed on activated donor T cells. Methods: Stimulator cells were generated from immunomagnetically selected and expanded recipient T lymphocytes. Donor PBMCs from G-CSF-mobilized peripheral blood were co-cultured for 72 h with irradiated stimulator cells. Alloreactive T cells were targeted for elimination by the addition of the anti-CD25 IT, RFT5-SMPT-dgA, and the IT enhancer, NH4Cl. Results: Stimulator-cell selection/expansion yielded > 2×1010 highly enriched CD3+ cells (98.9 ± 2.2%). After SD, cell recovery was 68.5 ± 23.3% and viability was 84.6 ± 6.4%. This permitted a potential T-cell dose ≥ 1 × 108 CD3+ cells kg-1 to transplant recipients. Although SD donor lymphocytes retained little proliferative capacity against the original stimulator cells (2.6 ± 0.6%), responses were conserved against third party cells (107.6 ± 18.6%), the bacterial superantigen staphylococcus enterotoxin B (108.2 ± 4.2%), and CMV Ag (72.1 ± 3.8%). Discussion: We have demonstrated that ex vivo SD is feasible in clinical-scale culture conditions. The ability of this strategy to prevent GvHD is the subject of an ongoing clinical trial, in which the SD lymphocyte product is transplanted in conjunction with a T cell-depleted PBSC allograft.

Original languageEnglish (US)
Pages (from-to)395-406
Number of pages12
JournalCytotherapy
Volume4
Issue number5
DOIs
StatePublished - Jan 1 2002

Keywords

  • Allogeneic stem cell transplantation
  • Alloreactivity
  • Ex vivo cell culture
  • Expansion
  • Graft-versus-host disease
  • T-cell depletion

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Oncology
  • Genetics(clinical)
  • Cell Biology
  • Cancer Research
  • Transplantation

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    Solomon, S. R., Tran, T., Carter, C. S., Donnelly, S., Hensel, N., Schindler, J., Bahceci, E., Ghetie, V., Michálek, J., Mavroudis, D., Read, E. J., Vitetta, E. S., & Barrett, A. J. (2002). Optimized clinical-scale culture conditions for ex vivo selective depletion of host-reactive donor lymphocytes: A strategy for GvHD prophylaxis in allogeneic PBSC transplantation. Cytotherapy, 4(5), 395-406. https://doi.org/10.1080/146532402320775982