An Innovative Standard Operation Procedure for Isolating GMP-Grade CD4+CD25+ T Cells from Non-Mobilized Leukapheresis
This SOP describes a closed system for isolating GMP-grade CD4+CD25+ T cells from non-mobilized leukapheresis collections (nMLCs), independent of a clean room in a certified GMP premises, by using CliniMACS format GMP grade reagents (CD25-labeled magnetic beads with/without pre-depletion of CD8+ T cells and CD19+ B cells), a GMP grade-A laminar hood and CliniMACS cell processing system.
Key wordsGMP CD25 Leukapheresis collection CliniMACS Magnetic beads
We would like to acknowledge the support of NHS Blood and Transplant, the National Institute for Health Biomedical Research Centre Program and the NHSBT and Department of Haematology Trust Funds. This report presents independent research commissioned by the National Institutes for Health Research (NIHR) under its Programme Grants for Applied Research Programme (Grant Reference Number RP-PG-0310-1003) (WZ, DJR, SMW). The views expressed in this publication are those of the authors and not necessarily those of the NHS, the NIHR, or the Department of Health.
The authors wish to thank all donors who participated in this study and the Therapeutic Apheresis Services Unit NHSBT Oxford for providing the leukapheresis collections.
- 5.Zhang W, Frith E, Belfield H, Smythe J, Clarke S, Watt SM, Danby R, Benjamin S, Peniket A, Roberts DJ (2015) An innovative method to generate a GMP-grade regulatory T cell product from non-mobilised leukapheresis donors independent of a clean room facility. CytotheropyGoogle Scholar
- 12.Trzonkowski P, Bieniaszewska M, Juścińska J, Dobyszuk A, Krzystyniak A, Marek N, Myśliwska J, Hellmann A (2009) First-in-man clinical results of the treatment of patients with graft versus host disease with human ex vivo expanded CD4+CD25+CD127- T regulatory cells. Clin Immunol 133(1):22–26CrossRefGoogle Scholar
- 13.Di Ianni M, Falzetti F, Carotti A, Terenzi A, Del Papa B, Perruccio K, Ruggeri L, Sportoletti P, Rosati E, Marconi P, Falini B, Reisner Y, Velardi A, Aversa F, Martelli MF (2011) Immunoselection and clinical use of T regulatory cells in HLA-haploidentical stem cell transplantation. Best Pract Res Clin Haematol 24(3):459–466CrossRefGoogle Scholar
- 15.Brunstein CG, Miller JS, Cao Q, McKenna DH, Hippen KL, Curtsinger J, Defor T, Levine BL, June CH, Rubinstein P, McGlave PB, Blazar BR, Wagner JE (2011) Infusion of ex vivo expanded T regulatory cells in adults transplanted with umbilical cord blood: safety profile and detection kinetics. Blood 117(3):1061–1070CrossRefGoogle Scholar
- 16.A G, Landau DA, Martin GH, Bonduelle O, Grinberg-Bleyer Y, Matheoud D, Grégoire S, Baillou C, Combadière B, Piaggio E, Cohen JL (2011) Immune reconstitution is preserved in hematopoietic stem cell transplantation coadministered with regulatory T cells for GVHD prevention. Blood 117(10):2975–2983CrossRefGoogle Scholar
- 17.Martelli MF, Di Ianni M, Ruggeri L, Falzetti F, Carotti A, Terenzi A, Pierini A, Massei MS, Amico L, Urbani E, Del Papa B, Zei T, Iacucci Ostini R, Cecchini D, Tognellini R, Reisner Y, Aversa F, Falini B, Velardi A (2014) HLA-haploidentical transplantation with regulatory and conventional T-cell adoptive immunotherapy prevents acute leukemia relapse. Blood 124(4):638–644CrossRefGoogle Scholar