Clinical Grade Treg: GMP Isolation, Improvement of Purity by CD127pos Depletion, Treg Expansion, and Treg Cryopreservation

• DOI: 10.1371/journal.pone.0003161
• Publication Date: 2008-09-05T15:24:35Z
• Authors: Jorieke H. Peters; Frank W. Preijers; Rob Woestenenk; Luuk B. Hilbrands; Hans J. P. M. Koenen; Irma Joosten
• Keywords: Interleukin-7 receptor; Regulatory T cell

Treg based immunotherapy is of great interest to facilitate tolerance in autoimmunity and transplantation. For clinical trials, it essential have a grade isolation protocol accordance with Good Manufacturing Practice (GMP) guidelines. To obtain sufficient for immunotherapy, subsequent ex vivo expansion might be needed.Treg were isolated from leukapheresis products by CliniMACS GMP strategies, using anti-CD25, anti-CD8 anti-CD19 coated microbeads. procedures led 40-60% pure CD4(pos)CD25(high)FoxP3(pos) populations that anergic had moderate suppressive activity. Such could expanded maintenance function. Alloantigen stimulated caused an enrichment alloantigen-specific Treg. Depletion unwanted CD19(pos) cells during proved necessary prevent B-cell outgrowth expansion. CD4(pos)CD127(pos) conventional T the major contaminating cell type populations. CD127(pos) improved purity approximately 90%. Expanded CD127(neg) showed very potent capacity high FoxP3 expression. Furthermore, our data show cryopreservation feasible, but activation after thawing restore potential.The feasibility therapy widely accepted, provided tailor-made handling are available. We here provide further support this approach showing can reached, cryopreserved successfully.