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Review
. 2019 Jul;197(1):52-63.
doi: 10.1111/cei.13297. Epub 2019 Apr 15.

Methods to manufacture regulatory T cells for cell therapy

K N MacDonald  1   2   3 , J M Piret  3   4 , M K Levings  1   2   5
Affiliations
Review

Methods to manufacture regulatory T cells for cell therapy

K N MacDonald et al. Clin Exp Immunol. 2019 Jul.

Abstract

Regulatory T cell (Treg ) therapy has shown promise in early clinical trials for treating graft-versus-host disease, transplant rejection and autoimmune disorders. A challenge has been to isolate sufficiently pure Tregs and expand them to a clinical dose. However, there has been considerable progress in the development and optimization of these methods, resulting in a variety of manufacturing protocols being tested in clinical trials. In this review, we summarize methods that have been used to manufacture Tregs for clinical trials, including the choice of cell source and protocols for cell isolation and expansion. We also discuss alternative culture or genome editing methods for modulating Treg specificity, function or stability that could be applied to future clinical manufacturing protocols to increase the efficacy of Treg therapy.

Keywords: autoimmunity; regulatory T cells; tolerance/suppression/anergy; transplantation.

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Figures

Figure 1
Figure 1
Overview of regulatory T cell (Treg) manufacturing protocols. Tregs are isolated from peripheral blood, cord blood or thymus tissue by magnetic cell separation or flow cytometric sorting. Isolated Tregs are then expanded using anti‐CD3/CD28 or artificial antigen‐presenting cells (K562 64/86 aAPCs), interleukin (IL)‐2 and in some cases rapamycin. After expansion, Tregs are harvested and directly administered to the patient or cryopreserved for future administration.
Figure 2
Figure 2
Good manufacturing practice (GMP) expansion protocols for polyclonal regulatory T cells (Tregs). In reported clinical protocols, Tregs are expanded for 7–36 days using anti‐CD3/CD28 beads or artificial antigen‐presenting cells (K562 64/86 aAPCs). During the expansion, medium is supplemented with a range of interleukin (IL)‐2 concentrations (200–1000 IU/ml) and in some cases rapamycin.
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