The relative merits of cord blood as a cell source for autologous T regulatory cell therapy in type 1 diabetes.
Cord blood has been used as a cell source for therapeutic purposes in children with type 1 diabetes and other disorders. Here, we explore the benefits of cord blood as an autologous source of T regulatory cells for immune cell therapy in patients. CD4(+)CD25(+) T regulatory cells were isolated from cord blood and adult peripheral blood of healthy donors and compared during and after expansion in a 14-day protocol incorporating anti-CD3/anti-CD28 beads, and IL-2 with or without rapamycin. Cord blood T regulatory cells were largely naïve (89±7 vs. 31±10% in young adults, p<0.0001), and had higher expansion yields (median 5,968-fold) than adult T regulatory cells (median 516-fold, p=0.001) and adult naïve T regulatory cells (median 820-fold, p=0.003). Rapamycin reduced expansion yields, but was not necessary to obtain pure expanded cord blood T regulatory cells as judged by FOXP3 staining (94±3%), methylation status of FOXP3 (97%), and intracellular effector cytokine staining (< 6%). Expanded adult T regulatory cells were much less pure in the absence of rapamycin (72±19% FOXP3; 76% by methylation status, <13% INF-γ, <16% IL-4, <5% IL-17 positive), but purity was achieved by inclusion of rapamycin during expansion. Despite differences in purity, all preparations of expanded T regulatory from all sources were able to strongly suppress proliferation of T effector cells in vitro. Our findings suggest that cord blood is an excellent source of T regulatory cells for expansion and autologous cell therapy that may be considered as a strategy to prevent immune-mediated destruction of beta cells in type 1 diabetes.
DFG-Center for Regenerative Therapies, Technische Universität Dresden, Dresden, Germany.
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A
Theil
DFG-Center for Regenerative Therapies, Technische Universität Dresden, Dresden, Germany.
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C
Wilhelm
DFG-Center for Regenerative Therapies, Technische Universität Dresden, Dresden, Germany.
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Guhr
DFG-Center for Regenerative Therapies, Technische Universität Dresden, Dresden, Germany.
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J
Reinhardt
DFG-Center for Regenerative Therapies, Technische Universität Dresden, Dresden, Germany.
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Bonifacio
D000293:Adolescent; D000328:Adult; D015704:CD4 Antigens / Q000378:metabolism; D002452:Cell Count; D049109:Cell Proliferation; D002469:Cell Separation; D003922:Diabetes Mellitus, Type 1 / Q000628:therapy*; D005312:Fetal Blood / Q000166:cytology*; D006801:Humans; D053645:Interleukin-2 Receptor alpha Subunit / Q000378:metabolism; D010641:Phenotype; D050378:T-Lymphocytes, Regulatory / Q000166:cytology* / Q000276:immunology; D014182:Transplantation, Autologous; D055815:Young Adult
D015704:CD4 Antigens; D053645:Interleukin-2 Receptor alpha Subunit