Stability of human rapamycin-expanded CD4+CD25+ T regulatory cells

TY - JOUR

T1 - Stability of human rapamycin-expanded CD4+CD25+ T regulatory cells

AU - Tresoldi, Eleonora

AU - Dell'Albani, Ilaria

AU - Stabilini, Angela

AU - Jofra, Tatiana

AU - Valle, Andrea

AU - Gagliani, Nicola

AU - Bondanza, Attilio

AU - Roncarolo, Maria Grazia

AU - Battaglia, Manuela

PY - 2011/9

Y1 - 2011/9

N2 - BACKGROUND: The clinical use of ex vivo-expanded T-regulatory cells for the treatment of T-cell-mediated diseases has gained increasing momentum. However, the recent demonstration that FOXP3(+) T-regulatory cells may contain interleukin-17-producing cells and that they can convert into effector cells once transferred in vivo raises significant doubts about their safety. We previously showed that rapamycin permits the ex vivo expansion of FOXP3(+) T-regulatory cells while impairing the proliferation of non-T-regulatory cells. Here we investigated the Th17-cell content and the in vivo stability of rapamycin-expanded T-regulatory cells as pertinent aspects of cell-based therapy.DESIGN AND METHODS: T-regulatory-enriched cells were isolated from healthy volunteers and were expanded ex vivo with rapamycin with a pre-clinical applicable protocol. T-regulatory cells cultured with and without rapamycin were compared for their regulatory activity, content of pro-inflammatory cells and stability.RESULTS: We found that CD4(+)CCR6(+)CD161(+) T cells (i.e., precursor/committed Th17 cells) contaminate the T-regulatory cells cultured ex vivo in the absence of rapamycin. In addition, Th17 cells do not expand when rapamycin-treated T-regulatory cells are exposed to a "Th17-favorable" environment. Rapamycin-expanded T-regulatory cells maintain their in vitro regulatory phenotype even after in vivo transfer into immunodeficient NOD-SCID mice despite being exposed to the irradiation-induced pro-inflammatory environment. Importantly, no additional rapamycin treatment, either in vitro or in vivo, is required to keep their phenotype fixed.CONCLUSIONS: These data demonstrate that rapamycin secures ex vivo-expanded human T-regulatory cells and provide additional justification for their clinical use in future cell therapy-based trials.

AB - BACKGROUND: The clinical use of ex vivo-expanded T-regulatory cells for the treatment of T-cell-mediated diseases has gained increasing momentum. However, the recent demonstration that FOXP3(+) T-regulatory cells may contain interleukin-17-producing cells and that they can convert into effector cells once transferred in vivo raises significant doubts about their safety. We previously showed that rapamycin permits the ex vivo expansion of FOXP3(+) T-regulatory cells while impairing the proliferation of non-T-regulatory cells. Here we investigated the Th17-cell content and the in vivo stability of rapamycin-expanded T-regulatory cells as pertinent aspects of cell-based therapy.DESIGN AND METHODS: T-regulatory-enriched cells were isolated from healthy volunteers and were expanded ex vivo with rapamycin with a pre-clinical applicable protocol. T-regulatory cells cultured with and without rapamycin were compared for their regulatory activity, content of pro-inflammatory cells and stability.RESULTS: We found that CD4(+)CCR6(+)CD161(+) T cells (i.e., precursor/committed Th17 cells) contaminate the T-regulatory cells cultured ex vivo in the absence of rapamycin. In addition, Th17 cells do not expand when rapamycin-treated T-regulatory cells are exposed to a "Th17-favorable" environment. Rapamycin-expanded T-regulatory cells maintain their in vitro regulatory phenotype even after in vivo transfer into immunodeficient NOD-SCID mice despite being exposed to the irradiation-induced pro-inflammatory environment. Importantly, no additional rapamycin treatment, either in vitro or in vivo, is required to keep their phenotype fixed.CONCLUSIONS: These data demonstrate that rapamycin secures ex vivo-expanded human T-regulatory cells and provide additional justification for their clinical use in future cell therapy-based trials.

KW - Animals

KW - CD4 Antigens

KW - Cell Proliferation

KW - Cells, Cultured

KW - Cytokines

KW - Female

KW - Forkhead Transcription Factors

KW - Humans

KW - Immunophenotyping

KW - Interleukin-2 Receptor alpha Subunit

KW - Leukocytes, Mononuclear

KW - Mice

KW - Mice, Inbred NOD

KW - Mice, SCID

KW - Sirolimus

KW - T-Lymphocytes, Regulatory

KW - Th17 Cells

KW - Journal Article

U2 - 10.3324/haematol.2011.041483

DO - 10.3324/haematol.2011.041483

M3 - SCORING: Journal article

C2 - 21565906

VL - 96

SP - 1357

EP - 1365

JO - HAEMATOLOGICA

JF - HAEMATOLOGICA

SN - 0390-6078

IS - 9

ER -