Isolation of CD4+CD25+ Regulatory T Cells for Clinical Trials

Hoffmann, Petra; Boeld, Tina J.; Eder, Ruediger; Albrecht, Julia; Doser, Kristina; Piseshka, Biserka; Dada, Ashraf; Niemand, Claudia; Assenmacher, Mario; Orsó, Evelyn; Andreesen, Reinhard; Holler, Ernst; Edinger, Matthias

doi:10.1016/j.bbmt.2006.01.005

Cited by 121 publications

(86 citation statements)

References 24 publications

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“…44 Adoptive transfer of regulatory T cells is also being evaluated as a therapy for GvHD. 51 Numbers of regulatory CD4 T cells were, however, not deficient in our GvHD patients and were actually increased in those with the chronic form of disease; furthermore, these cells exhibited suppressive activity in vitro. Additional studies are, therefore, required to understand why chronic GvHD pathology persists.…”

Section: Discussionmentioning

confidence: 54%

Imbalance of effector and regulatory CD4 T cells is associated with graft-versus-host disease after hematopoietic stem cell transplantation using a reduced intensity conditioning regimen and alemtuzumab

Matthews¹,

Lim²,

Afzali³

et al. 2009

Haematologica

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Section: Discussionmentioning

confidence: 54%

Imbalance of effector and regulatory CD4 T cells is associated with graft-versus-host disease after hematopoietic stem cell transplantation using a reduced intensity conditioning regimen and alemtuzumab

Matthews¹,

Lim²,

Afzali³

et al. 2009

Haematologica

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“…CD4 + CD25 high Treg cells currently evaluated in clinical trials [31,32] are cells isolated by the CliniMACS system, resulting in a FOXP3 + cell purity of just 40-70%. However, as FOXP3 is also expressed on activated human T cells and on unstable Tregcell populations [33][34][35], Treg-cell purity should in addition be evaluated by assessment of the methylation of the TSDR region, which is highly specific for stable Treg cells [36,37].…”

Section: Discussionmentioning

confidence: 99%

Isolation of human antigen‐specific regulatory T cells with high suppressive function

et al. 2014

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Adoptive transfer of regulatory T (Treg) cells could be an alternative to chronic immunosuppression for prevention of allogeneic graft rejection. While polyspecific Treg cells can prevent immune responses under lymphopenic conditions, Ag-specific Treg cells are needed to treat autoimmunity and graft rejection. Yet, reliable markers for Ag-specific Treg cells are missing. We report that latency-associated peptide (LAP) and glycoprotein A repetitions predominant (GARP) can identify human Ag-specific Treg cells. In addition,we show that the depletion of CD154 + cells from LAP + or GARP + Treg cells increases the Treg-cell purity to over 90%, as assessed by epigenetic analysis. These Ag-specific Treg cells can be isolated magnetically and might contribute to the development of GMP-based protocols. In addition, Ag-specific Treg cells are functionally far superior to CD4 + CD25 high or CD4 + CD25 high CD127 low Treg cells in vitro and in preventing strong alloreactions in humanized mice. They could, therefore, have a high therapeutic potential for the control of alloimmune, autoimmune, and allergic immune responses in patients.Keywords: Antigen-specific r CD154 r GARP r LAP r Regulatory T (Treg) cells Additional supporting information may be found in the online version of this article at the publisher's web-site IntroductionIn organ transplantation, acute rejection can be prevented by a combination of nonspecific immunosuppressive drugs and immunomodulatory induction therapies, yet chronic rejection and chronic graft dysfunction remain a clinical problem [1]. In addition, chronic nonspecific immunosuppression causes renal dysfunction and predisposes patients to infections and increased Correspondence: Dr. Elmar Jaeckel e-mail: Jaeckel.Elmar@mh-hannover.de risk of malignancies [1]. Innate and adaptive immune responses can be controlled by Treg cells, thereby offering new therapeutic options after organ transplantation [2][3][4][5][6]. CD4 + CD25 high FOXP3 + Treg cells have been shown to be of major importance in allospecific tolerance in animal models and to influence donor-specific immune responses in transplant patients [7][8][9][10]. After allogeneic hematopoietic stem cell transplantation, polyspecific Treg cells can prevent GVH disease without influencing * These authors contributed equally to this work.C 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.eji-journal.eu Eur. J. Immunol. 2014. 44: 2592-2602 Cellular immune response 2593 graft-versus-leukemia responses [11,12]. Therefore, polyspecific CD4 + CD25 high cells are currently being evaluated after stem cell transplantation [13]. However, under nonlymphopenic conditions as seen in patients with autoimmune diseases or in patients after organ transplantation, polyspecific Treg cells have so far been largely ineffective in controlling immune responses [5,14,15] [24], and CD137 (4-1BB) [25] were described as Treg-cell activation markers after polyspecific Treg-cell activation.In this manuscript, we demonstrated for the first time that LAP and GARP can be use...

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“…5 Protocols for the expansion of human FOXP3 + Treg cells performed under conditions of good manufacturing practice (GMP) have been shown to be feasible. 3,[6][7][8] Importantly, these culture conditions are also highly advantageous for the expansion of effector CD25 + T cells, which may contaminate the starting peripheral FOXP3 + Treg cells. This obstacle can be bypassed by: (i) the use of cord blood Treg cells, 4 (ii) isolation of very pure peripheral Treg cells through flow-based cell sorters 3,9,10 or (iii) the addition of rapamycin to the culture, which preferentially inhibits proliferation of effector T cells while sparing Treg cells.…”

Section: Introductionmentioning

confidence: 99%

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

Tresoldi¹,

Dell’Albani²,

Stabilini³

et al. 2011

Haematologica

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The online version of this article has a Supplementary Appendix. BackgroundThe 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 + Tregulatory 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 MethodsT-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. ResultsWe found that CD4+ 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. ConclusionsThese 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.Key words: T-regulatory cells, ex vivo expansion, rapamycin, cell therapy, T-regulatory-cell stability. + T regulatory cells. Haematologica 2011;96(9):1357-1365

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Isolation of CD4+CD25+ Regulatory T Cells for Clinical Trials

Cited by 121 publications

References 24 publications

Imbalance of effector and regulatory CD4 T cells is associated with graft-versus-host disease after hematopoietic stem cell transplantation using a reduced intensity conditioning regimen and alemtuzumab

Imbalance of effector and regulatory CD4 T cells is associated with graft-versus-host disease after hematopoietic stem cell transplantation using a reduced intensity conditioning regimen and alemtuzumab

Isolation of human antigen‐specific regulatory T cells with high suppressive function

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

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