Clinical Treg cell applications for immunotherapy

  • Clinical-scale Treg cell depletion and enrichment
  • Convenient combination of CliniMACS® Platform with MACS® GMP Products
  • Customized strategies for manufacturing of Treg cell products
Regulatory T (Treg) cells belong to the CD4+ T cell subset. Their unique function, the suppression of immune responses, has made them an interesting target cell population for immunotherapy in a number of different indications in mice and human, including treatment and prevention of graft-versus-host disease (GvHD) after haploid stem cell transplantation (HSCT)1-4, suppression of solid organ rejection after transplantation5-9, and treatment of autoimmune diseases like type 1 diabetes10. To address the variety of Treg cell applications arising from different target indications, Miltenyi Biotec provides a broad range of tools and reagents facilitating the isolation and expansion of various Treg cell populations.
MACS Handbook:

Regulatory T cells (human)

Clinical-scale enrichment of Treg cells

Clinical-scale CD25+ cell separation from leukapheresis products using the enrichment protocol for cells expressing high levels of CD25
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Magnetic cell isolation or depletion of CD25+ Treg cells using the CliniMACS® System

Treg cells for clinical applications can be either enriched or depleted using the CliniMACS Cell Separation System in combination with the CliniMACS CD25 Reagent.

Cells expressing high levels of CD25 can be enriched with or without prior depletion of CD8+ or CD19+ cells.




Staining of Treg cells with MACS GMP Fluorescent Antibodies
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Flow sorting using MACS® GMP Antibodies

MACS GMP Fluorescent Antibodies have been developed for flow cytometric detection and subsequent flow sorting of highly pure Treg cell populations in clinical settings.

MACS GMP CD25-PE-Biotin and CliniMACS Anti-Biotin Reagent can be used for magnetic pre-enrichment of Treg cells from large volumes, followed by flow cytometric sorting for a defined and pure final product.

MACS Handbook:

Magnetic cell separation

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Clinical-scale Treg cell expansion

MACS GMP ExpAct Treg Kit

The MACS GMP ExpAct Treg Kit has been designed to efficiently expand Treg cells after a CliniMACS CD25 enrichment procedure while maintaining FoxP3 expression. It can be used in combination with TexMACS GMP Medium and MACS GMP Rapamycin, which inhibits the expansion of conventional T cells and thereby enhances the proportion of expanded Treg cells in culture.11,12

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Customer report
Clinical-grade separation and expansion of regulatory T cells for clinical studies on cell therapy in solid organ transplantation

Henrieta Fazekasova, Sarah Thirkell, Katie Lowe, Andrew Bushell, and Giovanna Lombardi, Immunoregulation laboratory, MRC Centre for Transplantation, King’s College London, Guy’s Hospital, London, UK

MACS Handbook:

Cell culture

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MACS GMP ExpAct Treg Kit (reference list)

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References

  1. Martelli, M. F. et al. (2014) HLA‑haploidentical transplantation with regulatory and conventional T‑cell adoptive immunotherapy prevents acute leukemia relapse. Blood 124(4): 638–644
  2. Martelli, M. F. et al. (2015) Next generation HLA-haploidentical HSCT. Bone Marrow Transplant. 50: 63–66.
  3. Theil, A. et al. (2015) Adoptive transfer of allogeneic regulatory T cells into patients with chronic graft‑versus‑host disease. Cytotherapy 17(4): 473–486.
  4. Hannon, M. et al. (2014) Infusion of clinical‑grade enriched regulatory T cells delays experimental xenogeneic graft‑versus‑host disease. Transfusion 54(2): 353–363.
  5. Nadig, S.N. et al. (2010) In vivo prevention of transplant arteriosclerosis by ex vivo-expanded human regulatory T cells. Nat. Med. 16: 809.
  6. Sagoo, P. et al. (2011) Human regulatory T cells with alloantigen specificity are more potent inhibitors of alloimmune skin graft damage than polyclonal regulatory T cells. Sci. Transl. Med. 3: 83ra42.
  7. Issa, F. et al. (2010) Ex vivo expanded human regulatory T cells prevent the rejection of skin allografts in a humanized mouse model. Transplantation 90: 1321.
  8. Wu, D.C. et al. (2013) Ex vivo expanded human regulatory T cells can prolong survival of a human islet allograft in a humanized mouse model. Transplantation 96: 707.
  9. Safina, N. et al. (2016) Successful expansion of functional and stable regulatory T cells for immunotherapy in liver transplantation. Oncotarget 7(7): 7563–7577.
  10. Bluestone J.A. (2015) Type 1 diabetes immunotherapy using polyclonal regulatory T cells. Sci. Transl. Med. 7, 315ra189.
  11. Putnam A. L. et al. (2013) Clinical grade manufacturing of human alloantigen-reactive regulatory T cells for use in transplantation. A. J. Transplant. 13: 3010–3020.
  12. Battaglia, M. et al. (2005) Rapamycin selectively expands CD4+CD25+FoxP3+ regulatory T cells. Blood 105: 4743–4748.

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