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

« Previous Next »Biology of Blood and Marrow Transplantation
Volume 12, Issue 3 , Pages 267-274, March 2006

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

Petra Hoffmann
- Department of Hematology and Oncology, University Hospital Regensburg, Regensburg, Germany
- Institute of Immunology, University Regensburg, Regensburg, Germany
Tina J. Boeld
- Department of Hematology and Oncology, University Hospital Regensburg, Regensburg, Germany
Ruediger Eder
- Department of Hematology and Oncology, University Hospital Regensburg, Regensburg, Germany
Julia Albrecht
- Department of Hematology and Oncology, University Hospital Regensburg, Regensburg, Germany
Kristina Doser
- Department of Hematology and Oncology, University Hospital Regensburg, Regensburg, Germany
Biserka Piseshka
- Department of Hematology and Oncology, University Hospital Regensburg, Regensburg, Germany
Ashraf Dada
- Institute for Clinical Chemistry and Laboratory Medicine, University Hospital Regensburg, Regensburg, Germany
Claudia Niemand
- Miltenyi Biotec, Bergisch Gladbach, Germany
Mario Assenmacher
- Miltenyi Biotec, Bergisch Gladbach, Germany
Evelyn Orsó
- Institute for Clinical Chemistry and Laboratory Medicine, University Hospital Regensburg, Regensburg, Germany
Reinhard Andreesen
- Department of Hematology and Oncology, University Hospital Regensburg, Regensburg, Germany
Ernst Holler
- Department of Hematology and Oncology, University Hospital Regensburg, Regensburg, Germany
Matthias Edinger
- Department of Hematology and Oncology, University Hospital Regensburg, Regensburg, Germany
- Correspondence and reprint requests: Matthias Edinger, MD, Department of Hematology and Oncology, University Hospital Regensburg, Franz-Josef-Strauss-Allee 11, 93053 Regensburg, Germany

Received 7 December 2005; accepted 9 January 2006.

Abstract

The adoptive transfer of donor CD4⁺CD25⁺ regulatory T cells has been shown to protect from lethal graft-versus-host disease after allogeneic bone marrow transplantation in murine disease models. Efficient isolation strategies that comply with good manufacturing practice (GMP) guidelines are prerequisites for the clinical application of human CD4⁺CD25⁺ regulatory T cells. Here we describe the isolation of CD4⁺CD25⁺ T cells with regulatory function from standard leukapheresis products by using a 2-step magnetic cell-separation protocol performed under GMP conditions. The generated cell products contained on average 49.5% CD4⁺CD25^high T cells that phenotypically and functionally represented natural CD4⁺CD25⁺ regulatory T cells and showed a suppressive activity comparable to that of CD4⁺CD25⁺ regulatory T-cell preparations purified by non–GMP-approved fluorescence-activated cell sorting.