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Depletion of naive T cells using clinical grade magnetic CD45RA beads: a new approach for GVHD prophylaxis

Bone Marrow Transplantation volume 49, pages 138–144 (2014)Cite this article

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Abstract

Depletion of naive T cells from donor leukapheresis products (LPs) aims at the reduction of alloreactivity, while preserving memory T-cell reactivity (for example, to pathogens). This study established the immunomagnetic depletion procedure under clean room conditions using CD45RA beads and analyzed LPs of six donors for cell composition and functional immune responses. CD45RA depletion resulted in 3.4–4.7 log (median 4.4) reduction of CD45RA+ T cells, thereby eliminating naive and late effector T cells. B cells were also completely removed, whereas significant proportions of NK cells, monocytes and granulocytes persisted. CD45RA-depleted LPs contained effector and central memory CD4+ and CD8+ T cells that showed sustained IFN-γ secretion to CMV, EBV, Aspergillus and Candida Ags. Alloreactivity was measured in MLRs between donors with complete HLA-mismatch. Alloreactive CD8+ T cells were strongly reduced (median >1-log) upon CD45RA depletion, whereas alloreactive CD4+ T cells persisted in significant numbers. In conclusion, clinical grade depletion of CD45RA+ naive T cells from donor LPs is feasible and highly efficient. The depleted products show sustained CD4+ and CD8+ T-cell reactivity to pathogens and effectively reduced CD8-mediated alloreactivity. Prophylactic and preemptive infusions after allogeneic SCT may improve T-cell reconstitution and pathogen-specific immunosurveillance, along with lower risk of inducing GVHD.

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Acknowledgements

We thank E Schnürer for excellent technical assistance. We also thank the staffs of the good manufacturing practice cell production laboratory and Transfusion Center for performing CliniMACS and leukapheresis procedures, respectively. We are grateful to Dr O Kurzai (Hans-Knöll-Institute Jena, Germany) for providing Candida albicans germ tubes and to Dr A Scheffold (Miltenyi Biotec) for providing Aspergillus fumigatus lysate. This work was supported by grant KFO183-TP5 from the Deutsche Forschungsgemeinschaft to ED and WH, and by unrestricted provision of CD45RA depletion reagents from Miltenyi Biotec, Bergisch Gladbach, Germany.

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Author notes

  1. D Teschner and E Distler: These authors contributed equally to this work.

Authors and Affiliations

  1. Third Department of Medicine—Hematology, Oncology, Pneumology, University Medical Center of Johannes Gutenberg-University, Mainz, Germany

    D Teschner, E Distler, D Wehler, M Frey, M Theobald, S Thomas & W Herr

  2. Transfusion Center, University Medical Center of Johannes Gutenberg-University, Mainz, Germany

    D Marandiuc

  3. Miltenyi Biotec GmbH, Bergisch Gladbach, Germany

    K Langeveld

  4. Department of Internal Medicine III—Hematology and Oncology, University Medical Center of Regensburg, Regensburg, Germany

    W Herr

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  1. D Teschner
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  2. E Distler
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  3. D Wehler
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  4. M Frey
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  5. D Marandiuc
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  6. K Langeveld
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  7. M Theobald
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  8. S Thomas
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  9. W Herr
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Corresponding author

Correspondence to W Herr.

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Competing interests

Kirsten Langeveld is employee of the company Miltenyi Biotec, which provided the CD45RA depletion reagents free of charge. Miltenyi Biotec did not influence the design of the study. The company was also not involved in collection and analysis of data as well as the decision to publish the results. All authors including Kirsten Langeveld declare no conflict of interest.

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Teschner, D., Distler, E., Wehler, D. et al. Depletion of naive T cells using clinical grade magnetic CD45RA beads: a new approach for GVHD prophylaxis. Bone Marrow Transplant 49, 138–144 (2014). https://doi.org/10.1038/bmt.2013.114

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