Infusion of clinical-grade enriched regulatory T cells delays experimental xenogeneic graft-versus-host disease

Authors

  • Muriel Hannon,

    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
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  • Chantal Lechanteur,

    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
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  • Sophie Lucas,

    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
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  • Joan Somja,

    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
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  • Laurence Seidel,

    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
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  • Ludovic Belle,

    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
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  • France Bruck,

    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
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  • Etienne Baudoux,

    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
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  • Olivier Giet,

    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
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  • Anne-Marie Chantillon,

    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
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  • Philippe Delvenne,

    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
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  • Pierre Drion,

    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
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  • Yves Beguin,

    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
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  • Stéphanie Humblet-Baron,

    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
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    • FB and SHB are co-senior authors.
  • Frédéric Baron

    Corresponding author
    1. Groupe Interdisciplinaire de Génoprotéomique Appliquée (GIGA)-I3, Laboratory of Cell and Genetic Therapy, Department of Pathology, Department of Statistics, GIGA-R, Department of Medicine, Division of Hematology, University of Liège, Liège, Belgium
    2. de Duve Institute, Université Catholique de Louvain, Brussels, Belgium
    3. Red Cross Transfusion Center of Liege, Liège, Belgium
    4. Autoimmune Genetics Laboratory, University of Leuven, Leuven, Belgium
    • Address reprint requests to: Frédéric Baron, Department of Hematology, University of Liège, CHU Sart-Tilman, 4000 Liège, Belgium; e-mail: f.baron@ulg.ac.be.

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    • FB and SHB are co-senior authors.

  • This study was supported by funds from the National Fund for Scientific Research (FNRS), the Leon Fredericq fund and Anti-Cancer Center at the University of Liège, and the Belgian Federation Against Cancer. MH and LB are Télévie Research Assistant, SHB is postdoctoral researcher, and SL and FB are senior research associates of the National Fund for Scientific Research (FNRS) Belgium.

Abstract

Background

We investigated the ability of clinical-grade enriched human regulatory T cells (Treg) to attenuate experimental xenogeneic graft-versus-host disease (GVHD) induced by peripheral blood mononuclear cells (PBMNCs; autologous to Treg) infusion in NSG mice, as well as verified their inability to induce xenogeneic GVHD when infused alone.

Study Design and Methods

Human Treg were isolated from peripheral blood apheresis products with a cell separation system (CliniMACS, Miltenyi Biotec GmbH) using a two-step procedure (simultaneous CD8 and CD19 depletion followed by CD25-positive selection) in six independent experiments with six different healthy volunteer donors. Sublethally (2.5 Gy) irradiated NSG mice were given 2 × 106 cytapheresis (PBMNC) product cells intravenously (IV) without (PBMNC group) or with 1 × 106 Treg (PBMNC + Treg group), while other NSG mice received 2 × 106 enriched Treg alone (also in IV; Treg group).

Results

The first five procedures were successful at obtaining a relatively pure Treg population (defined as >50%), while the sixth procedure, due to a technical problem, was not (Treg purity, 42%). Treg cotransfusion significantly delayed death from xenogeneic GVHD in the first five experiments, (p < 0.0001) but not in the sixth experiment. Importantly, none of the mice given enriched Treg alone (Treg group) experienced clinical signs of GVHD, while, interestingly, the CD4+ cells found in these mice 26 days after transplantation were mainly conventional T cells (median CD25+FoxP3+ cells among human CD4+ total cells were only 2.1, 3.1, and 12.2% in spleen, marrow, and blood, respectively).

Conclusions

Infusion of clinical-grade enriched Treg delayed the occurrence of xenogeneic GVHD without inducing toxicity in this murine model.

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