Generation of donor-specific Tr1 cells to be used after kidney transplantation and definition of the timing of their in vivo infusion in the presence of immunosuppression

Abstract

BackgroundOperational tolerance is an alternative to lifelong immunosuppression after transplantation. One strategy to achieve tolerance is by T regulatory cells. Safety and feasibility of a T regulatory type 1 (Tr1)-cell—based therapy to prevent graft versus host disease in patients with hematological malignancies has been already proven. We are now planning to perform a Tr1-cell—based therapy after kidney transplantation.MethodsUpon tailoring the lab-grade protocol to patients on dialysis, aims of the current work were to develop a clinical-grade compatible protocol to generate a donor-specific Tr1-cell—enriched medicinal product (named T10 cells) and to test the Tr1-cell sensitivity to standard immunosuppression in vivo to define the best timing of cell infusion.ResultsWe developed a medicinal product that was enriched in Tr1 cells, anergic to donor-cell stimulation, able to suppress proliferation upon donor- but not third-party stimulation in vitro, and stable upon cryopreservation. The protocol was reproducible upon up scaling to leukapheresis from patients on dialysis and was effective in yielding the expected number of T10 cells necessary for the planned infusions. The tolerogenic gene signature of circulating Tr1 cells was minimally compromised in kidney transplant recipients under standard immunosuppression and it eventually started to recover 36 weeks post-transplantation, providing rationale for selecting the timings of the cell infusions.ConclusionsThese data provide solid ground for proceeding with the trial and establish robust rationale for defining the correct timing of cell infusion during concomitant immunosuppressive treatment.Electronic supplementary materialThe online version of this article (doi:10.1186/s12967-017-1133-8) contains supplementary material, which is available to authorized users.

The Biology of T Regulatory Type 1 Cells and Their Therapeutic Application in Immune-Mediated Diseases

“…generated in vitro can be purified from bulk cultures using CD49b and LAG-3 antibodies (Amodio et al, 2015;Gagliani et al, 2013b;Mfarrej et al, 2017;Petrelli et al, 2015). The frequency of CD4 + CD45RA -CD49b + LAG-3 + cells correlates with the expression of IL-10 in human (Amodio et al, 2015;Mfarrej et al, 2017;Petrelli et al, 2015;Sutavani et al, 2013;Tousa et al, 2017;Yao et al, 2015;Tousa et al, 2017), and with induction of immunological tolerance in different preclinical and clinical settings Gagliani et al, 2013b;Kim et al, 2018;Koch et al, 2015;Zhu et al, 2018). Using single-cell mass cytometry, we have shown that CD49b and LAG-3 were sufficient to identify a population of CD4 + CD45RAcells, which is distinct from Th1, Th2, Th17, Tfh, and Treg cells.…”

Regulatory T cells in renal disease

“…In addition, other suppressive cells not belonging to the Th lineage include CD3 + CD4 − CD8 − double‐negative T cells, Qa‐1‐restricted CD8 Tregs and CD28 lo CD8 + Tregs . While these additional regulatory T‐cell subsets have not been as well studied as the conventional CD4 + Foxp3 + Tregs that are discussed in this review, there is evidence for their protective involvement in renal disease …”

“…These protocols have been generating cells that maintain their phenotype including demethylation of the FOXP3 TSDR . Furthermore, Tr1 cells can also be generated from patients with end‐stage kidney disease who are on dialysis …”

“…[127][128][129] While these additional regulatory T-cell subsets have not been as well studied as the conventional CD4 + Foxp3 + Tregs that are discussed in this review, there is evidence for their protective involvement in renal disease. 27,123,[130][131][132]…”

Harnessing the power of regulatory T‐cells to control autoimmune diabetes: overview and perspective

“…94 DC10-expanded Tr1 cells have also been selected to be a part of the ONE Study to assess the safety, clinical practicality and therapeutic efficacy in kidney transplanted recipients. 95,96 Despite the limited use of Tr1 in present clinical studies, with increased recognition of their importance in controlling T1D and other autoimmune and inflammatory diseases, 41,43,97 a broader use of Tr1 cells might be forthcoming.…”