title: Obtaining regulatory T cells from uraemic patients awaiting kidney transplantation for use in clinical trials.
abstract: Adoptive transfer of regulatory T cells (T(regs)) has been proposed for use as a cellular therapy to induce transplantation tolerance. Preclinical data are encouraging, and clinical trials with T(reg) therapy are anticipated. In this study, we investigate different strategies for the isolation and expansion of CD4(+) CD25(high) CD127(low) T(regs) from uraemic patients. We use allogeneic dendritic cells (DCs) as feeder cells for the expansion and compare T(reg) preparations isolated by either fluorescence activated cell sorting (FACS) or magnetic activated cell sorting (MACS) that have been expanded subsequently with either mature or tolerogenic DCs. Expanded T(reg) preparations have been characterized by their purity, cytokine production and in-vitro suppressive ability. The results show that T(reg) preparations can be isolated from uraemic patients by both FACS and MACS. Also, the type of feeder cells used in the expansion affects both the purity and the functional properties of the T(reg) preparations. In particular, FACS-sorted T(reg) preparations expanded with mature DCs secrete more interleukin (IL)-10 and granzyme B than FACS-sorted T(reg) preparations expanded with tolerogenic DCs. This is a direct comparison between different isolation techniques and expansion protocols with T(regs) from uraemic patients that may guide future efforts to produce clinical-grade T(regs) for use in kidney transplantation.
authors.affiliation: Department of Surgical Sciences, Section of Transplantation Surgery, Uppsala University, Uppsala, Sweden. david.berglund@surgsci.uu.se
authors.forename: D
authors.initials: D
authors.lastname: Berglund
authors.forename: M
authors.initials: M
authors.lastname: Karlsson
authors.forename: A-R
authors.initials: AR
authors.lastname: Biglarnia
authors.forename: T
authors.initials: T
authors.lastname: Lorant
authors.forename: G
authors.initials: G
authors.lastname: Tufveson
authors.forename: O
authors.initials: O
authors.lastname: Korsgren
authors.forename: B
authors.initials: B
authors.lastname: Carlsson
mesh_terms: D015704:CD4 Antigens / Q000378:metabolism; D018929:Cell Culture Techniques / Q000379:methods*; D048708:Cell Growth Processes; D002469:Cell Separation; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D003713:Dendritic Cells / Q000276:immunology*; D005434:Flow Cytometry; D006680:HLA Antigens / Q000276:immunology; D006801:Humans; D007165:Immunosuppression Therapy; D016219:Immunotherapy, Adoptive / Q000379:methods*; D053645:Interleukin-2 Receptor alpha Subunit / Q000378:metabolism; D053699:Interleukin-7 Receptor alpha Subunit / Q000378:metabolism; D007674:Kidney Diseases / Q000276:immunology* / Q000628:therapy; D016030:Kidney Transplantation; D017410:Practice Guidelines as Topic; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D023001:Transplantation Tolerance; D014850:Waiting Lists
chemical_list: D015704:CD4 Antigens; D016207:Cytokines; D006680:HLA Antigens; D053645:Interleukin-2 Receptor alpha Subunit; D053699:Interleukin-7 Receptor alpha Subunit
keywords: cell therapy; clinical trials; kidney transplantation; regulatory T cells; tolerance