title: Generation of Donor-specific T Regulatory Type 1 Cells From Patients on Dialysis for Cell Therapy After Kidney Transplantation.
abstract: BACKGROUND
  T regulatory type 1 (Tr1) cell-mediated induction of tolerance in preclinical models of transplantation is remarkably effective. The clinical application of such a therapy in patients on dialysis undergoing kidney transplantation should take into account the possible alterations of the immune system observed in these patients. Herein, we aimed at testing the ability to generate donor-specific Tr1 cell-enriched lymphocytes from patients on dialysis on the waiting list for kidney transplantation.
  
  
  METHODS
  The Tr1 cell-enriched lymphocytes were generated by coculturing interleukin-10-producing dendritic cells obtained from healthy donors with peripheral blood mononuclear cells (PBMCs) of patients on dialysis, following the same protocol used in a previous cell therapy clinical trial to prevent graft-versus-host disease. Alternatively, purified CD4(+) T cells were used instead of total PBMCs. The ability to generate clinical-grade Tr1 cell-enriched products was defined by testing the reduced response to restimulation with mature dendritic cells generated from the original donor (i.e., anergy assay).
  
  
  RESULTS
  The Tr1 cell-enriched medicinal products generated from PBMCs of patients on dialysis showed a low anergic phenotype, incompatible with their eventual clinical application. This was irrespective of HLA matching with the donor or the intrinsically reduced ability to proliferate in response to alloantigens. On the contrary, the use of purified CD4(+) T cells isolated from patients on dialysis led to the generation of a highly anergic donor-specific medicinal product containing an average of 10% Tr1 cells.
  
  
  CONCLUSIONS
  The Tr1 cell-enriched medicinal products can be efficiently generated from patients on dialysis by carefully tailoring the protocol on the patients' immunological characteristics.
authors.affiliation: 1 Transplant Unit, Ospedale San Raffaele, Milan, Italy. 2 Currently, Laboratory for Translational Immunology, Universiteit Medisch Centrum Utrecht, Utrecht, The Netherlands; formerly, Diabetes Research Institute IRCCS San Raffaele Scientific Institute, Milan, Italy. 3 Diabetes Research Institute IRCCS San Raffaele Scientific Institute, Milan, Italy. 4 Nephrology and Dialysis Unit, Ospedale San Raffaele, Milan, Italy. 5 Universita' Vita-Salute San Raffaele, Milan, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Petrelli
authors.forename: Eleonora
authors.initials: E
authors.lastname: Tresoldi
authors.forename: Bechara G
authors.initials: BG
authors.lastname: Mfarrej
authors.forename: Alessia
authors.initials: A
authors.lastname: Paganelli
authors.forename: Donatella
authors.initials: D
authors.lastname: Spotti
authors.forename: Rossana
authors.initials: R
authors.lastname: Caldara
authors.forename: Antonio
authors.initials: A
authors.lastname: Secchi
authors.forename: Manuela
authors.initials: M
authors.lastname: Battaglia
mesh_terms: D000328:Adult; D016022:Case-Control Studies; D002450:Cell Communication; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D017635:Clonal Anergy; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000276:immunology / Q000378:metabolism; D005260:Female; D006801:Humans; D007108:Immune Tolerance; D016753:Interleukin-10 / Q000276:immunology / Q000378:metabolism; D007676:Kidney Failure, Chronic / Q000175:diagnosis / Q000276:immunology / Q000628:therapy*; D016030:Kidney Transplantation; D008297:Male; D008875:Middle Aged; D010641:Phenotype; D006435:Renal Dialysis; D015398:Signal Transduction; D050378:T-Lymphocytes, Regulatory / Q000276:immunology* / Q000378:metabolism; D014850:Waiting Lists
chemical_list: C508609:IL10 protein, human; D016753:Interleukin-10