title: Developing in vitro expanded CD45RA+ regulatory T cells as an adoptive cell therapy for Crohn's disease.
abstract: OBJECTIVE
  Thymus-derived regulatory T cells (Tregs) mediate dominant peripheral tolerance and treat experimental colitis. Tregs can be expanded from patient blood and were safely used in recent phase 1 studies in graft versus host disease and type 1 diabetes. Treg cell therapy is also conceptually attractive for Crohn's disease (CD). However, barriers exist to this approach. The stability of Tregs expanded from Crohn's blood is unknown. The potential for adoptively transferred Tregs to express interleukin-17 and exacerbate Crohn's lesions is of concern. Mucosal T cells are resistant to Treg-mediated suppression in active CD. The capacity for expanded Tregs to home to gut and lymphoid tissue is unknown.
  
  
  METHODS
  To define the optimum population for Treg cell therapy in CD, CD4(+)CD25(+)CD127(lo)CD45RA(+) and CD4(+)CD25(+)CD127(lo)CD45RA(-) Treg subsets were isolated from patients' blood and expanded in vitro using a workflow that can be readily transferred to a good manufacturing practice background.
  
  
  RESULTS
  Tregs can be expanded from the blood of patients with CD to potential target dose within 22-24 days. Expanded CD45RA(+) Tregs have an epigenetically stable FOXP3 locus and do not convert to a Th17 phenotype in vitro, in contrast to CD45RA(-) Tregs. CD45RA(+) Tregs highly express α4β7 integrin, CD62L and CC motif receptor 7 (CCR7). CD45RA(+) Tregs also home to human small bowel in a C.B-17 severe combined immune deficiency (SCID) xenotransplant model. Importantly, in vitro expansion enhances the suppressive ability of CD45RA(+) Tregs. These cells also suppress activation of lamina propria and mesenteric lymph node lymphocytes isolated from inflamed Crohn's mucosa.
  
  
  CONCLUSIONS
  CD4(+)CD25(+)CD127(lo)CD45RA(+) Tregs may be the most appropriate population from which to expand Tregs for autologous Treg therapy for CD, paving the way for future clinical trials.
authors.affiliation: Medical Research Council Centre for Transplantation, King's College London, London, UK Department of Experimental Immunobiology, King's College London, London, UK National Institute for Health Research Biomedical Research Centre at Guy's and St. Thomas' NHS Foundation Trust and King's College London, London, UK Department of Gastroenterology, Guy's & St Thomas' NHS Foundation Trust, London, UK.
authors.forename: James B
authors.initials: JB
authors.lastname: Canavan
authors.affiliation: Medical Research Council Centre for Transplantation, King's College London, London, UK National Institute for Health Research Biomedical Research Centre at Guy's and St. Thomas' NHS Foundation Trust and King's College London, London, UK Department of Immunoregulation and Immune Intervention, King's College London, London, UK.
authors.forename: Cristiano
authors.initials: C
authors.lastname: Scottà
authors.affiliation: Blizard Institute, Barts and the London School of Medicine and Dentistry, London, UK.
authors.forename: Anna
authors.initials: A
authors.lastname: Vossenkämper
authors.affiliation: Medical Research Council Centre for Transplantation, King's College London, London, UK Department of Experimental Immunobiology, King's College London, London, UK National Institute for Health Research Biomedical Research Centre at Guy's and St. Thomas' NHS Foundation Trust and King's College London, London, UK Department of Gastroenterology, Guy's & St Thomas' NHS Foundation Trust, London, UK.
authors.forename: Rimma
authors.initials: R
authors.lastname: Goldberg
authors.affiliation: Medical Research Council Centre for Transplantation, King's College London, London, UK Department of Experimental Immunobiology, King's College London, London, UK National Institute for Health Research Biomedical Research Centre at Guy's and St. Thomas' NHS Foundation Trust and King's College London, London, UK.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Elder
authors.affiliation: The Koret School of Veterinary Medicine, Hebrew University of Jerusalem, Rehovot, Israel.
authors.forename: Irit
authors.initials: I
authors.lastname: Shoval
authors.affiliation: Medical Research Council Centre for Transplantation, King's College London, London, UK Department of Experimental Immunobiology, King's College London, London, UK National Institute for Health Research Biomedical Research Centre at Guy's and St. Thomas' NHS Foundation Trust and King's College London, London, UK.
authors.forename: Ellen
authors.initials: E
authors.lastname: Marks
authors.affiliation: National Institute for Health Research Biomedical Research Centre at Guy's and St. Thomas' NHS Foundation Trust and King's College London, London, UK Division of Diabetes and Nutritional Sciences, King's College London, London, UK.
authors.forename: Emilie
authors.initials: E
authors.lastname: Stolarczyk
authors.affiliation: Medical Research Council Centre for Transplantation, King's College London, London, UK Department of Experimental Immunobiology, King's College London, London, UK National Institute for Health Research Biomedical Research Centre at Guy's and St. Thomas' NHS Foundation Trust and King's College London, London, UK.
authors.forename: Jonathan W
authors.initials: JW
authors.lastname: Lo
authors.affiliation: Medical Research Council Centre for Transplantation, King's College London, London, UK Department of Experimental Immunobiology, King's College London, London, UK National Institute for Health Research Biomedical Research Centre at Guy's and St. Thomas' NHS Foundation Trust and King's College London, London, UK Department of Gastroenterology, Guy's & St Thomas' NHS Foundation Trust, London, UK.
authors.forename: Nick
authors.initials: N
authors.lastname: Powell
authors.affiliation: Medical Research Council Centre for Transplantation, King's College London, London, UK National Institute for Health Research Biomedical Research Centre at Guy's and St. Thomas' NHS Foundation Trust and King's College London, London, UK Department of Immunoregulation and Immune Intervention, King's College London, London, UK.
authors.forename: Henrieta
authors.initials: H
authors.lastname: Fazekasova
authors.affiliation: Department of Gastroenterology, Guy's & St Thomas' NHS Foundation Trust, London, UK.
authors.forename: Peter M
authors.initials: PM
authors.lastname: Irving
authors.affiliation: Department of Gastroenterology, Guy's & St Thomas' NHS Foundation Trust, London, UK.
authors.forename: Jeremy D
authors.initials: JD
authors.lastname: Sanderson
authors.affiliation: National Institute for Health Research Biomedical Research Centre at Guy's and St. Thomas' NHS Foundation Trust and King's College London, London, UK Division of Diabetes and Nutritional Sciences, King's College London, London, UK.
authors.forename: Jane K
authors.initials: JK
authors.lastname: Howard
authors.affiliation: Department of Obstetrics & Gynaecology, Hadassah University Hospital, Jerusalem, Israel.
authors.forename: Simcha
authors.initials: S
authors.lastname: Yagel
authors.affiliation: Medical Research Council Centre for Transplantation, King's College London, London, UK National Institute for Health Research Biomedical Research Centre at Guy's and St. Thomas' NHS Foundation Trust and King's College London, London, UK Department of Immunoregulation and Immune Intervention, King's College London, London, UK.
authors.forename: Behdad
authors.initials: B
authors.lastname: Afzali
authors.affiliation: Blizard Institute, Barts and the London School of Medicine and Dentistry, London, UK.
authors.forename: Thomas T
authors.initials: TT
authors.lastname: MacDonald
authors.affiliation: Medical Research Council Centre for Transplantation, King's College London, London, UK Department of Experimental Immunobiology, King's College London, London, UK National Institute for Health Research Biomedical Research Centre at Guy's and St. Thomas' NHS Foundation Trust and King's College London, London, UK.
authors.forename: Maria P
authors.initials: MP
authors.lastname: Hernandez-Fuentes
authors.affiliation: The Koret School of Veterinary Medicine, Hebrew University of Jerusalem, Rehovot, Israel.
authors.forename: Nahum Y
authors.initials: NY
authors.lastname: Shpigel
authors.affiliation: Medical Research Council Centre for Transplantation, King's College London, London, UK National Institute for Health Research Biomedical Research Centre at Guy's and St. Thomas' NHS Foundation Trust and King's College London, London, UK Department of Immunoregulation and Immune Intervention, King's College London, London, UK.
authors.forename: Giovanna
authors.initials: G
authors.lastname: Lombardi
authors.affiliation: Medical Research Council Centre for Transplantation, King's College London, London, UK Department of Experimental Immunobiology, King's College London, London, UK National Institute for Health Research Biomedical Research Centre at Guy's and St. Thomas' NHS Foundation Trust and King's College London, London, UK.
authors.forename: Graham M
authors.initials: GM
authors.lastname: Lord
mesh_terms: D019264:Adoptive Transfer; D000818:Animals; D064987:Cell- and Tissue-Based Therapy / Q000379:methods*; D003424:Crohn Disease / Q000276:immunology / Q000628:therapy*; D019175:DNA Methylation; D004797:Enzyme-Linked Immunosorbent Assay; D051858:Forkhead Transcription Factors / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D020381:Interleukin-17 / Q000378:metabolism; D017493:Leukocyte Common Antigens / Q000276:immunology; D051379:Mice; D016513:Mice, SCID; D010641:Phenotype; D016133:Polymerase Chain Reaction; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D014183:Transplantation, Heterologous
chemical_list: C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D020381:Interleukin-17; D017493:Leukocyte Common Antigens
keywords: IBD; IBD BASIC RESEARCH; IBD CLINICAL; IMMUNOTHERAPY; INTESTINAL T CELLS