title: Human embryonic stem cell-derived mesenchymal stromal cells ameliorate collagen-induced arthritis by inducing host-derived indoleamine 2,3 dioxygenase.
abstract: BACKGROUND
  The immunosuppressive and anti-inflammatory properties of mesenchymal stromal cells (MSC) have prompted their therapeutic application in several autoimmune diseases, including rheumatoid arthritis. Adult MSC are finite and their clinical use is restricted by the need for long-term expansion protocols that can lead to genomic instability. Inhibition of Smad2/3 signaling in human pluripotent stem cells (hPSC) provides an infinite source of MSC that match the phenotype and functional properties of adult MSC. Here, we test the therapeutic potential of hPSC-MSC of embryonic origin (embryonic stem cell-derived mesenchymal stromal cells, hESC-MSC) in the experimental model of collagen-induced arthritis (CIA).
  
  
  METHODS
  CIA was induced in DBA/1 mice by immunization with type II collagen (CII) in Complete Freund's Adjuvant (CFA). Mice were treated with either a single dose (10(6) cells/mouse) of hESC-MSC on the day of immunization (prophylaxis) or with three doses of hESC-MSC every other day starting on the day of arthritis onset (therapy). Arthritis severity was evaluated daily for six weeks and ten days, respectively. Frequency of Treg (FoxP3(+)), Th1 (IFNγ(+)) and Th17 (IL17(+)) CD4(+) T cells in inguinal lymph nodes (ILN) was quantified by flow cytometry. Serum levels of anti-CII antibodies were determined by ELISA. Detection of hESC-MSC and quantification of murine and human indoleamine 2,3 dioxygenase (IDO1) expression was performed by quantitative real-time PCR. Statistical differences were analyzed by ANOVA and the Mann-Whitney U test.
  
  
  RESULTS
  Administration of hESC-MSC to mice with established arthritis reduced disease severity compared to control-treated mice. Analysis of CD4 T cell populations in treated mice showed an increase in FoxP3(+) Treg and IFNγ(+) Th1 cells but not in Th17 cells in the ILN. Anti-CII antibody levels were not affected by treatment. Migration of hESC-MSC to the ILN in treated mice was associated with the induction of murine IDO1.
  
  
  CONCLUSIONS
  Treatment with hESC-MSC ameliorates CIA by inducing IFNγ(+) Th1 cells and IDO1 in the host. Thus, hESC-MSC can provide an infinite cellular source for treatment of rheumatoid arthritis.
authors.affiliation: Inflammatory and Autoimmune Diseases Group, Hospital 12 de Octubre Research Institute, Madrid, Spain.
authors.forename: Elena
authors.initials: E
authors.lastname: Gonzalo-Gil
authors.affiliation: Inflammatory and Autoimmune Diseases Group, Hospital 12 de Octubre Research Institute, Madrid, Spain.
authors.forename: María J
authors.initials: MJ
authors.lastname: Pérez-Lorenzo
authors.affiliation: Inflammatory and Autoimmune Diseases Group, Hospital 12 de Octubre Research Institute, Madrid, Spain.|Rheumatology Department, Hospital 12 de Octubre, Madrid, Spain.
authors.forename: María
authors.initials: M
authors.lastname: Galindo
authors.affiliation: Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Barcelona, Spain.
authors.forename: Rafael
authors.initials: R
authors.lastname: Díaz de la Guardia
authors.affiliation: Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Barcelona, Spain.
authors.forename: Belén
authors.initials: B
authors.lastname: López-Millán
authors.affiliation: Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Barcelona, Spain.
authors.forename: Clara
authors.initials: C
authors.lastname: Bueno
authors.affiliation: Josep Carreras Leukemia Research Institute, School of Medicine, University of Barcelona, Barcelona, Spain.|Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Spain.
authors.forename: Pablo
authors.initials: P
authors.lastname: Menéndez
authors.affiliation: Inflammatory and Autoimmune Diseases Group, Hospital 12 de Octubre Research Institute, Madrid, Spain.|Rheumatology Department, Hospital 12 de Octubre, Madrid, Spain.
authors.forename: José L
authors.initials: JL
authors.lastname: Pablos
authors.affiliation: Inflammatory and Autoimmune Diseases Group, Hospital 12 de Octubre Research Institute, Madrid, Spain. gcriado@h12o.es.|Inflammatory and Autoimmune Diseases Group, Hospital 12 de Octubre Research Center, Avenida de Córdoba s/n. 28041, Madrid, Spain. gcriado@h12o.es.
authors.forename: Gabriel
authors.identifier: http://orcid.org/0000-0003-4824-0890
authors.initials: G
authors.lastname: Criado
mesh_terms: D000818:Animals; D001169:Arthritis, Experimental / Q000201:enzymology / Q000276:immunology / Q000473:pathology / Q000628:therapy; D001172:Arthritis, Rheumatoid / Q000276:immunology* / Q000473:pathology; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology*; D005434:Flow Cytometry; D064593:Heterografts; D000066449:Human Embryonic Stem Cells / Q000276:immunology / Q000637:transplantation*; D006801:Humans; D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase / Q000378:metabolism*; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods*; D059630:Mesenchymal Stem Cells / Q000276:immunology; D051379:Mice; D008811:Mice, Inbred DBA; D060888:Real-Time Polymerase Chain Reaction
chemical_list: C583637:IDO1 protein, mouse; D050503:Indoleamine-Pyrrole 2,3,-Dioxygenase
keywords: Arthritis; Cytokines; Human embryonic stem cell-derived mesenchymal stromal cells; Indoleamine 2,3 dioxygenase; T cells