title: A microRNA profile of human CD8(+) regulatory T cells and characterization of the effects of microRNAs on Treg cell-associated genes.
abstract: BACKGROUND
  Recently, regulatory T (Treg) cells have gained interest in the fields of immunopathology, transplantation and oncoimmunology. Here, we investigated the microRNA expression profile of human natural CD8(+)CD25(+) Treg cells and the impact of microRNAs on molecules associated with immune regulation.
  
  
  METHODS
  We purified human natural CD8(+) Treg cells and assessed the expression of FOXP3 and CTLA-4 by flow cytometry. We have also tested the ex vivo suppressive capacity of these cells in mixed leukocyte reactions. Using TaqMan low-density arrays and microRNA qPCR for validation, we could identify a microRNA 'signature' for CD8(+)CD25(+)FOXP3(+)CTLA-4(+) natural Treg cells. We used the 'TargetScan' and 'miRBase' bioinformatics programs to identify potential target sites for these microRNAs in the 3'-UTR of important Treg cell-associated genes.
  
  
  RESULTS
  The human CD8(+)CD25(+) natural Treg cell microRNA signature includes 10 differentially expressed microRNAs. We demonstrated an impact of this signature on Treg cell biology by showing specific regulation of FOXP3, CTLA-4 and GARP gene expression by microRNA using site-directed mutagenesis and a dual-luciferase reporter assay. Furthermore, we used microRNA transduction experiments to demonstrate that these microRNAs impacted their target genes in human primary Treg cells ex vivo.
  
  
  CONCLUSIONS
  We are examining the biological relevance of this 'signature' by studying its impact on other important Treg cell-associated genes. These efforts could result in a better understanding of the regulation of Treg cell function and might reveal new targets for immunotherapy in immune disorders and cancer.
authors.affiliation: Experimental Hematology, Institut Jules Bordet, Université Libre de Bruxelles, 121, Boulevard de Waterloo, 1000, Bruxelles, Belgium. Fadi.Jebbawi@ulb.ac.be.
authors.forename: Fadi
authors.initials: F
authors.lastname: Jebbawi
authors.affiliation: Experimental Hematology, Institut Jules Bordet, Université Libre de Bruxelles, 121, Boulevard de Waterloo, 1000, Bruxelles, Belgium. hfayyadk@ulb.ac.be.
authors.forename: Hussein
authors.initials: H
authors.lastname: Fayyad-Kazan
authors.affiliation: Experimental Hematology, Institut Jules Bordet, Université Libre de Bruxelles, 121, Boulevard de Waterloo, 1000, Bruxelles, Belgium. mmerimi@ulb.ac.be.
authors.forename: Makram
authors.initials: M
authors.lastname: Merimi
authors.affiliation: Experimental Hematology, Institut Jules Bordet, Université Libre de Bruxelles, 121, Boulevard de Waterloo, 1000, Bruxelles, Belgium. philippe.lewalle@bordet.be.
authors.forename: Philippe
authors.initials: P
authors.lastname: Lewalle
authors.affiliation: Department of Gynecology and Obstetrics, Clinique Saint-Pierre, Ottignies, Belgium. je.verougstraete@clinique-saint-pierre.be.
authors.forename: Jean-Christophe
authors.initials: JC
authors.lastname: Verougstraete
authors.affiliation: Experimental Hematology, Institut Jules Bordet, Université Libre de Bruxelles, 121, Boulevard de Waterloo, 1000, Bruxelles, Belgium. oleo@ulb.ac.be.
authors.forename: Oberdan
authors.initials: O
authors.lastname: Leo
authors.affiliation: Ludwig Center for Cancer Research of the University of Lausanne, Lausanne, Switzerland. pedro.romero@unil.ch.
authors.forename: Pedro
authors.initials: P
authors.lastname: Romero
authors.affiliation: Experimental Hematology, Institut Jules Bordet, Université Libre de Bruxelles, 121, Boulevard de Waterloo, 1000, Bruxelles, Belgium. arsene.burny@guest.ulg.ac.be.
authors.forename: Arsene
authors.initials: A
authors.lastname: Burny
authors.affiliation: Department of Biochemistry, Laboratory of Immunology, EDST-PRASE, Faculty of Sciences, Lebanese University, Hadath-Beirut, Lebanon. bbadran@ulb.ac.be.
authors.forename: Bassam
authors.initials: B
authors.lastname: Badran
authors.affiliation: Experimental Hematology, Institut Jules Bordet, Université Libre de Bruxelles, 121, Boulevard de Waterloo, 1000, Bruxelles, Belgium. pmartiat@ulb.ac.be.
authors.forename: Philippe
authors.initials: P
authors.lastname: Martiat
authors.affiliation: Experimental Hematology, Institut Jules Bordet, Université Libre de Bruxelles, 121, Boulevard de Waterloo, 1000, Bruxelles, Belgium. mrouas@ulb.ac.be.
authors.forename: Redouane
authors.initials: R
authors.lastname: Rouas
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D015703:Antigens, CD / Q000378:metabolism; D018414:CD8-Positive T-Lymphocytes / Q000378:metabolism*; D060908:CTLA-4 Antigen / Q000378:metabolism; D049109:Cell Proliferation; D002469:Cell Separation; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006367:HeLa Cells; D006801:Humans; D016086:Lentivirus / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D050378:T-Lymphocytes, Regulatory / Q000378:metabolism*; D014158:Transcription, Genetic; D014161:Transduction, Genetic
chemical_list: D020413:3' Untranslated Regions; D015703:Antigens, CD; D060908:CTLA-4 Antigen; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; C087215:LRRC32 protein, human; D008565:Membrane Proteins; D035683:MicroRNAs
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