title: Dysregulation of anergy-related factors involved in regulatory T cells defects in Systemic Lupus Erythematosus patients: Rapamycin and Vitamin D efficacy in restoring regulatory T cells.
abstract: OBJECTIVE
  Systemic Lupus Erythematosus (SLE) patients display dysfunctions in T cell activation and anergy. Therefore the aims of our study were to explore the expression of anergy-related factors in CD4+T cells in relationship with regulatory T cells (Tregs) frequency in SLE patients and to identify strategies to redress these defects.
  
  
  METHODS
  Casitas B-cell lymphoma b (Cbl-b) and 'gene related to anergy in lymphocytes' (GRAIL) proteins were analyzed in peripheral blood mononuclear cells (PBMCs) from SLE patients and healthy donors (HD) by immunoblotting. cbl-b, grail, growth response factors (egr)2 and egr3 messenger RNAs (mRNAs) were evaluated by real-time polymerase chain reaction in SLE and HD PBMCs and CD4+T cells. Phenotypic and functional characterization of CD4+T cells was performed by flow cytometry. Tregs expansion protocol consisted in culturing CD4+T cells for 14 or 21 days of experimental activation with anti-CD3 and anti-CD28 monoclonal antibodies, human recombinant interleukin (hrIL)-2, in the absence or presence of rapamycin (Rapa) or 1,25-(OH)2D3 (vitamin D: VitD).
  
  
  RESULTS
  SLE PBMCs expressed low levels of Cbl-b and GRAIL proteins. Both SLE PBMCs and CD4+T cells expressed low levels of egr2/3 mRNAs. SLE patients had a reduced number of Tregs with impaired suppressive activity. An association between egr2 mRNA level in CD4+T cells and Tregs percentage was identified. Experimental activation of CD4+T cells in the presence of hrIL-2 and Rapa or VitD induced the expansion of SLE Tregs. However, on long-term, only Rapa exposure of SLE CD4+T cells yielded high numbers of Tregs with sustained suppressive activity.
  
  
  CONCLUSIONS
  Our results suggest a new strategy to correct defects in CD4+T cell tolerance mechanisms that may prove beneficial in SLE.
authors.affiliation: Cantacuzino National Institute of Research and Development for Microbiology and Immunology, Cellular and Molecular Immunity Laboratory, Bucharest, Romania.|Department of Biochemistry and Molecular Biology, Faculty of Biology, University of Bucharest, Bucharest, Romania.
authors.forename: Leontina M
authors.initials: LM
authors.lastname: Banica
authors.affiliation: Cantacuzino National Institute of Research and Development for Microbiology and Immunology, Cellular and Molecular Immunity Laboratory, Bucharest, Romania.
authors.forename: Alina N
authors.initials: AN
authors.lastname: Besliu
authors.affiliation: Cantacuzino National Institute of Research and Development for Microbiology and Immunology, Cellular and Molecular Immunity Laboratory, Bucharest, Romania.
authors.forename: Gina C
authors.initials: GC
authors.lastname: Pistol
authors.affiliation: Cantacuzino National Institute of Research and Development for Microbiology and Immunology, Cellular and Molecular Immunity Laboratory, Bucharest, Romania.
authors.forename: Crina
authors.initials: C
authors.lastname: Stavaru
authors.affiliation: Department of Rheumatology and Internal Medicine, Sf. Maria Clinical Hospital, Bucharest, Romania.
authors.forename: Violeta
authors.initials: V
authors.lastname: Vlad
authors.affiliation: Department of Rheumatology and Internal Medicine, Sf. Maria Clinical Hospital, Bucharest, Romania.|Carol Davila University of Medicine and Pharmacy, Bucharest, Romania.
authors.forename: Denisa
authors.initials: D
authors.lastname: Predeteanu
authors.affiliation: Department of Rheumatology and Internal Medicine, Sf. Maria Clinical Hospital, Bucharest, Romania.|Carol Davila University of Medicine and Pharmacy, Bucharest, Romania.
authors.forename: Ruxandra
authors.initials: R
authors.lastname: Ionescu
authors.affiliation: Cantacuzino National Institute of Research and Development for Microbiology and Immunology, Cellular and Molecular Immunity Laboratory, Bucharest, Romania.
authors.forename: Maria
authors.initials: M
authors.lastname: Stefanescu
authors.affiliation: Cantacuzino National Institute of Research and Development for Microbiology and Immunology, Cellular and Molecular Immunity Laboratory, Bucharest, Romania.
authors.forename: Cristiana
authors.initials: C
authors.lastname: Matache
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000097:blood / Q000235:genetics; D002117:Calcitriol / Q000494:pharmacology*; D016022:Case-Control Studies; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D017635:Clonal Anergy / Q000187:drug effects*; D051767:Early Growth Response Protein 2 / Q000097:blood / Q000235:genetics; D051776:Early Growth Response Protein 3 / Q000097:blood / Q000235:genetics; D006801:Humans; D007155:Immunologic Factors / Q000494:pharmacology*; D008180:Lupus Erythematosus, Systemic / Q000097:blood / Q000188:drug therapy* / Q000276:immunology; D008213:Lymphocyte Activation / Q000187:drug effects; D010641:Phenotype; D050721:Proto-Oncogene Proteins c-cbl / Q000097:blood / Q000235:genetics; D012333:RNA, Messenger / Q000097:blood / Q000235:genetics; D017634:Self Tolerance / Q000187:drug effects; D020123:Sirolimus / Q000494:pharmacology*; D050378:T-Lymphocytes, Regulatory / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism; D013997:Time Factors; D044767:Ubiquitin-Protein Ligases / Q000097:blood / Q000235:genetics
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C497326:EGR2 protein, human; C079945:EGR3 protein, human; D051767:Early Growth Response Protein 2; D007155:Immunologic Factors; D012333:RNA, Messenger; D051776:Early Growth Response Protein 3; C492211:CBLB protein, human; D050721:Proto-Oncogene Proteins c-cbl; C520250:RNF128 protein, human; D044767:Ubiquitin-Protein Ligases; D002117:Calcitriol; D020123:Sirolimus
keywords: anergy-related factors; rapamycin; regulatory T cells; systemic lupus erythematosus; vitamin D