title: Ex vivo expansion of antigen-specific CD4+CD25+ regulatory T cells from autologous naïve CD4+ T cells of multiple sclerosis patients as a potential therapeutic approach.
abstract: CD4+CD25+ regulatory T cells (Tregs) have been found to have a decreased effector function in patients with multiple sclerosis (MS). In this study, we co-cultured naïve CD4+ T cells of MS patients with myelin basic protein (MBP)85-99 peptide as specific antigen and allogenic B cells as antigen-presenting cells, in an attempt to generate adequate antigen-specific CD4+CD25+ Tregs with normal or improved immune function.
  
  
  
  Naïve CD4+ T cells were isolated from peripheral blood mononuclear cells (PBMCs) from patients with MS (n=5) and healthy controls (HC, n=5). Furthermore, these purified naive CD4+ T cells were co-cultured with the CD40-activated B cells and MBP85-99 peptide to induce MBP-reactive CD4+CD25highCD127low Tregs. After harvesting these Tregs via a flow sorter, real-time PCR and mixed lymphocyte reaction (MLR) assay were performed to characterize cellular immune function. Supernatant interleukin (IL)-10 and transforming growth factor (TGF)-β1 protein levels were detected by an enzyme-linked immunosorbent assay (ELISA).
  
  
  
  With this method, the frequency of CD4+CD25highCD127low Tregs in CD4+ T cells was 3.5%-6%. In both MS and HC groups, there were relatively lower proliferation indices (PI) of MLR assay but higher supernatant IL-10 and TGF-β1 levels in the presence of MBP than those in the presence of other control antigens, where no significant differences were found.
  
  
  
  Via the ex vivo culture, adequate MBP-reactive CD4+CD25+ Tregsderived from autologous naïve CD4+ T cells of MS patients, were obtained and returned to normal without immune defects, and even upregulated their immunosuppressive function mostly through the elevated release of IL-10 and TGF-β1.
authors.affiliation: Department of Neurology, Peking University People's Hospital, Beijing, China. guangzhi2002@hotmail.com.
authors.forename: Y-J
authors.initials: YJ
authors.lastname: Xiang
authors.forename: M
authors.initials: M
authors.lastname: Ren
authors.forename: H
authors.initials: H
authors.lastname: Jiang
authors.forename: T-T
authors.initials: TT
authors.lastname: Yang
authors.forename: Y
authors.initials: Y
authors.lastname: He
authors.forename: D-H
authors.initials: DH
authors.lastname: Ao
authors.forename: Y-Y
authors.initials: YY
authors.lastname: Wang
authors.forename: Q
authors.initials: Q
authors.lastname: Zhang
authors.forename: X-J
authors.initials: XJ
authors.lastname: He
authors.forename: X-G
authors.initials: XG
authors.lastname: Gao
authors.forename: G-Z
authors.initials: GZ
authors.lastname: Liu
mesh_terms: D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism; D051858:Forkhead Transcription Factors / Q000378:metabolism; D006801:Humans; D053645:Interleukin-2 Receptor alpha Subunit / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D009103:Multiple Sclerosis / Q000276:immunology* / Q000628:therapy*; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*
chemical_list: D051858:Forkhead Transcription Factors; C508017:IL2RA protein, human; D053645:Interleukin-2 Receptor alpha Subunit