title: [miR-126 knockdown enhances the activity of murine CD4+ T cells in vivo and promotes their differentiation into Th1 cells].
abstract: OBJECTIVE
  To investigate the change of CD4(+) T cell activity in microRNA-126 (miR-126) knockdown (KD) mice and explore its significance.
  
  
  METHODS
  The expression level of mature miR-126 in CD4(+) CD62L(+) T cells purified by magnetic-activated cell sorting (MACS) was analyzed by real-time PCR using specific probe. Furthermore, the expression levels of CD69, CD62L and CD44 molecules, as well as intracellular proliferating nuclear antigen Ki-67, in CD4(+) T cells in miR-126 KD mice were detected by fluorescence-activated cell sorting (FACS). Moreover, the apoptosis of CD4(+) T cells was analyzed by annexin V/PI staining assay combined with flow cytometry. Finally, the relative expressions of function-related cytokines including interleukine 4 (IL-4), IL-10, IL-12, transforming growth factor (TGF-β), interferon (IFN-γ) and tumor necrosis factor (TNF-α) in CD4(+) T cells were determined by real-time PCR.
  
  
  RESULTS
  Compared with wild-type (WT) mice, the expression level of mature miR-126 in CD4(+) T cells in miR-126 KD mice was dramatically reduced. Furthermore, the proportion of CD62L(+) in CD4(+) T cells also decreased significantly, while the proportions of CD69(+), CD44(+) and Ki-67(+) cells were remarkably elevated. Meanwhile, the apoptosis proportion of CD4(+) T cells in vivo dropped dramatically in miR-126 KD mice. Finally, the mRNA expressions of IL-4 and IL-10 in CD4(+)T cells were significantly downregulated, but IL-12, TGF-β, TNF-α and IFN-γ mRNAs were obviously up-regulated.
  
  
  CONCLUSIONS
  miR-126 knockdown could significantly enhance the functional activity of CD4(+) T cells in vivo and promote cell differentiation into Th1 cells.
authors.affiliation: Department of Immunology, Zunyi Medical College, Zunyi 563000, China.
authors.forename: Panpan
authors.initials: P
authors.lastname: Cui
authors.affiliation: Department of Immunology, Zunyi Medical College, Zunyi 563000, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Hu
authors.affiliation: Department of Immunology, Zunyi Medical College, Zunyi 563000, China.
authors.forename: Yijing
authors.initials: Y
authors.lastname: Tao
authors.affiliation: Department of Immunology, Zunyi Medical College, Zunyi 563000, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Chen
authors.affiliation: Department of Immunology, Zunyi Medical College, Zunyi 563000, China.
authors.forename: Juanjuan
authors.initials: J
authors.lastname: Zhao
authors.affiliation: Department of Immunology, Zunyi Medical College, Zunyi 563000, China.
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Guo
authors.affiliation: Department of Medical Physics, Zunyi Medical College, Zunyi 563000, China.
authors.forename: Ya
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: Department of Immunology, Zunyi Medical College, Zunyi 563000, China. *Corresponding author, E-mail: xulinzhouya@163.com.
authors.forename: Lin
authors.initials: L
authors.lastname: Xu
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D000945:Antigens, Differentiation, T-Lymphocyte / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D002454:Cell Differentiation / Q000235:genetics* / Q000276:immunology; D016207:Cytokines / Q000235:genetics; D005434:Flow Cytometry; D015870:Gene Expression; D055785:Gene Knockdown Techniques / Q000379:methods*; D018960:Hyaluronan Receptors / Q000378:metabolism; D018189:Immunomagnetic Separation; D019394:Ki-67 Antigen / Q000378:metabolism; D019041:L-Selectin / Q000378:metabolism; D037181:Lectins, C-Type / Q000378:metabolism; D051379:Mice; D035683:MicroRNAs / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018417:Th1 Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D000099192:CD69 Antigens
chemical_list: D015703:Antigens, CD; D000945:Antigens, Differentiation, T-Lymphocyte; D016207:Cytokines; D018960:Hyaluronan Receptors; D019394:Ki-67 Antigen; D019041:L-Selectin; D037181:Lectins, C-Type; D035683:MicroRNAs; D000099192:CD69 Antigens; C545201:MIRN126 microRNA, mouse