title: [Small hairpin RNA silencing Tim-3 affects peripheral blood Th1 and Th17 cells differentiation in asthmatic mice].
abstract: OBJECTIVE
  To investigate the effects of down-regulating Tim-3 gene in the peripheral blood mononuclear cells (PBMCs) of an asthmatic mouse model by short hairpin RNA (shRNA) and to explore the effect of Tim-3 on Th1 and Th17 cell differentiation.
  
  
  METHODS
  An asthmatic murine model was established by ovalbumin sensitization and challenge. PBMCs were isolated from asthmatic mice and transfected by shRNA targeting Tim-3 gene. The mRNA and protein expressions of Tim-3 were detected by quantitative PCR and Western blot. Flow cytometry analysis was performed to determine the levels of Th1 and Th17, and ELISA was performed to determine concentrations of IFN-γ, IL-4 and IL-17 in the supernatant.
  
  
  RESULTS
  Tim-3 mRNA expression in PBMCs was significantly increased in asthmatic mice. The mRNA and protein expression of Tim-3 decreased significantly in the shRNA group. Compared with the negative groups, Th1 cell levels increased and Th17 cell levels decreased significantly in the asthmatic groups after Tim-3 shRNA interference. In the Tim-3 shRNA interference groups concentrations of IFN-γ increased significantly while IL-17 decreased significantly.
  
  
  CONCLUSIONS
  Specific Tim-3 shRNA effectively silences the expression of Tim-3 and change in Tim-3 expression could affect T cell differentiation.
authors.affiliation: Department of Pulmonary Medicine, Wuhan Childern's Hospital, Wuhan, China. lusi74@163.com
authors.forename: Xiao-Xia
authors.initials: XX
authors.lastname: Lu
authors.forename: Jia-Li
authors.initials: JL
authors.lastname: Xu
authors.forename: Zong-Qi
authors.initials: ZQ
authors.lastname: Dong
authors.forename: Peng
authors.initials: P
authors.lastname: Chen
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
mesh_terms: D000818:Animals; D001249:Asthma / Q000276:immunology / Q000628:therapy*; D002454:Cell Differentiation; D005260:Female; D020868:Gene Silencing; D000072597:Hepatitis A Virus Cellular Receptor 2; D051379:Mice; D008807:Mice, Inbred BALB C; D034741:RNA, Small Interfering / Q000235:genetics*; D011991:Receptors, Virus / Q000037:antagonists & inhibitors / Q000235:genetics*; D018417:Th1 Cells / Q000166:cytology*; D058504:Th17 Cells / Q000166:cytology*
chemical_list: C442241:Havcr2 protein, mouse; D000072597:Hepatitis A Virus Cellular Receptor 2; D034741:RNA, Small Interfering; D011991:Receptors, Virus