title: A preliminary study towards downregulation of murine bone marrow eosinophilopoiesis mediated by small molecule inhibition of interleukin-5 receptor alpha gene in vitro.
abstract: BACKGROUND
  Bone marrow eosinophilopoiesis induced by IL-5 makes a major contribution to eosinophilic airway inflammation in asthma. Bone marrow CD(34)(+) cells expressing IL-5Ralpha may be eosinophil progenitors. However, research on the effect of blocking IL-5Ralpha expression on bone marrow eosinophilopoiesis has seldom been reported.
  
  
  OBJECTIVE
  To explore the effect of inhibiting IL-5Ralpha expression with IL-5Ralpha short hairpin RNA-expressing vector on murine bone marrow eosinophilopoiesisin vitro.
  
  
  METHODS
  We constructed 4 kinds of plasmid vectors that could express small molecule inhibition, short hairpin RNA, which targeted IL-5Ralpha (P-IL-5Ralpha), and selected an effective one by transfecting B lymphoma cells in vitro. We also constructed an adenovirus vector which was inserted into an effective template sequence (Ad-IL-5Ralpha). The bone marrow cells were obtained from healthy Balb/c mice, and cultured and transfected by Ad-IL-5Ralpha in vitro. The expression of IL-5Ralpha and the count of newly produced eosinophils were detected in the cultured bone marrow cells.
  
  
  RESULTS
  We found that P-IL-5Ralpha-3 targeted at the sequence of CAG CTG CCT GGT TCG TCT T markedly suppressed the IL-5Ralpha expression in the B lymphoma cellsin vitro. Ad-IL-5Ralpha could suppress the IL-5Ralpha expression of murine bone marrow cellsin vitro and it could also significantly decrease the IL-5-induced eosinophilia in the cultured bone marrow cells.
  
  
  CONCLUSIONS
  These results indicate that the blocking of IL-5Ralpha expression by small molecule inhibition can help to effectively decrease murine bone marrow eosinophilopoiesis, and that bone marrow may be used as a critical target organ in the diseases involved in eosinophilia, such as asthma.
authors.affiliation: Department of Respiratory Medicine, West China Hospital of Sichuan University, Chengdu, PR China. lfy_74@vip.163.com
authors.forename: Hui
authors.initials: H
authors.lastname: Mao
authors.forename: Fu-Qiang
authors.initials: FQ
authors.lastname: Wen
authors.forename: Su-Yun
authors.initials: SY
authors.lastname: Li
authors.forename: Zong-An
authors.initials: ZA
authors.lastname: Liang
authors.forename: Chun-Tao
authors.initials: CT
authors.lastname: Liu
authors.forename: Kai-Sheng
authors.initials: KS
authors.lastname: Yin
authors.forename: Zeng-Li
authors.initials: ZL
authors.lastname: Wang
mesh_terms: D000818:Animals; D001249:Asthma / Q000276:immunology; D001854:Bone Marrow Cells / Q000378:metabolism; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D015536:Down-Regulation; D004804:Eosinophils / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D005822:Genetic Vectors; D007249:Inflammation / Q000378:metabolism / Q000628:therapy; D053666:Interleukin-5 Receptor alpha Subunit / Q000037:antagonists & inhibitors* / Q000235:genetics; D019891:Leukopoiesis / Q000235:genetics; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D016133:Polymerase Chain Reaction; D034622:RNA Interference; D012333:RNA, Messenger / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D014162:Transfection / Q000379:methods
chemical_list: D053666:Interleukin-5 Receptor alpha Subunit; D012333:RNA, Messenger