title: [Construction of human CXCR3B gene transfected cell line,   identification and study of its biological function].
abstract: OBJECTIVE
  To construct recombinant human CXCR3B gene expression vector and obtain L929-CXCR3B gene transfected cell line for stably expressing human CXCR3B.
  
  
  METHODS
  Human CXCR3B gene of full length was amplified by PCR from the plasmid pMD19-T/huCXCR3A. Then,   it was inserted into eukaryotic expression vector pIRES2-EGFP to construct recombinant vector pIRES2-EGFP/huCXCR3B. The recombinant vector was transfected into L929 cells with Lipofectamine(TM); 2000,  and cell clones stably expressing human CXCR3B molecule were screened by G418.We used FCM and RT-PCR to verify expression of CXCR3B from protein level and gene level. The ability of proliferation of L929-huCXCR3B under the circumstance of CXCR3B ligand called Mig was analyzed via MTT methods.
  
  
  RESULTS
  We have constructed recombinant human CXCR3B gene expression vector and obtained L929-huCXCR3B gene transfected cell line which can stably express human CXCR3B molecule. The positive expression rate of CXCR3B on L929-huCXCR3B cells was 93&.  The  result of MTT assay showed  that the proliferation of L929-huCXCR3B cells were inhibited when the cells were cocultured with Mig after 24 h,  48 h and 72 h,  and the inhibition ratio were 41.44&,  44.01& and 24.80& respectively.
  
  
  CONCLUSIONS
  Construction of L929-huCXCR3B cell line has laid a good foundation on research of the CXCR3B signal pathway and preparation of the CXCR3B monoclonal antibody.
authors.affiliation: Department of Immunology, Medical College of Soochow University, Suzhou 215123, China. guojingya1118@163.com
authors.forename: Jing-ya
authors.initials: JY
authors.lastname: Guo
authors.forename: Chang-you
authors.initials: CY
authors.lastname: Chen
authors.forename: Zhi-qiang
authors.initials: ZQ
authors.lastname: Wang
authors.forename: Yan-jun
authors.initials: YJ
authors.lastname: Zhang
authors.forename: Sai-nan
authors.initials: SN
authors.lastname: Huang
authors.forename: Hua-ting
authors.initials: HT
authors.lastname: Zhu
authors.forename: Yu-hua
authors.initials: YH
authors.lastname: Qiu
mesh_terms: D002460:Cell Line; D049109:Cell Proliferation; D054370:Chemokine CXCL9 / Q000494:pharmacology; D003001:Cloning, Molecular; D005434:Flow Cytometry; D005822:Genetic Vectors; D006801:Humans; D019718:Receptors, CXCR4 / Q000235:genetics* / Q000502:physiology; D014162:Transfection
chemical_list: C080810:CXCL9 protein, human; C527276:CXCR4 protein, human; D054370:Chemokine CXCL9; D019718:Receptors, CXCR4