title: [Establishment of the transfected cell line expressing the mouse OX40 gene and its potency to induce differentiation of B cells].
abstract: OBJECTIVE
  To establish HUVECs line expressing mouse OX40(CD134) and to study its promotive effect on proliferation of B cells.
  
  
  METHODS
  The cDNA fragment encoding mouse OX40 was obtained from the total RNA of ConA-activated lymphocytes of thymus by using RT-PCR and cloned into pUCm-T vector. The cDNA was then inserted into the eukaryotic expression vector pIRES2-EGFP. The recombinant vector was transfected into HUVECs with lipofectin reagent, and the positive cellular clones were selected by G418. Expression of mouse OX40 in the transfected cells was analyzed by FCM. The differentiation of B cells in vitro induced by OX40 signal was studied by means of (3)H-TdR method.
  
  
  RESULTS
  The cDNA fragment in the recombinant plasmid was consistent with the reported mouse OX40 cDNA in GenBank, which was confirmed by DNA sequencing, PCR and enzyme digestion. The HUVECs stably expressing the mouse OX40 were successfully cloned. The transfected cells promoted the differentiation of B cells in vitro and there existed a synergic effect between OX40/OX40L and CD40/CD40L signals.
  
  
  CONCLUSIONS
  Transfected cell line expressing the mouse OX40 gene is established successfully. OX40 enhances the proliferation of B cells.
authors.affiliation: Laboratory of Molecular Immunology, Weifang Medical College, Weifang 261042, China.
authors.forename: Jiang-shui
authors.initials: JS
authors.lastname: Yuan
authors.forename: Yong-tang
authors.initials: YT
authors.lastname: Feng
authors.forename: Fei
authors.initials: F
authors.lastname: Wang
authors.forename: Wei-yan
authors.initials: WY
authors.lastname: Gong
authors.forename: Da-lin
authors.initials: DL
authors.lastname: Di
authors.forename: Jie
authors.initials: J
authors.lastname: Ren
authors.forename: Nai-fa
authors.initials: NF
authors.lastname: Miao
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology* / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D002460:Cell Line; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D053262:Receptors, OX40 / Q000235:genetics / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection / Q000379:methods
chemical_list: D018076:DNA, Complementary; D053262:Receptors, OX40; C505717:Tnfrsf4 protein, mouse