title: [Eukaryotic expression of recombinant B7-H3Ig protein and its  effects on T cells].
abstract: OBJECTIVE
  To construct a recombinant eukaryotic expression vector containing the human B7-H3Ig gene and investigate the effects of B7-H3 signal on T cells.
  
  
  METHODS
  Being constructed by overlap extension PCR, the recombinant gene fragment of B7-H3Ig was inserted into pGEZ-Term vector. The recombinant pGEZ-Term/B7-H3Ig vector was used to transfect L929 cells to purify B7-H3Ig from conditioned medium harvested from cultured L929/B7-H3Ig cells. T cells were then stimulated with agonistic anti-CD3 mAb in the presence or absence of purified B7-H3Ig, and cell proliferation and the secretion of IL-10 and IFN-γ were analyzed 72 h later.
  
  
  RESULTS
  The concentration of B7-H3Ig was 0.559 mg/mL with a purity of more than 90% after protein G affinity chromatography. B7-H3Ig binding test showed that the putative B7-H3 receptor was presented in activated T cells and the maximum binding capacity was observed 48 h later. B7-H3Ig enhanced, in a dose-dependent manner, the proliferation and IL-10 and IFN-γ secretion of T cells stimulated with agonistic anti-CD3 mAb.
  
  
  CONCLUSIONS
  L929/B7-H3Ig transfectants have been successfully constructed. B7-H3 is a co-stimulator for T cell proliferation and IL-10 and IFN-γ expressions.
authors.affiliation: Department of Clinical Laboratory, Second Affiliated Hospital, Anhui Medical University, Hefei 230601, China. lw_ch@163.com
authors.forename: Liwen
authors.initials: L
authors.lastname: Chen
authors.forename: Guangbo
authors.initials: G
authors.lastname: Zhang
authors.forename: Yibei
authors.initials: Y
authors.lastname: Zhu
authors.forename: Xueguang
authors.initials: X
authors.lastname: Zhang
mesh_terms: D060887:B7 Antigens / Q000235:genetics* / Q000276:immunology / Q000494:pharmacology; D002460:Cell Line; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D016753:Interleukin-10 / Q000378:metabolism; D008213:Lymphocyte Activation / Q000276:immunology; D011994:Recombinant Proteins / Q000235:genetics* / Q000276:immunology / Q000494:pharmacology; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism*
chemical_list: D060887:B7 Antigens; C423237:CD276 protein, human; D011994:Recombinant Proteins; D016753:Interleukin-10; D007371:Interferon-gamma