title: [Preparation of anti-human 4-1BB monoclonal antibody and characterization of its biological activities].
abstract: OBJECTIVE
  To prepare an anti-human 4-1BB functional monoclonal antibody and to characterize its biological activities.
  
  
  METHODS
  A stable human 4-1BB molecule transfected cell line 293T/4-1BB was used as an antigen to immunize BALB/c mice. By means of the cell fusion by hybridoma technique and multiple cell subcloning and repeated screening with 293T/4-1BB as the antibody screening positive cell while 293T/mock as the negative cell. The hybridoma cell lines specifically secreting anti-4-1BB monoclonal antibodies were selected. Then their characteristics and its biological activities were investigated by Western blot, fast-strip routine Ig subclass typing method, indirect immunofluorescence, competitive inhibition test, (3);H-TdR and cell apoptosis analysis.
  
  
  RESULTS
  Three hybridoma cell lines 1G5, 4B11 and 9F11 with the property of secreting specific anti-4-1BB monoclonal antibody continuously and steadily were successfully obtained. These monoclonal antibodies could bind to human 4-1BB epitopes on activated T cells and monocytoes and DC. Additionally, mAb 4B11 could promote T proliferation and enhance the growth and maturation of Mo-DC.
  
  
  CONCLUSIONS
  Three hybridoma cell lines which secrete anti-4-1BB monoclonal antibodies steadily have been established. These monoclonal antibodies could specifically recognize 4-1BB molecule and mAb 4B11 had a potent function to promote T proliferation cell as well as to enhance the growth and maturation of Mo-DC in vitro.
authors: Department of Clinical Laboratory, No.4 Affiliated Hospital of Suzhou University, Wuxi 214062, China.
authors: Huan
authors: H
authors: Zhou
authors: Guang-Bo
authors: GB
authors: Zhang
authors: Qiu-Ming
authors: QM
authors: Dong
authors: Ge-Hua
authors: GH
authors: Yu
authors: Ying
authors: Y
authors: Xu
authors: Xue-Guang
authors: XG
authors: Zhang
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification*; D000918:Antibody Specificity / Q000276:immunology; D003713:Dendritic Cells / Q000276:immunology; D000939:Epitopes / Q000276:immunology; D057809:HEK293 Cells; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D008213:Lymphocyte Activation / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D013601:T-Lymphocytes / Q000276:immunology; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9 / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000939:Epitopes; C505714:TNFRSF9 protein, human; D053261:Tumor Necrosis Factor Receptor Superfamily, Member 9