title: [Eukaryotic expression of anti-CD3 single chain Fv antibody gene and the characterization of its bioactivities].
abstract: OBJECTIVE
  To express anti-CD3 scFv in Hela cells and investigate its biological activity.
  
  
  METHODS
  DNA fragment encoding anti-CD3 scFv was inserted into eukaryotic expression vector pDisplay. The recombinant expression vector was sequenced and then transfected into Hela cells by electroporation method. The expression of anti-CD3 scFv was identified by in situ hybridization. In-vitro T lymphocyte activation was then detected by (3)H-TdR incoporation method. Anti-CD3 scFv gene-transfected Hela cells were co-cultured with T cells and cytotoxicity was measured by MTT colorimetry.
  
  
  RESULTS
  Anti-CD3 scFv gene was correctly inserted into pDisplay and expressed in Hela cells. The secreted anti-CD3 scFv was able to activate T lymphocytes in the presence of anti-CD28 mAb. Cytotoxicity could be observed when anti-CD3 scFv gene-transfected Hela cells were mixed and co-cultured with T lymphocytes.
  
  
  CONCLUSIONS
  Anti-CD3 scFv expressed by Hela cells can activate T lymphocytes.
authors.affiliation: Institute for Cancer Research, School of Life Science & Technology, Xi'an Jiaotong University, Xi'an 710061, China. yzhangmin@yahoo.com
authors.forename: Zhang-min
authors.initials: ZM
authors.lastname: Yang
authors.forename: Jin-song
authors.initials: JS
authors.lastname: Hu
authors.forename: Bao-chang
authors.initials: BC
authors.lastname: Lai
authors.forename: Yi-li
authors.initials: YL
authors.lastname: Wang
authors.forename: Lu-sheng
authors.initials: LS
authors.lastname: Si
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D018106:CD28 Antigens / Q000276:immunology; D017252:CD3 Complex / Q000276:immunology*; D018920:Coculture Techniques; D005822:Genetic Vectors; D006367:HeLa Cells / Q000378:metabolism / Q000502:physiology; D006801:Humans; D007128:Immunoglobulin Fragments / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D008213:Lymphocyte Activation; D016853:Muromonab-CD3 / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000276:immunology*; D014162:Transfection
chemical_list: D000911:Antibodies, Monoclonal; D018106:CD28 Antigens; D017252:CD3 Complex; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D016853:Muromonab-CD3; D011993:Recombinant Fusion Proteins; C055966:immunoglobulin Fv