[Eukaryotic expression of anti-CD3 single chain Fv antibody gene and the characterization of its bioactivities].
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.
Yang
Zhang-min
ZM
Institute for Cancer Research, School of Life Science & Technology, Xi'an Jiaotong University, Xi'an 710061, China. yzhangmin@yahoo.com
Hu
Jin-song
JS
Lai
Bao-chang
BC
Wang
Yi-li
YL
Si
Lu-sheng
LS
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
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