[Construction of anti-cD20scFv/CD80/CD28/zeta recombinant gene modified T cell and research on its targeting cytotoxicity].
OBJECTIVE
To construct anti-CD20scFv/CD80/CD28/zeta recombinant gene modified T cells, test its effectiveness of eradicating CD20+ lymphoma cells and provide a probably new approach to tumor adoptive immunotherapy.


METHODS
CD28-zeta cDNA were amplified from vector pBULLET and inserted into pLNCX vector that contained anti-CD20scFv/CD80 gene. The recombinant vectors were transduced into PA317 cells and high titer retroviruses were obtained to infect human peripheral blood T lymphocytes. Resistant T cells were obtained by G418 selection at one week. Then transduced T lymphocytes and lymphoma cell lines Daudi Raji were cocultured. The cytotoxicity and cytokine production of transduced T cells were determined by non-radio-activation cytotoxicity assay and ELISA respectively.


RESULTS
The recombinant eukaryotic vector was constructed successfully as proved by enzyme digestion analysis and sequencing. These T cells were able to lyse CD20+ target cells and secrete high levels of IL-2 and IFN-gamma in vitro.


CONCLUSIONS
Recombinant gene modified T cells can be constructed successfully. It can specially kill CD20 positive lymphoma cells in vitro.
Hu
Yong-Xian
YX
The First Affiliated Hospital, Wenzhou Medical College, Wenzhou 325000, China.
Yu
Kang
K
Tan
Ying-Xia
YX
Shen
Zhi-Jian
ZJ
Jiang
Song-Fu
SF
Qian
Hong-Lan
HL
Hang
Bin
B
Shan
Da-Ming
DM
D018951:Antigens, CD20 / Q000235:genetics / Q000276:immunology*; D018122:B7-1 Antigen / Q000235:genetics / Q000276:immunology*; D018106:CD28 Antigens / Q000235:genetics* / Q000276:immunology; D002460:Cell Line; D003602:Cytotoxicity, Immunologic; D005822:Genetic Vectors; D006801:Humans; D016219:Immunotherapy, Adoptive; D010957:Plasmids / Q000235:genetics; D013601:T-Lymphocytes / Q000276:immunology* / Q000378:metabolism; D014162:Transfection
D018951:Antigens, CD20; D018122:B7-1 Antigen; D018106:CD28 Antigens