Construction of anti-cD20scFv/CD80/CD28/zeta recombinant gene modified T cell and research on its targeting cytotoxicity.
- Author:
Yong-Xian HU
1
;
Kang YU
;
Ying-Xia TAN
;
Zhi-Jian SHEN
;
Song-Fu JIANG
;
Hong-Lan QIAN
;
Bin HANG
;
Da-Ming SHAN
Author Information
- Publication Type:Journal Article
- MeSH: Antigens, CD20; genetics; immunology; B7-1 Antigen; genetics; immunology; CD28 Antigens; genetics; immunology; Cell Line; Cytotoxicity, Immunologic; Genetic Vectors; Humans; Immunotherapy, Adoptive; Plasmids; genetics; T-Lymphocytes; immunology; metabolism; Transfection
- From: Chinese Journal of Hematology 2007;28(2):111-114
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSCD28-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.
RESULTSThe 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.
CONCLUSIONRecombinant gene modified T cells can be constructed successfully. It can specially kill CD20 positive lymphoma cells in vitro.
