Transgenic 4-1-BB ligand therapy induces tumor specific immune response in oral squamous cell carcinoma
10.3760/cma.j.issn.1002-0098.2009.04.002
- VernacularTitle:转人4-1-BBL口腔鳞状细胞癌体外诱导特异性抗肿瘤免疫的研究
- Author:
Shun-Tao SUN
1
;
Hong-Yu YANG
;
Juan LUO
;
Mei CHU
;
Miao ZHANG
;
Dong-Lan HUANG
Author Information
1. 北京大学深圳医院
- Keywords:
Carcinoma,squamous cell;
Gene therapy;
Cell line tumor
- From:
Chinese Journal of Stomatology
2009;44(4):198-202
- CountryChina
- Language:Chinese
-
Abstract:
Objective To examine the activation and cytotoxieity of human peripheral blood T lymphocyte induced in vitro by human 4-1-BB ligand(4-1-BBL) gene transfected into tumor Tca8113 cells.Methods The eukaryotic expression vector pEGFP-h4-1-BBL was transfected into human oral carcinoma cell line Tca8113 by Lipofectamine<'TM>2000. The transfected cells were then selected in medium containing G-418, cloned by limited dilution and named as Tca8113-4-1-BBL Human 4-1-BBL mRNA and protein expression of transfected cells was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) and western blotting respectively. The tumor cell vaccines (TCV) were obtained by treatment with mitomycin (MMC). Human peripheral blood mononuclenr cells (PBMC) were prepared from lymphoprep, and then stimulated with anti-CD-3 mAb and incubated with non-transfected or transfected TCV-TcaSI13 cells,respectively. The proliferation of T cells was evaluated by trypan blue exclusion; the CCK-8 was used to detect the cytotoxic effect of T lymphecytes. Meanwhile, the secretion of intefferom-γ (IFN-γ) and interleukin(IL)-2 in culture supernatant was detected by enzyme-linked immunosorbnent assay (ELISA).Results The Tca8113 cells trasfected by pEGFP-h4-1-BBL could express human 4-I-BBL efficiently. As compared with wild type Tca8113 cells, the transfected Tca8113 cells could markedly promote proliferation,IL-2 and IFN-γ production and cytotoxic activity of lymphocytos. Condusions The transfection of human 4-1-BBL gene in Tca8113 cells is effctive in enhancing its immunogenicity and inducing antitumor immune response in vitro.
