Specific activity of cytotoxic T lymphocyte in mice induced by dendritic cells after mIL-12 gene modification and murine colon carcinoma RNA transfection
- VernacularTitle:小鼠肠癌RNA转染mIL-12修饰的树突细胞诱发特异性细胞毒性T淋巴细胞的研究
- Author:
Xiaoyuan CHU
- Publication Type:Journal Article
- Keywords:
Interleukin 12;
Gene therapy;
Dendritic cells;
Cytotoxic T lymphocyte
- From:
Journal of Medical Postgraduates
2004;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate specific activity of cytotoxic T lymphocyte(CTL) induced by(mIL-12) gene modified dendritic cells(DCs) transfected with the total RNA of CT-26(a cell line of murine carcinoma of colon).Methods:In vitro proliferation of DCs from the culture of murine bone marrow was stimulated by rmGM-CSF and rmIL-4,and the purity of DCs was detected by flow-cytometry.The generated DCs were then modified by adenovirus with mIL-12 gene which were proliferated in 293 cells.The total RNA of CT-26 was obtained through Trizol's process,and transfected into the mIL-12 gene modified DC by TransMessenger in vitro.The levels of mIL-12 both in vitro and in vivo and the activity of CTL in vivo were estimated with enzyme linked immunosorbent assay and modified lactate dehydrogenase release assay respectively.Results:Plenty of DCs were obtained from the culture of murine bone marrow,with over 90 % of CD11c~(+) cells.DC modified by mIL-12 gene could induce high level of mIL-12 both in vitro and in vivo,and the differences compared to control groups were significant(P