Expression of human IL-12 in mammalian cell and study on its biological activities.
- Author:
Hong-Yuan JIAO
1
;
Mei-Yun ZHAN
;
Min-Zhuo GUO
;
Yao YI
;
Yu CONG
;
Rui-Guang TIAN
;
Wen-Ying ZHANG
;
Sheng-Li BI
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Blotting, Western; CHO Cells; Cell Proliferation; drug effects; Cricetinae; Cricetulus; Dose-Response Relationship, Drug; Genetic Vectors; genetics; Humans; Interleukin-12; biosynthesis; genetics; pharmacology; Polymerase Chain Reaction; Recombinant Proteins; biosynthesis; metabolism; pharmacology; T-Lymphocytes; cytology; drug effects; Transfection
- From: Chinese Journal of Experimental and Clinical Virology 2007;21(3):235-237
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo obtain recombinant human interleukin 12 by genetic engineering, and to explore possibility of its clinical application in treatment of tumor and chronic hepatitis.
METHODSBicistronic expression vector P35-IRES-P40 was constructed for the simultaneous translation of IL-12 p35 and p40 cDNA subunit through internal ribosomal entry sites (IRES). pCI-dhfr-P35-IRES-P40 vector was constructed for expression in CHO-DHFR- cells. Positively cloned cells were screened by means of ELISA. Pools of clones with increased expression of IL-12 could be generated by selection in methotrexate. To determine the biological activities of rhIL-12, PHA-activated lymphoblasts proliferation assay and IFN-gamma induction assay were used in this study.
RESULTSGenetically engineered cells expressing hIl-12 were obtained and all the cell lines showed the stabile expression of rhIL-12 in high efficiency and good growth properties.
CONCLUSIONrhIL-12 have good biological activities, it can stimulate activation and proliferation of T cells and induce production of IFN-gamma.
