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Chinese Journal of Biotechnology ; (12): 703-707, 2005.
Article in Chinese | WPRIM | ID: wpr-237087

ABSTRACT

RT-PCR was used to clone DNA fragment of the extracellular domain of 4-1BBL from human THP-1 cells (human monocyte), and the expression vector pAYZ4-1BBL was constructed by cloning the extracellular domain of 4-1BBL into the expression vector pAYZ. The extracellular domain of 4-1BBL was expressed in E. coli 16C9 and purified by affinity chromatography. SDS-PAGE and Western blot analysis showed that the relativae molecular weight of soluble 4-1BBL is 22kD which was consistent with the theoretically predicted value. So far as we know, it is the first time that the soluble expression of 4-1BBL in E. coli was achieved 4-1BBL induced a significant release of IL-2 in stimulated Jurkat cells after 48h incubation, especially in the presence of tumor cell. At the same time the apoptosis level of Jurkat cell reduce more than 50%. In conclusion, 4-1BBL may be useful in cancer immunotherapy.


Subject(s)
Humans , 4-1BB Ligand , Genetics , Apoptosis , Genetics , Cell Line , Cloning, Molecular , Escherichia coli , Genetics , Metabolism , Extracellular Space , Metabolism , Interleukin-2 , Jurkat Cells , Recombinant Proteins , Genetics
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