title: [Cloning and expression of the extracellular domain of 4-1BBL].
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.
authors.affiliation: State Key Laboratory of Experimental Hematology , Institute of Hematology, Chinese Academy of Medical Science & Peking Union Medical College, Tianjin 300020, China.
authors.forename: Wen-Guo
authors.initials: WG
authors.lastname: Jiang
authors.forename: Dong-Sheng
authors.initials: DS
authors.lastname: Xiong
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Shao
authors.forename: Jin-Hong
authors.initials: JH
authors.lastname: Wang
authors.forename: Yuan-Fu
authors.initials: YF
authors.lastname: Xu
authors.forename: Fang
authors.initials: F
authors.lastname: Liu
authors.forename: Hong-Xing
authors.initials: HX
authors.lastname: Guo
authors.forename: Zhen-Ping
authors.initials: ZP
authors.lastname: Zhu
authors.forename: Chun-Zheng
authors.initials: CZ
authors.lastname: Yang
mesh_terms: D053330:4-1BB Ligand / Q000096:biosynthesis* / Q000235:genetics*; D017209:Apoptosis / Q000235:genetics; D002460:Cell Line; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005110:Extracellular Space / Q000378:metabolism; D006801:Humans; D007376:Interleukin-2 / Q000096:biosynthesis; D019169:Jurkat Cells; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D053330:4-1BB Ligand; D007376:Interleukin-2; D011994:Recombinant Proteins