[Cloning and expression of the extracellular domain of 4-1BBL].
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.
Jiang
Wen-Guo
WG
State Key Laboratory of Experimental Hematology , Institute of Hematology, Chinese Academy of Medical Science & Peking Union Medical College, Tianjin 300020, China.
Xiong
Dong-Sheng
DS
Shao
Xiao-Feng
XF
Wang
Jin-Hong
JH
Xu
Yuan-Fu
YF
Liu
Fang
F
Guo
Hong-Xing
HX
Zhu
Zhen-Ping
ZP
Yang
Chun-Zheng
CZ
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
D053330:4-1BB Ligand; D007376:Interleukin-2; D011994:Recombinant Proteins