title: [Eukaryotic expression and biological activity of recombinant human IL-17B protein].
abstract: OBJECTIVE
  To construct pCEP4/hIL-17B recombinant expression vector and express it stably in eukaryotic cells and investigate the biological activity in vitro.
  
  
  METHODS
  The CDS region of human IL-17B gene was cloned by RT-PCR. After identification by sequencing,  the hIL-17B gene was inserted into expression vector of pCEP4 to construct the recombinant vector pCEP4/hIL-17B,  then transfected into 293T cells. The transgenic 293T cell line stably expressing rhIL-17B protein was selected in the presence of Hygromycin B. After FCS-free cultivation and sub-cloning,  The IL-17B/mFc gene and protein expression was confirmed by RT-PCR,  ELISA and Western blot analysis. To investigate the ability of combination with IL-17B receptor on human leukemic monocytic cell line,  THP-1,  by Flow cytometrical analysis (FACS) and of stimulation to secret cytokines in vitro.
  
  
  RESULTS
  The recombinant pCEP4/hIL-17B and its transgenic 293T cells stably expressing rhIL-17B protein were obtained successfully. FACS analysis showed its high affinity with its receptor and it can stimulated THP-1 cell line to excrete IL-1β and TNF-α in vitro and consistently caused a dose-dependent influx of neutrophil into the peritoneal cavity by intraperitoneal injection in vivo.
  
  
  CONCLUSIONS
  The obtainment of transgenic 293T cell line stably expressing rhIL-17B protein paved the way for further study on biological functions of hIL-17B.
authors.affiliation: Department of Ophthalmology, The First Affiliated Hospital of Soochow University, Suzhou, China. liugaoqin2006@sina.com
authors.forename: Gao-qin
authors.initials: GQ
authors.lastname: Liu
authors.forename: Long-biao
authors.initials: LB
authors.lastname: Li
authors.forename: Song-guang
authors.initials: SG
authors.lastname: Ju
authors.forename: Wen-peng
authors.initials: WP
authors.lastname: Zhang
authors.forename: Yi-yong
authors.initials: YY
authors.lastname: Qian
authors.forename: Xue-fei
authors.initials: XF
authors.lastname: Zhu
authors.forename: Pei-rong
authors.initials: PR
authors.lastname: Lu
authors.forename: Xue-guang
authors.initials: XG
authors.lastname: Zhang
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D020381:Interleukin-17 / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D051379:Mice; D010957:Plasmids / Q000235:genetics; D053722:Receptors, Interleukin-17 / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: D018076:DNA, Complementary; D020381:Interleukin-17; D053722:Receptors, Interleukin-17; D011994:Recombinant Proteins