title: [Preliminary identification of T cell epitopes on 22.6 kDa antigen of Schistosoma japonicum].
abstract: OBJECTIVE
  To identify the T cell epitopes on 22.6 kDa antigen of Schistosoma japonicum (Sj22.6).
  
  
  METHODS
  The primary structure of Sj22.6 molecule was analysed using various predictive algorithms and a panel of 4 peptides were acquired. Their oligonucleotides were designed, synthesized and inserted into the multiple cloning site of plasmid pET-32c(+). The recombinant plasmids were transformed into E. coli BL21 and identified by endonuclease digestion and sequencing. The positive clones containing the recombinant plasmids could express specific fusion proteins (trx-epitope, MW approximately 20 kDa) induced by IPTG. The fusion protein with 6 x His could be coupled with NTA resin specifically, and purified by elusion of the column with buffer containing imidazole. The purified fusion proteins were incubated with splenocytes of C3H mice and then, the proliferation of splenocytes was determined by 3H-TdR incorporation assay.
  
  
  RESULTS
  The recombinant plasmids were constructed successfully and the positive clones containing the recombinant plasmids expressed specific fusion proteins. Three of the purified fusion proteins (P4, P5, P6) could stimulate the lymphocyte proliferation.
  
  
  CONCLUSIONS
  Three T cell epitopes on Sj22.6 antigen were identified.
authors.affiliation: Institute for Molecular Biology of Medicine, Nanjing Medical University, Nanjing 210029.
authors.forename: Xin-jun
authors.initials: XJ
authors.lastname: Wang
authors.forename: Zhao-song
authors.initials: ZS
authors.lastname: Zhang
authors.forename: Guang-fu
authors.initials: GF
authors.lastname: Li
authors.forename: Yong
authors.initials: Y
authors.lastname: Wang
authors.forename: Feng
authors.initials: F
authors.lastname: Liu
authors.forename: Min-jun
authors.initials: MJ
authors.lastname: Ji
authors.forename: Xiang
authors.initials: X
authors.lastname: Zhu
authors.forename: Xiao-ping
authors.initials: XP
authors.lastname: Cai
authors.forename: Guan-ling
authors.initials: GL
authors.lastname: Wu
mesh_terms: D000818:Animals; D000947:Antigens, Helminth / Q000096:biosynthesis* / Q000276:immunology; D002478:Cells, Cultured; D000939:Epitopes; D005260:Female; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000276:immunology; D051379:Mice; D008809:Mice, Inbred C3H; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000276:immunology; D012549:Schistosoma japonicum / Q000276:immunology*; D013154:Spleen / Q000166:cytology
chemical_list: C087430:22.6-kDa antigen, Schistosoma japonicum; D000947:Antigens, Helminth; D000939:Epitopes; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins