[Preliminary identification of T cell epitopes on 22.6 kDa antigen of Schistosoma japonicum].
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.
Wang
Xin-jun
XJ
Institute for Molecular Biology of Medicine, Nanjing Medical University, Nanjing 210029.
Zhang
Zhao-song
ZS
Li
Guang-fu
GF
Wang
Yong
Y
Liu
Feng
F
Ji
Min-jun
MJ
Zhu
Xiang
X
Cai
Xiao-ping
XP
Wu
Guan-ling
GL
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
C087430:22.6-kDa antigen, Schistosoma japonicum; D000947:Antigens, Helminth; D000939:Epitopes; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins