[Effects of Schistosoma japonicum heat-shock protein 40 on macrophage activation].
OBJECTIVE
To clone and express heat-shock protein 40 gene of Schistosoma japonicum (SjHSP40) and analyze its effect on macrophage activation.


METHODS
The fragment of gene encoding SjHSP40 was amplified by PCR. The gene was sub-cloned into the prokaryotic expression vector pGEX-6P-1. The recombinant plasmid pGEX-6P-1-SjHSP40 was transformed into E. coli BL21 (DE3) and induced with IPTG. The recombinant protein was purified with Glutathione-Sepharose 4B resin and analyzed by SDS-PAGE and Western-blot. The fusion protein of GST-SjHSP40 was loaded to the macrophage cell-line RAW264.7 for 48 h. Following that, the surface molecules of the macrophages were analyzed by flow cytometry.


RESULTS
DNA sequencing showed that the recombinant plasmid, pGEX-6P-1-SjHSP40, was successfully constructed. The fusion protein of GST-SjHSP40 was induced, purified and specifically recognized by anti-GST antibody. Compared to GST and medium control groups, this fusion protein significantly induced the expression of co-stimulatory molecules (CD40, CD80, and CD86) and MHC-II on the surface of the macrophages.


CONCLUSIONS
SjHSP40 significantly up-regulates the expression of co-stimulatory molecules and MHC-II on the surface of the macrophages. These data indicate that SjHSP40 may initiate macrophage activation.
Department of Pathogenic Biology, Nanjing Medical University, Nanjing 210029, China.
Sha-Sha
SS
Li
Xiang-Ting
XT
Xu
Wei
W
Liu
Zhi-Peng
ZP
Xu
Wei-Wei
WW
Zhang
Yong
Y
Li
Xiao-Xiao
XX
Dong
Xiao-Wei
XW
Yang
Feng
F
Liu
Yue-Zhu
YZ
Wang
Sheng-Yue
SY
Wang
Chuan
C
Su
D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D050956:HSP40 Heat-Shock Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008262:Macrophage Activation / Q000276:immunology*; D008264:Macrophages / Q000276:immunology*; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012549:Schistosoma japonicum / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D012554:Schistosomiasis japonica / Q000276:immunology* / Q000378:metabolism
D050956:HSP40 Heat-Shock Proteins; D011993:Recombinant Fusion Proteins