Molecular cloning and expression of a functional anti-inflammatory protein, Sj16, of Schistosoma japonicum.
Schistosomes are the causative agent of schistosomiasis. In the infected host, significant inflammatory response to the parasite is not observed. Previous studies of Schistosoma mansoni showed that this subdued inflammatory response was due to a 16-kDa protein, Sm16, which is present in high levels in the secretions of schistosomula. Here we report the cloning and characterization of a gene (named Sj16) from Schistosoma japonicum. Sequence analysis showed that Sj16 shares 99% identity with Sm16 in its nucleotide sequence, and 100% identity in its protein sequence. While previous studies reportedly failed to obtain the soluble recombinant protein of Sm16, we expressed and purified recombinant Sj16 (rSj16) from Escherichia coli. Western blot and ELISA analyses showed that S. japonicum-infected rabbit sera could not recognize rSj16, indicating that native Sj16 may fail to induce circulating antibodies during S. japonicum infection. In vivo, rSj16 dramatically suppressed the recruitment of thioglycollate-mediated leukocytes to the peritoneal cavity of BALB/c mice, accompanied by marked up-regulation of IL-10 and IL-1RA transcripts, and down-regulation of IL-12p35, IL-1 beta and MIP-2 transcripts in peritoneal cells. Further analysis revealed that rSj16 also suppressed thioglycollate-induced peritoneal macrophage maturation. These results demonstrate that rSj16 has an anti-inflammatory function.
Hu
Shaomin
S
Department of Biology, The Chinese University of Hong Kong, Shatin, Hong Kong, China.
Wu
Zhongdao
Z
Yang
Linlin
L
Fung
Ming Chiu
MC
D000818:Animals; D000909:Antibodies, Helminth / Q000378:metabolism; D003001:Cloning, Molecular; D016207:Cytokines / Q000378:metabolism*; D004195:Disease Models, Animal; D015536:Down-Regulation; D004926:Escherichia coli / Q000235:genetics* / Q000276:immunology; D005260:Female; D015801:Helminth Proteins / Q000235:genetics / Q000502:physiology*; D007249:Inflammation / Q000276:immunology; D007962:Leukocytes / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D010529:Peritoneal Cavity / Q000166:cytology; D011817:Rabbits; D012549:Schistosoma japonicum / Q000235:genetics* / Q000276:immunology; D013864:Thioglycolates / Q000378:metabolism; D015854:Up-Regulation
D000909:Antibodies, Helminth; D016207:Cytokines; D015801:Helminth Proteins; D013864:Thioglycolates