Schistosoma japonicum scFv-IL18 fusion DNA ameliorates hepatic fibrosis in schistosomiasis-infected mice via improving local concentration of IL-18 in liver.

The pathogenesis of chronic schistosomiasis is caused by irritation of the schistosome eggs trapped in liver that induce delayed hypersensitive reactions from the surrounding tissues, leading to the formation of inflammatory granuloma and subsequent fibrosis. A Schistosoma japonicum (S. japonicum) single-chain fragment variable (SjscFv) which specifically binds to the S. japonicum soluble immature egg antigen (SIEA) can be used as a target to deliver specific cytokine towards the site of hepatic fibrosis. To test this hypothesis, a novel recombinant plasmid, pVAX1/SjscFv-IL18, was constructed by fusing SjscFv to IL-18 gene with a 45bp glycine-rich linker. Furthermore, experiments on mice showed that pVAX1/SjscFv-IL18 could effectively express IL-18 in the liver and in serum. Hepatic contents of IL-2 and IFN-γ (Th1-type) in S. japonicum-infected mice vaccinated with pVAX1/SjscFv-IL18 increased significantly but those of their IL-4 and IL-10 (Th2-type) decreased as compared to the analyzed results of 4 cytokines in the liver cells of control mice vccinated with pVAX1/IL18. Consistent with the levels of Th1 and Th2 cytokines, mice vaccinated with pVAX1/SjscFv-IL18 developed much less hepatic fibrosis 20weeks after infection, which was evaluated by average volumn of granuloma and collagen contents. These data suggested that the linkage of IL-18 to the target-specific SjscFv molecule appears to be a potentially promising trial route of therapy, the hepatic fibrosis in S. japonicum-infected mice may be ameliorated through effective expression of IL18 in liver.

Characterization and vaccination of two novel Schistosoma japonicum genes screened from a cercaria cDNA library.

Two novel genes, SJCWL05 and SJCWL06, were harvested from screening of Schistosoma japonicum (S. japonicum) cercaria cDNA library by using pig sera vaccinated (VPS) with S. japonicum immature egg ws-vaccine (S. japonicum iEw). Prokaryotic recombinant plasmids pGEX-4T-1/SJCWL05 and pGEX-4T-1/SJCWL06 were constructed to analyze their immunogenicity, which was confirmed by SDS-PAGE and Western blotting. Two eukaryotic recombinant plasmids, pcDNA3/SJCWL05 and pcDNA3/SJCWL06, were constructed, and their ability to protect mice against challenge of S. japonicum was evaluated. All mice vaccinated with pcDNA3/SJCWL05 or pcDNA3/SJCWL06 developed ELISA-specific anti-S. japonicum SIEA (S. japonicum soluble immature egg antigens) antibody. Immunoprotection experiments showed that worms and liver eggs reduced 34.64% and 39.14% in the pcDNA3/SJCWL05 group and those reduced 27.17% and 27.95% in the pcDNA3/SJCWL06 group, respectively. The reduction rates of intestine and uterine eggs in female worms of both groups reached 39.45% and 38.5% as well as 30.02% and 28.7%, respectively. Results of our study suggest that novel genes, SJCWL05 and SJCWL06, are potential vaccine candidates against schistosomiasis japonica.