Abstract
Schistosomiasis is one of the most prevalent parasitic diseases in China, and hepatic fibrosis caused by schistosome infection is the principal cause of death. The aim of this study was to evaluate the efficacy of NP11-4-derived immunotoxin scFv-artesunate on Schistosoma japonicum-induced hepatic fibrosis. A single-chain variable fragment (scFv) was generated from the murine anti- Schistosoma japonicum (S. japanicum) monoclonal antibody NP11-4. The scFv was expressed as a soluble protein and purified by Ni-affinity chromatography. After conjugation with artesunate, the binding ability with soluble egg antigens (SEA) was determined by an enzyme-linked immunosorbent assay (ELISA). The biological activity of purified scFv, scFv-artesunate (immunotoxin), and artesunate was detected in vivo. Image-Pro Plus software was used to analyze the size of egg granuloma and the extent of liver fibrosis. The recombinant scFv expession vector was constructed and expressed successfully. After purification by a His-trap Ni-affinity column, the scFv yield was approximately 0.8 mg/L of culture medium. ELISA results showed that chemical conjugation did not affect the binding activity of the immunotoxin. Our animal experiments indicated that the immunotoxin could significantly reduce the size of egg granuloma in the liver and inhibit hepatic fibrosis. The immunotoxin could be used as a promising candidate in the targeted therapy of S. japonicum-induced hepatic fibrosis.
Highlights
Schistosomiasis, which is caused by several species of the trematode Schistosoma, is an endemic disease in 74 developing countries
The variable heavy (VH) and variable light (VL) genes were amplified from the mRNA, which was isolated from the parental hybridoma cell line NP11-4
Using the primers VHF and VLR, the scFv gene was amplified by overlap PCR
Summary
Schistosomiasis, which is caused by several species of the trematode Schistosoma, is an endemic disease in 74 developing countries. In individuals infected with S. japonicum, chronic egg-induced inflammation in the periportal tracts of the liver can lead to fibrosis, portal hypertension, bleeding, and eventually death[1,2,3,4]. In China, schistosomiasis japonica is endemic in the 12 provinces along the southern part of the Yangtze River[5]. The estimated number of cases of chronic. Efficacy of immunotoxin in hepatic fibrosis schistosomiasis japonica in 2008 is 412,927, which represents a great reduction from the 690,000 reported cases for 2000[6,7,8]. Re-infection with schistosomes can occur even after successful chemotherapy[9]
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