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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 NP11-4derived 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.
Schistosomiasis is one of the most prevalent parasitic diseases in China, and hepatic fibrosis caused by schistosome infection is the principal cause of death of. 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. japanum) 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. recombinant scFv expession vector was constructed and expressed successfully. After purification by a His-tra p 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 that 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.