AIM: To construct the expression vector of B3 (scdsFv)-SEA (D227A) and to identify its binding and cytotoxic ability to B3 antigen positive carcinoma cell lines.METHODS: This fusion protein was produced by a bacterial...AIM: To construct the expression vector of B3 (scdsFv)-SEA (D227A) and to identify its binding and cytotoxic ability to B3 antigen positive carcinoma cell lines.METHODS: This fusion protein was produced by a bacterial expression system in this study. It was expressed mainly in the inclusion body. The gene product was solubilized by guanidine hydrochloride, refolded by conventional dilution method, and purified using SP-sepharose cation chromatography.RESULTS: The expression vector B3 (scdsFv)-SEA-PETwas constructed, the expression product existed mainly in the inclusion body, the refolding product retained the binding ability of the single-chain antibody and had cytotoxic effect on HT-29 colon carcinoma cells. The stability assay showed that the resulting protein was stable at 37 ℃.CONCLUSION: This genetically engineered B3 (scdsFv)-SEA fusion protein has bifunction of tumor targeting and tumor cell killing and shows its promises as an effective reagent for tumor-targeted immunotherapy.展开更多
Primary biliary cirrhosis （PBC） is a progressive cho- lestatic liver disease characterized by the immune- mediated destruction of biliary epithelial cells in small intrahepatic bile ducts. The disease is characteriz...Primary biliary cirrhosis （PBC） is a progressive cho- lestatic liver disease characterized by the immune- mediated destruction of biliary epithelial cells in small intrahepatic bile ducts. The disease is characterized by circulating antimitochondrial antibodies （AMAs） as well as disease-specific antinuclear antibodies, cholestatic liver function tests, and characteristic histological fea- tures, including granulomas. A variety of organisms are involved in granuloma formation, of which mycobacte- ria are the most commonly associated. This has led to the hypothesis that mnycobacteria may be involved in the pathogenesis of PBC, along with other infectious agents. Additionally, AMAs are found in a subgroup of patients with rnycobacterial infections, such as lep- rosy and pulmonary tuberculosis. Antibodies against species-specific mycobacterial proteins have been re- ported in patients with PBC, but it is not clear whether these antibodies are specific for the disease. In addi- tion, data in support of the involvement of the role of molecular mimicry between rnycobacterial and human mitochondrial antigens as triggers of cross-reactive im- mune responses leading to the loss of immunological tolerance, and the induction of pathological features have been published. Thus, antibodies against myco- bacterial heat shock protein appear to cross-recognize AMA-specific autoantigens, but it is not clear whether these autoantibodies are mycobacterium-species-spe- cific, and whether they are pathogenic or incidental. The view that mycobacteria are infectious triggers of PBC is intriguing, but the data provided so far are not conclusive.展开更多
基金Supported by the National Natural Science Foundation of China,No. 30271478
文摘AIM: To construct the expression vector of B3 (scdsFv)-SEA (D227A) and to identify its binding and cytotoxic ability to B3 antigen positive carcinoma cell lines.METHODS: This fusion protein was produced by a bacterial expression system in this study. It was expressed mainly in the inclusion body. The gene product was solubilized by guanidine hydrochloride, refolded by conventional dilution method, and purified using SP-sepharose cation chromatography.RESULTS: The expression vector B3 (scdsFv)-SEA-PETwas constructed, the expression product existed mainly in the inclusion body, the refolding product retained the binding ability of the single-chain antibody and had cytotoxic effect on HT-29 colon carcinoma cells. The stability assay showed that the resulting protein was stable at 37 ℃.CONCLUSION: This genetically engineered B3 (scdsFv)-SEA fusion protein has bifunction of tumor targeting and tumor cell killing and shows its promises as an effective reagent for tumor-targeted immunotherapy.
文摘Primary biliary cirrhosis （PBC） is a progressive cho- lestatic liver disease characterized by the immune- mediated destruction of biliary epithelial cells in small intrahepatic bile ducts. The disease is characterized by circulating antimitochondrial antibodies （AMAs） as well as disease-specific antinuclear antibodies, cholestatic liver function tests, and characteristic histological fea- tures, including granulomas. A variety of organisms are involved in granuloma formation, of which mycobacte- ria are the most commonly associated. This has led to the hypothesis that mnycobacteria may be involved in the pathogenesis of PBC, along with other infectious agents. Additionally, AMAs are found in a subgroup of patients with rnycobacterial infections, such as lep- rosy and pulmonary tuberculosis. Antibodies against species-specific mycobacterial proteins have been re- ported in patients with PBC, but it is not clear whether these antibodies are specific for the disease. In addi- tion, data in support of the involvement of the role of molecular mimicry between rnycobacterial and human mitochondrial antigens as triggers of cross-reactive im- mune responses leading to the loss of immunological tolerance, and the induction of pathological features have been published. Thus, antibodies against myco- bacterial heat shock protein appear to cross-recognize AMA-specific autoantigens, but it is not clear whether these autoantibodies are mycobacterium-species-spe- cific, and whether they are pathogenic or incidental. The view that mycobacteria are infectious triggers of PBC is intriguing, but the data provided so far are not conclusive.
