AIM:To evaluate the predictive value of islet autoantibodies for the diagnosis of autoimmune uveitis(AU),as well as to characterize the association bet ween islet autoantibodies and AU.METHODS:Totally 97 patients with...AIM:To evaluate the predictive value of islet autoantibodies for the diagnosis of autoimmune uveitis(AU),as well as to characterize the association bet ween islet autoantibodies and AU.METHODS:Totally 97 patients with AU and 100 healthy persons without any autoimmune diseases as the control group were recruited.Multiple serum islet autoantibodies were measured using commercial enzyme-linked immunosorbent assay kits(ELISA).A supplementary questionnaire was used to complement the subject's demographics and clinical features.The level of glucose concentrations and white blood cells were measured.Conditional logistic regression was performed to estimate odds ratios(ORs),and 95%confidence intervals(CIs)of AU according to islet autoantibodies and to evaluate the predictive value of islet autoantibodies for AU diagnosis.Autoantibodies subgroups and other variables were included into analysis.RESULTS:In AU patients,the prevalence of detecting at least one of the autoantibodies was 31.9%(31/97).The most frequent autoantibody was ZnT8A(30.9%),followed by GADA(11.3%),IA-2A(4.1%),ICA(2.1%)and IAA(2.1%).Islet autoantibodies were found to be correlated positively with AU diagnosis[OR(95%CI):13.86(3.28,58.50),P<0.001].Moreover,Zn-T8A was remarkably correlated with AU diagnosis[OR(95%CI):6.13(1.96,19.17),P<0.001],In contrast,neither GADA nor other islet antibodies(IA-2A,ICA and IAA)showed any association with AU risk under an additive model.CONCLUSION:The prevalence of islet antibodies,especially ZnT8A,in patients with AU is higher.Islet antibodies as well as novel biomarkers should be included in routine evaluation at AU and is a valuable biological marker to classify newly-diagnosed uveitis.展开更多
Zinc transporter 8 （ZnT8） is a major autoantigen and a predictive marker in type 1 diabetes （T1D）. To investigate ZnT8-specific antibodies, a phage display library from TID was constructed and single-chain antibod...Zinc transporter 8 （ZnT8） is a major autoantigen and a predictive marker in type 1 diabetes （T1D）. To investigate ZnT8-specific antibodies, a phage display library from TID was constructed and single-chain antibodies against ZnT8 were screened and identified. Human T1D single-chain variable fragment （scFv） phage display library consists of approximately 1 ~ l0s clones. After four rounds of bio-panning, seven unique clones were positive by phage ELISA. Among them, C27 and C22, which demonstrated the highest affinity to ZnT8, were expressed in Escherichia coli Topl0F＇ and then purified by affin- ity chromatography. C27 and C22 specifically bound ZnT8 N/C fusion protein and ZnT8 C terminal dimer with one Arg325Trp mutation. The specificity to human islet cells of these scFvs were further confirmed by immunohistochemistry. In conclusion, we have successfully constructed a T1D phage display antibody library and identified two ZnT8-specific scFv clones, C27 and C22. These ZnT8-specific scFvs are potential agents in immunodiagnostic and immunotherapy of T1D.展开更多
基金Supported by the Science Foundation of Shaanxi Province of China(No.2017SF-265)the Science Foundation of Xi’an of China[No.2017121SF/YX015(2)].
文摘AIM:To evaluate the predictive value of islet autoantibodies for the diagnosis of autoimmune uveitis(AU),as well as to characterize the association bet ween islet autoantibodies and AU.METHODS:Totally 97 patients with AU and 100 healthy persons without any autoimmune diseases as the control group were recruited.Multiple serum islet autoantibodies were measured using commercial enzyme-linked immunosorbent assay kits(ELISA).A supplementary questionnaire was used to complement the subject's demographics and clinical features.The level of glucose concentrations and white blood cells were measured.Conditional logistic regression was performed to estimate odds ratios(ORs),and 95%confidence intervals(CIs)of AU according to islet autoantibodies and to evaluate the predictive value of islet autoantibodies for AU diagnosis.Autoantibodies subgroups and other variables were included into analysis.RESULTS:In AU patients,the prevalence of detecting at least one of the autoantibodies was 31.9%(31/97).The most frequent autoantibody was ZnT8A(30.9%),followed by GADA(11.3%),IA-2A(4.1%),ICA(2.1%)and IAA(2.1%).Islet autoantibodies were found to be correlated positively with AU diagnosis[OR(95%CI):13.86(3.28,58.50),P<0.001].Moreover,Zn-T8A was remarkably correlated with AU diagnosis[OR(95%CI):6.13(1.96,19.17),P<0.001],In contrast,neither GADA nor other islet antibodies(IA-2A,ICA and IAA)showed any association with AU risk under an additive model.CONCLUSION:The prevalence of islet antibodies,especially ZnT8A,in patients with AU is higher.Islet antibodies as well as novel biomarkers should be included in routine evaluation at AU and is a valuable biological marker to classify newly-diagnosed uveitis.
基金supported by National Natural Science Foundation of China (81070622, 81370939)Provincial Six Talent Foundation of Jiangsu (2010-022)+1 种基金Municipal Science and Technology Foundation of Nanjing (009010684)A Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD, JX10231801)
文摘Zinc transporter 8 （ZnT8） is a major autoantigen and a predictive marker in type 1 diabetes （T1D）. To investigate ZnT8-specific antibodies, a phage display library from TID was constructed and single-chain antibodies against ZnT8 were screened and identified. Human T1D single-chain variable fragment （scFv） phage display library consists of approximately 1 ~ l0s clones. After four rounds of bio-panning, seven unique clones were positive by phage ELISA. Among them, C27 and C22, which demonstrated the highest affinity to ZnT8, were expressed in Escherichia coli Topl0F＇ and then purified by affin- ity chromatography. C27 and C22 specifically bound ZnT8 N/C fusion protein and ZnT8 C terminal dimer with one Arg325Trp mutation. The specificity to human islet cells of these scFvs were further confirmed by immunohistochemistry. In conclusion, we have successfully constructed a T1D phage display antibody library and identified two ZnT8-specific scFv clones, C27 and C22. These ZnT8-specific scFvs are potential agents in immunodiagnostic and immunotherapy of T1D.
