An Analysis of the Cross-Reactivity of Autoantibodies to GAD65 and GAD67 in Diabetes 英文参考文献.docVIP

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An Analysis of the Cross-Reactivity of Autoantibodies to GAD65 and GAD67 in Diabetes 英文参考文献.doc

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An Analysis of the Cross-Reactivity of Autoantibodies to GAD65 and GAD67 in Diabetes 英文参考文献

AnAnalysisoftheCross-ReactivityofAutoantibodiesto GAD65andGAD67inDiabetes BinduJayakrishnan,DavidE.Hoke,ChristopherG.Langendorf,AshleyM.Buckle*.,MerrillJ.Rowley*. DepartmentofBiochemistryandMolecularBiology,MonashUniversity,Clayton,Victoria,Australia Abstract Background: AutoantibodiestoGAD65(anti-GAD65)arepresentintheseraof70–80%ofpatientswithtype1diabetes (T1D),butantibodiestothestructurallysimilar67kDaisoformGAD67arerare.AntibodiestoGAD67mayrepresentacross- reactivepopulationofanti-GAD65,butthishasnotbeenformallytested. Methodology/PrincipalFindings:Inthisstudyweexaminedthefrequency,levelsandaffinityofanti-GAD67indiabetessera thatcontainedanti-GAD65,andcomparedthespecificityofGAD65andGAD67reactivity.Anti-GAD65andanti-GAD67were measured by radioimmunoprecipitation (RIP) using 125I labeled recombinant GAD65 and GAD67. For each antibody population,thespecificityofthebindingwasmeasuredbyincubationwith100-foldexcessofunlabeledGADinhomologous andheterologousinhibitionassays,andtheaffinityofbindingwithGAD65andGAD67wasmeasuredinselectedsera.Sera werealsotestedforreactivitytoGAD65andGAD67byimmunoblotting.Ofthe85serathatcontainedantibodiestoGAD65,28 contained anti–GAD67 measured by RIP. Inhibition with unlabeled GAD65 substantially or completely reduced antibody reactivitywithboth125IGAD65andwith125IGAD67.Incontrast,unlabeledGAD67reducedautoantibodyreactivitywith 125I GAD67butnotwith 125IGAD65.Bothpopulationsofantibodieswereofhighaffinity(.1010 l/mol). Conclusions:OurfindingsshowthatautoantibodiestoGAD67representaminorpopulationofanti-GAD65thatarereactive withacross-reactiveepitopefoundalsoonGAD67.ExperimentalresultsconfirmthatGAD65isthemajorautoantigenin T1D,andthatGAD67persehasverylowimmunogenicity.Wediscussourfindingsinlightoftheknownsimilaritiesbetween thestructuresoftheGADisoforms,inparticularthelocationofaminorcross-reactiveepitopethatcouldbeinducedby epitopespreading. Citation:JayakrishnanB,HokeDE,LangendorfCG,BuckleAM,RowleyMJ(2011)AnAnalysisoftheCross