韦格纳肉芽肿病自身抗原蛋白酶3的抗原决定簇研究(英文).pdf

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1、·760·ChineseMedicalJournal2001;114(7):760-763EpitopemappingonWegener'sgranulomatosisautoantigenproteinase3HUANGZhen黄真andChristopherMartinLockwoodKeywords:vasculitis·Wegener'sgranulomatosis·proteinase3·anti-neutrophilcytoplasmicautoantibodies·epitopeObjectiveTomapt

2、heepitopesonWegener'sgranulomatosisautoantigenproteinase3.MethodsAntigenicityofproteinase3wasstudiedwithWesternblotanalysisinwhichproteinase3waspreparedunderreducingandnon-reducingconditions.Twoanti-proteinase3monoclonalantibodies,HZ1F12andHZ1H3,wereusedtoinhibite

3、achotherandtoinhibit22anti-proteinase3positiveserafrompatientswithWegener'sgranulomatosisincompetitiveinhibitionenzyme-linkedimmunosorbentassays(ELISA)andWesternblotanalysis.ResultsAllmonoclonalantibodiesandpatientserarecognizedproteinase3undernon-reducingconditio

4、nsinWesternblotanalysis.HZ1F12wasinhibited74%byHZ1H3.10/22(46%)serawerecompletelyorpartiallyinhibitedbyHZ1F12;9/22(41%)serawerepartiallyinhibitedbyHZ1H3;and6/22(27%)wereinhibitedbybothmonoclonalantibodies.IninhibitionWesternblotanalysis,thebindingofpatientseratopr

5、oteinase3couldbeinhibitedbyHZ1F12.ConclusionsTheepitopesofWegener'sgranulomatosisautoantigenwereconformational.Anti-proteinase3monoclonalHZ1F12andHZ1H3recognizedsimilaroroverlappingepitopesontheproteinase3molecule.Epitopesofproteinase3recognizedbyanti-proteinase3p

6、ositiveserawerenotrestricted.ChinMedJ2001;114(7):760-763Wegener'sgranulomatosis(WG)isonetypeofsystemicMabsweretestedbyimmunofluorescencestaininginhumangranulocytesandgaveastainingpatternidenticaltothesmallvesselvasculitides,characterizedbynecrotizing2patternobtain

7、edwithserafrompatientswithWG.AffinityganulomatousinflammationoftheupperandlowerrespiratorychromatographypurifiedHZ1F12waslabeledwithalkalinetractwithpauci-immunenecrotizingcrescentphosphataseusingtheglutaraldehydemethod.Acommercialglomerulonephritisandvasculitisof

8、predominantlysmallmouseanti-myeloperoxidase(anti-MPO)monoclonalbloodvessels.Anti-neutrophilcytoplasmicantibodiesantibody(Dakopatts)wasusedasmousemonoclo