Serum and Antibodies of Glaucoma Patients Lead to Changes in the Proteome, Especially Cell Regulatory Proteins, in Retinal Cells 英文参考文献.docVIP

SerumandAntibodiesofGlaucomaPatientsLeadto ChangesintheProteome,EspeciallyCellRegulatory Proteins,inRetinalCells KatharinaBell,SebastianFunke,NorbertPfeiffer,FranzH.Grus* ExperimentalOphthalmology,DepartmentofOphthalmology,MedicalCenteroftheJohannesGutenbergUniversity,Mainz,Germany Abstract Purpose: Previous studies show significantly specifically changed autoantibody reactions against retinal antigens in the serumofglaucomaandocularhypertension(OHT)patientsincomparisontohealthypeople.Aspathogenesisofglaucoma still is unknown the aim of this study was to analyze if the serum and antibodies of glaucoma patients interact with neuroretinalcells. Methods:R28cellswereincubatedwithserumofpatientssufferingfromprimaryopenangleglaucoma(POAG),normal tension glaucoma (NTG) or OHT, POAG serum after antibody removal and serum from healthy people for 48h under a normal or an elevated pressure of 15000Pa (112mmHg). RGC5 cells were additionally incubated with POAG antibodies under a normal pressure. Protein profiles of the R28 cells were measured with Seldi-Tof-MS, protein identification was performedwithMaldi-TofTof-MS.ProteinanalysisoftheRGC5cellswasperformedwithESI-OrbitrapMS.Statisticalanalysis includingmultivariatestatistics,variancecomponentanalysisaswellascalculatingMahalanobisdistanceswasperformed. Results: Highly significant changes of the complex protein profiles after incubation with glaucoma and OHT serum in comparisontohealthyserumweredetected,showingspecificchangesinthecells(e.g.Proteinat9192Da(p,0.001)).The variancecomponentanalysisshowedaneffectoftheserumof59%onthecells.Thepressurehadaneffectof11%onthe cells.Antibodyremovalledtosignificantlychangedcellreactions(p,0.03).Furthermore,theincubationwithPOAGserum anditsantibodiesledtopro-apoptoticchangesofproteinsinthecells. Conclusions: These studies show that the serum and the antibodies of glaucoma patients significantly change protein expressionsinvolvedincellregulatoryprocessesinneuroretinalcells.Thesecouldleadtoahig