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1、IMMUNOLOGICALINVESTIGATIONS,23(4&5),313-321(1994)HUMANCORONAVIRUSOC43INTERACTSWITHMAJORHISTOCOMPATIBILITYCOMPLEXCLASSIMOLECULESATTHECELLSURFACETOESTABLISHINFECTIONArleneR.CollinsDepartmentofMicrobiologySchoolofMedicine,StateUniversityofNewYorkatBuffaloABSTRACTHumancoronaviruseshavebeenassociate
2、dwithcommoncolds,diarrheaandenterocolitis,andhavebeenimplicatedinmultiplesclerosis.HLAclassImoleculesmayplayacriticalroleasreceptorforOC43becausemonoclonalantibody(mAb)W6/32toHLA-A,-Band-Cspecificitiescompletelyblocksinfectivityinhumanrhabdomyosarcoma(RD)cells.TheroleofHLAclass1antigenastheviru
3、sreceptorwasexaminedusingHLA-A3.1stablytransfectedhumanplasmacellsanduntransfectedHMY.ClRcellswhichdonotexpressHLA-Aand-Bmolecules.Whenthecells(5~10~)wereinfectedatamultiplicityofone,theHLA.A3transfectedcellsproduced108PFUofviruswhereasnoreplicationoccurredintheHMY.ClRcellsmAbW6/32reducedthevir
4、usyieldby99.9%CellForpersonaluseonly.membranesfromHMY.ClR,HMY.A3cellsandchickenerythrocyteswerebiotinylatedaslivecells.ImmunoprecipitationwithpolyclonalantiviralantibodytodetectbindingofbiotinylatedcellmembranestovirusrevealedthatbiotinylatedHMY.A3membranesco-precipitatedwithvirus-antibodycompl
5、exeswhentheimmunoprecipitateswereelectrophoresedonSDS-PAGEgel,electroblottedandstainedwithAvidin-horseradishperoxidase.TheresultsprovidedirectevidencethatOC43viruscanrecognizeHLAclassIasreceptoronthecellsurface.Keywords:HLAClassIreceptor-OC43virusInteractionImmunolInvestDownloadedfrominformahea
6、lthcare.combyNyuMedicalCenteron01/06/15Abbreviationsused:RD,humanrhabdomyosarcomacells;HLA,humanleucocyteantigen;HMY.A3.1,humanplasmacytomacellstransfectedwithHLA-A3gene,HMY.C1RhumanplasmacytomacellsthatlackHLA-AandBmoleculesonthecellsurface;MHC-I,majorhistocompatibilitycomplexclassI;Fbs,fetalb
7、ovineserum;pfu,virusplaqueformingunits;SephProA,proteinAsepharoseCL-4B;2-me,2-mercaptoethanol;mAb,monoclonalantibody;Avidin-HRP,avidinconjugatedtohorseradishperoxidase;4CN,4-chloro-1naphthol;CEA,carcinoembryonicantigen.313Copyrigh