Scabies Mite Peritrophins Are Potential Targets of Human Host Innate Immunity 英文参考文献.docVIP

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Scabies Mite Peritrophins Are Potential Targets of Human Host Innate Immunity 英文参考文献.doc

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ScabiesMitePeritrophinsArePotentialTargetsof HumanHostInnateImmunity AngelaMika1.,PriscillaGoh1.,DeborahC.Holt2,DaveJ.Kemp1,KatjaFischer1* 1QueenslandInstituteofMedicalResearchandAustralianCentreforInternationalandTropicalHealthandNutrition,UniversityofQueensland,Brisbane,Queensland, Australia,2MenziesSchoolofHealthResearch,CharlesDarwinUniversity,Darwin,NorthernTerritory,Australia Abstract Background:PruriticscabieslesionscausedbySarcoptesscabieiburrowinginthestratumcorneumofhumanskinfacilitate opportunisticbacterialinfections.Emergingresistancetocurrenttherapeuticsemphasizestheneedtoidentifynoveltargets forprotectiveintervention.Wehavecharacterizedseveralproteinfamilieslocatedinthemitegutascrucialfactorsforhost- parasite interactions. Among these multiple proteins inhibit human complement, presumably to avoid complement- mediateddamageofgutepithelialcells.Peritrophinsaremajorcomponentsoftheperitrophicmatrixoftenfoundinthegut of arthropods. We hypothesized that a peritrophin, if abundant in the scabies mite gut, could be an activator of complement. Methodology/PrincipalFindings:Anovelfulllengthscabiesmiteperitrophin(SsPTP1)wasidentifiedinacDNAlibraryfrom scabiesmites.Theaminoacidsequencerevealedfourputativechitinbindingdomains(CBD).Recombinantexpressionof one CBD of the highly repetitive SsPTP1 sequence as TSP-hexaHis-fusion protein resulted in soluble protein, which demonstratedchitinbindingactivityinaffinitychromatographyassays.AntibodiesagainstarecombinantSsPTP1fragment wereusedtoimmunohistochemicallylocalizenativeSsPTP1inthemitegutandinfecalpelletswithintheupperepidermis, co-localizingwithserumcomponentssuchashostIgGandcomplement.EnzymaticdeglycosylationconfirmedstrongN- andO-glycosylationofthenativeperitrophin.Serumincubationfollowedbyimmunoblottingwithamonoclonalantibody against mannan binding lectin (MBL), the recognition molecule of the lectin pathway of human complement activation, indicatedthatMBLmayspecificallybindtoglycosylatedSsPTP1. Conclusions/Sign