ThisarticleispartoftheTopicalCollectiononImmunologicDiagnosticTestsinAllergyGabrieleGadermaierGabrieleGadermaiersbgacatIsabelPablosIsabelPablossbgacatSabrinaWildnerSabrinaWildnersbgacatCl ID: 939501
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IMMUNOLOGIC/DIAGNOSTICTESTSINALLERGY(MCHAPMANANDAPOMÉS,SECTIONEDITORS)PollenAllergensforMolecularDiagnosisIsabelPablosSabrinaWildnerClaudiaAsamMichaelWallnerGabrieleGadermaier1,2Publishedonline:22March2016TheAuthor(s)2016.ThisarticleispublishedwithopenaccessatSpringerlink.comAbstractPollenallergensareoneofthemaincausesoftypeIallergiesaffectingupto30%ofthepopulationinindustrial-izedcountries.Climaticchangesaffectthedurationandinten-sityofpollenseasonsandmaytogetherwithpollutioncon-tributetoincreasedincidencesofrespiratoryallergyandasth-ma.Allergenicgrasses,trees,andweedsoftenpresentsimilarhabitatsandfloweringperiodscompromisingclinicalanam-nesis.Molecule-basedapproachesenabledistinctionbetweengenuinesensitizationandclinicallymostlyirrelevantIgEcross-reactivitydueto,e.g.,panallergensorcarbohydratedeterminants.Inaddition,sensitivityaswellasspecificitycanbeimprovedandleadtoidentificationoftheprimarysensitizingsourcewhichisparticularlybeneficialregardingpolysensitizedpatients.Thisreviewgivesanoverviewonrelevantpollenallergensandtheirusefulnessindailypractice.Appropriateallergydiagnosisisdirectlyinfluencingdecisionsfortherapeuticinterventions,andthus,reliablebiomarkersarepivotalwhenconsideringallergenimmunotherapyinthecon-textofprecisionmedicine.KeywordsPollenallergensMolecule-baseddiagnosisGrasspollenallergensTreepollenallergensWeedpollenallergensIntroductionAllergicreactionstopollenrepresentthemostfrequenttypeIallergiesaffectingupto30%oftheindustrializedpopulationwww.eaaci.org).Climaticchangesareexpectedtoinfluencethedurationaswellastheintensityofpollenseasonswhichmightinhandwithairpollutioncontributetoincreasednumbersofrespiratoryallergyandasthma[].Inaddition,pollen-derivednanovesiclesandothersmallcomponents(e.g.,adenosine)mayplayaspecificroleinthecourseofallergicdiseases[].Clinicalanamnesisofpollenallergiestoidentifythedisease-elicitingsourcecanbehamperedbysimilarhabitatandfloweringperiodsofcertainplantsandthefactthatsymptomsmaybeelicitedbypollentransportedfarbythewind[].Inaddition,patientsareoftenmulti-sensitizedtodiverseallergensourcesduetoIgEcross-reactivity,co-sensi-tization,orboth[].SkinpricktestsandspecificIgEde-tectionusingcrudepollenextractsarecurrentlyperformedinroutineallergydiagnosis.Allergenextractshowevercontainavarietyofallergenicandnon-allergeniccomponents,andstan-dardizationofpollenextractsisdifficultduetovaryingsourcematerialandproductpreparations[].Besidessourcespecif-icandthusgenuinemarkerallergens,alsominorallergensincludingpanallergensaswellsasallergenswithcross- ThisarticleispartoftheTopicalCollectiononImmunologic/DiagnosticTestsinAllergyGabrieleGadermaierGabriele.Gadermaier@sbg.ac.atIsabelPablosIsabel.Pablos@sbg.ac.atSabrinaWildnerSabrina.Wildner@sbg.ac.atClaudiaAsamClaudia.Asam@sbg.ac.atMichaelWallnerMichael.Wallner@sbg.ac.atDepartmentofMolecularBiology,UniversityofSalzburg,HellbrunnerStraße34,5020Salzburg,AustriaChristianDopplerLaboratoryforBiosimilarCharacterization,UniversityofSalzburg,HellbrunnerStraße34,5020Salzburg,AustriaCurrAllergyAsthmaRep(2016)16:31DOI10.1007/s11882-016-0603-z reactivecarbohydrates(CCDs)arepresentindiagnosticex-tractswhichmightimpedeexactdiagnosis.Moleculardiag-nosisusingwell-characterizedpurifiedallergencomponentsfromnaturalsourceorproducedasrecombinantmoleculeallowsclinicianstoobtaindetailedinformationonsensitiza-tionprofilesandthussupportsimprovedpatientsmanage-ment[Morethan150pollenallergensareofficiallyacknowl-edgedbytheIUISallergennomenclaturesub-committeeorig-inatingfromgrasses,trees,andweeds(www.allergen.orgThisreviewfocusesonthemostrelevantandcommerciallyavailablecomponentswhicharediscussedinmoredetail(Tables).WealsoprovideanoverviewondiverseIgEcross-reactivityprofileswithintheOlee1-like,pectatelyaseandnon-specificlipidtransferprotein(nsLTP)families(Fig.MethodsforMolecule-BasedAllergyDiagnosisApartfromclinicalhistory,skinpricktests(SPT)usingaller-genextractsrepresentoneofthemostcommonusedinvi
vodiagnostictoolstoconfirmanimmediateIgE-mediatedaller-gicreaction.Sincethe1990s,theperformanceofseveralpu-rifiedrecombinantpollenallergenswasinvestigatedinprov-ocationtestsandcomparedtonaturalallergensandextractsts13].Thoughrefinementofdiagnosiswithpurifiedmole-culeswasproveneffective,invivodiagnosisusingcompo-nentsisnowadayswidelyrestrictedtoGMP-producedmate-rial.Alternativessuchasthebasophilactivationtest(BAT),aninvitromethodtomonitorupregulationofactivationmarkersCD63andCD203uponallergen-triggeredactivationofpri-marybasophils,ormediatorreleaseassayswithpassivelysen-sitizedbasophils,whichmeasureeitherhistamineor-hexos-aminidase,mayhelptocircumventthisproblem.Whiletheseassaysdemonstratefunctionalallergenresponses,theyareratherdemandingregardingthecostsandexperimentalskillsandthusarenotusedindailypractice.Measurementofspe-cificIgE(sIgE)inserumiscurrentlythemostwidelyusedinvitromethodforallergydiagnosis.Besidesallergenextractsprovidedforsingleplexanalyses(ImmunoCAP),apanelofpurifiednaturalandrecombinantcomponentsiscommerciallyavailableforroutinediagnosis.Inaddition,allergenmi-croarrayswithmorethan100purifiedallergenmole-cules(ImmunoCAPISAC)enablesimultaneousIgEmeasurementusingonlyminuteamountsofblood.Thesemultiplexassaysofferanattractivealternativetorefineallergydiagnosis,monitordiseaseprogressionaswellastherapeuticoutcomes[].AllinvitromethodsprovidereliableinformationonIgEsensitizationwhilecorrelationwithclinicallyrelevantsymptomsremainschallenging[ Table1GrasspollenallergensFloweringperiodGrassgroup1(Beta-expansin)Grassgroup2(expansin-likeprotein)Grassgroup3(expansin-likeprotein)Grassgroup4(Berberine-bridgeenzyme)Grassgroup5(Ribonuclease)Grassgroup6PolcalcinOlee1-likeproteinProfilinGrassgroup13(Polygalacturonase)Timothygrass5Phlp1Phlp2Phlp3Phlp4Phlp5Phlp6Phlp7Phlp11Phlp12Phlp13PhleumpratensePerennialryegrass5Lolp1Lolp2Lolp3Lolp4Lolp5Lolp11LoliumperenneOrchardgrass5Dacg1Dacg2Dacg3Dacg4Dacg5DactylisglomerataKentuckybluegrass5Poap1Poap5PoapratensisBermudagrassallseasonCynd1Cynd7Cynd12CynodondactylonBahiagrassallseasonPasn1PaspalumnotatumJohnsongrassallseasonSorh1Sorh2Sorh13SorghumhalepenseMajorallergenshighlightedinitalicsAvailableforsingle-andmultiplexanalysisAllergennotofficiallyacknowledgedbytheWHO/IUISallergennomenclaturesub-committee Page2of12CurrAllergyAsthmaRep(2016)16:31 Table2RelevanttreeandweedpollenallergensFloweringperiodBetv1-likeprotein(PR-10)ProfilinPolcalcinPhenylcoumaranbenzylicetherreductase-likePolygalacturonasePlantinvertase/pectinmethylesteraseinhibitor1,3beta-glucanase(PR-2)Defensin-prolinfusion(PR-12)nsLTP(PR-14)Olee1-likemethylesteraseCysteineproteaseTREEPOLLENBirch3Betv1Betv2Betv3Betv6BetulaverrucosaBetv4AlderAlng1Alng4AlnusglutinosaHornbeam4Carb1CarpinusbetulusHop-hornbeam4Ostc1OstryacarpinifoliaHazelnut2Cora1Cora2Cora6CorylusavellanaFags1FagussylvaticaChestnutCass1CastaneasativaQuea1QuercusalbaLondonplanetree4Plaa2Plaa1Plaa3PlatanusacerifoliaOlive46Olee2Olee3Olee12Olee9Olee7Olee1Olee11OleaeuropeaOlee8Olee10EuropeanashFrae1FraxinusexcelsiorCommonprivetLigv1LigustrumvulgareSyrv1SyringavulgareMediterraneancypress1Cups1CupressussempervirensArizonacypress8Cupa1CupressusarizonicaJapanesecypress1Chao3Chao1ChameocyparisobtuseJapanesecedar2Cryj2Cryj1CryptomeriajaponicaMountaincedar12Juna2Juna1JuniperusasheiWEEDPOLLENRagweed79Amba8Amba9Amba1Amba4Amba6Amba11AmbrosiaartemisiifoliaAmba10MugwortArtv6Artv1Artv3ArtemisiavulgarisFeverfew7-9Parh1PartheniumhysterophorusPellitoryallseasonParj3Parj4Parj1ParietariajudaicaParj2Englishplantain4Plal1Plantagolanceolata CurrAllergyAsthmaRep(2016)16:31Page3of12 GrassPollenAllergensGrassesarecultivatedforfood,animalfodder,surfacevege-tation,andmeadows,butalsorepresentoneofthemajorcausesofrespiratoryallergiesworldwide[].Theclini-callymostrelevantsourceofpollenallergensisfoundwithinthePoaceaefamily.Amongthis,thePooideaesubfamilyisprevalentintemperateclimatesandthemostabundantaller-genicspecieswithintheP
ooideaeareTimothygrass,Perennialryegrass,Orchardgrass,andKentuckybluegrass.Thepollinationseasontypicallystartsinspringandlastsuntillatesummer,pollenpeaksareusuallyfoundbetweenJuneandJuly.Insubtropicalregions,membersoftheChloridoideaeandPanicoideaesubfamiliesarewidelydistributed.Here,themostimportantspeciestriggeringpollenallergyareBermudagrass,Bahiagrass,andJohnsongrass[ Table2FloweringperiodBetv1-likeprotein(PR-10)ProfilinPolcalcinPhenylcoumaranbenzylicetherreductase-likePolygalacturonasePlantinvertase/pectinmethylesteraseinhibitor1,3beta-glucanase(PR-2)Defensin-prolinfusion(PR-12)nsLTP(PR-14)Olee1-likemethylesteraseCysteineproteaseGoosefoot610Chea2Chea3Chea1ChenopodiumalbumRussianthistle79Salk4Salk5Salk1SalsolakaliAmaranth79Amar2AmaranthusretroflexusAnnualmercury5Mera1MercurialisannuaMajorallergenshighlightedinitalicsCommerciallyavailableforsingle-andmultiplexanalysisCommerciallyavailableformultiplexanalysisonlyCommerciallyavailableforsingleplexonlyAllergennotofficiallyacknowledgedbytheWHO/IUISallergennomenclaturesub-committeeSequencehomologytoN-terminusofOlee9,listedascarbohydrate-bindingmolecule a Ole e 1-like familyb Pectate lyasesc Lipid transfer proteinsOle e 1 Fra e 1 87 Pla l 1 37 38 Che a 1 37 38 34 Sal k 5 37 39 36 72 Phl p 11 31 34 29 46 45 Ole e 1 Fra e 1 Pla l 1 Che a 1 Sal k 5 Phl p 11 Amb a 1 Art v 6 59 Cup a 1 46 48 Cup s 1 44 46 95 Cry j 1 46 47 79 78 Jun a 1 45 46 95 96 79 Amb a 1 Art v 6 Cup a 1 Cup s 1 Cry j 1 Jun a 1 Art v 3 Amb a 6 35 Par j 2 30 33 Ole e 7 25 29 10 Pla a 3 53 37 32 33 Pru p 3 49 36 24 20 53 Art v 3 Amb a 6 Par j 2 Ole e 7 Pla a 3 Pru p 3 Fig.1Sequenceidentitymatrixand3D-modelsofallergenicproteinfamilies.Olee1-likeproteinsandstructureofPlal1.0101(4Z8W),pectatelyasesandmodelofAmba1.0101(template1PXZ),andlipidtransferproteinsandmodelofArtv3.0201(template2B5S).MultiplesequencealignmentsperformedinClustalOmega.ModelsweregeneratedusingSwiss-Model(www.swissmodel.expasy.org),andribboncartonsareshownusingUCSFChimera(www.cgl.ucsf.edu/chimeraGreenboxesrepresentdemonstratedIgEcross-reactivity,lightgreenboxesrepresentpotentialIgEcross-reactivitybasedonhighsequenceidentity,redboxesrepresentno/limiteddemonstratedIgEcross-reactivityandlightredboxesrepresentno/limitedIgEcross-reactivitybasedonlowsequenceidentity Page4of12CurrAllergyAsthmaRep(2016)16:31 Pollinationofthesegrassespersiststhroughouttheyearpeakingfromearlysummerthroughautumn.Climatologicalchangesduetoglobalwarmingandhu-manactivitieshaveadditionallyinfluencedpollendistri-butionaswellaspotencywhichfurtherincreasessensi-tizationrisks[TimothyGrassTimothygrassiswidelydistributedinthetemperateclimateandoneofthebestcharacterizedallergenicgrasses.ThehighestsensitizationprevalenceisregisteredintheEuropeanpopulation,rangingfrom18.5to28.5%[].Aprevalenceof36%amongAustrianpupilswasrecentlyfoundusingaller-genmicroarrayanalysis(Stemesederetal.,unpublished).Themajorityofcommerciallyavailablecomponentsforgrasspol-lenallergyoriginatefromTimothygrass(Table).Inthiscontext,Phlp1,amajorallergenfromTimothygrasswithasensitizationprevalenceof�90%amonggrasspollenallergicpatients,seemsextremelyimportant[].Phlp1-specificIgEisconsideredamarkerforgenuinesensitizationandwasrecentlyalsoproposedasinitiatormoleculeforTimothygrasspollenallergy[].InthestudybyHatzleretal.,75%ofgrasspollenallergicpatientsstartedwithmono-sensitizationtoPhlp1,insomecasesyearsbeforeonsetofclinicalsymptoms,whichsubsequentlydevelopedintopoly-sensitizationtowardsothergrasspollenaller-gens[ThesecondmajorallergeninthissourceisPhlp5,trigger-ingspecificIgEin6590%ofgrasspollenallergicpatients.TogetherwithPhlp1,itisconsideredamarkermoleculeforgrasspollenallergy,andinrarecasesofabsentPhlp1-specificIgE,Phlp5isgivingreliableresultsofgenuinesensitizationn21,22].Phlp5ishighlycross-reactivewithothergroup5grasspollenallergens,whicharerestrictedtograssesofthePooideaesubfamily[].AhighsequencehomologywithPhlp6isobse
rved,andsignificantlevelsofIgEcross-reactivityhavebeendemonstrated[].Phlp6isfrequentlyusedtogetherwithPhlp2andPhlp5toconfirmspecificsensitizationtoPooideaegrasses[].However,careshouldbetakenwithinterpretationofPhlp2resultssinceahomol-ogousmoleculetermedSorh2wasrecentlyidentifiedinJohnsongrass,amemberofthePanicoideaesubfamilysubfamily25].ThetentativelytermedTimothygrasspollenallergenPhlp3showssequenceidentitywithPhlp2andpartiallywiththeC-terminusofPhlp1;however,whilecross-reactivitybetweengroup2and3grasspollenallergenswasdemonstrated,relevantcross-reactivitytogroup1allergensisnotevident[Amonggrasspollencomponents,solelyPhlp4isprovidedasnaturalproteinharboringCCDs.RecombinantPhlp4dem-onstrateslowerIgEreactivity,thusIgE-bindingtocommer-ciallyavailablenaturalPhlp4shouldbeanalyzedforCCDsreactivity[].InTimothygrass,thepanallergensPhlp7(polcalcin)andPhlp12(profilin)arepresentprobablyinducingIgEcross-reactivitywithhomologsfromtrees,weedsandfoods[].IntheabsenceofPhlp1-and/orPhlp5-specificIgE,sensitizationtopanallergensisnotindicativeofgenuinegrasspollenreactivity[less,anassociationofthePhlp12sensitiza-tionandthedevelopmentoforalallergysyndromesweredemonstratedinItalianchildren[].Duetothelowsequencesimilarity,IgEcross-reactivityofPhlp11withotherOlee1-likefamilymembersfromtreesandweedsisabsentorverylimited[](Stemesederetal.,unpublished).PerennialRyegrass,OrchardGrass,andKentuckyBlueGrassAllergensfromPerennialryegrass,Orchardgrass,andKentuckybluegrasshavebeenidentifiedandarelistedinTable,butnoneofthoseiscommerciallyavailablefordiag-nosis.DuetohighsequenceidentityandextensiveIgEcross-reactivityofallergenswithinthePooideaesubfamily,diagno-sistotemperategrassesisusuallyperformedwithcomponentsfromTimothygrass[BermudaGrassCynd1,themajorallergenfromBermudagrass,iscurrentlytheonlycommerciallyavailablemoleculeofthesubtropicalgrasses.ItisconsideredasbiomarkerforgenuinesensitizationtotheChloridoideaesubfamily[].Cynd1isrecognizedby76100%ofgrasspollenallergicpatients,whichmayalsoresultinIgEcross-reactivitywithPhlp1[].Therefore,itwassuggestedthattheprimarysensitizationtoBermudagrassisindicatedwhenspecificIgE-levelstoCynd1exceedIgE-bindingtoPhlp1andIgEtoPhlp5isnotdetectable.SinceCynd1isharboringCCDs,glycan-derivedreactivityshouldbemonitoredduringdiagnosis[BahiaGrassandJohnsonGrassCurrently,noallergenfromthePanicoideaesubfamilyiscom-merciallyavailableformoleculardiagnosis.Inrecentyears,agrowingnumberofallergensfromthesesourceshavebeendescribed.Timbrelletal.showedimprovedsensitivityandspecificitywhenusingnPasn1formoleculardiagnosisofBahiagrasspollenallergicpatients[].Theauthorssug-gestedtoincludethePasn1forcommerciallyavailablediag-nosisofBahiagrassallergyasasub-groupofpatientsshowedspecies-specificIgEandTcellreactivity[].Asmen-tionedabove,Sorh2wasrecentlyidentifiedinJohnsongrasswhichrefutestheassumptionthatgroup2allergensareabsentinsubtropicalgrasses[ CurrAllergyAsthmaRep(2016)16:31Page5of12 TreePollenAllergensIngeneral,treesarewoodyperennialplantswithanelongatedself-supportingstemortrunkandsupportingbranchesthatformamoreorlessdefinedcrown.Todistinguishtreesfromshrubs,acertainstemheightordiameterissometimesusedasdecisioncriteria.Treesdonotcompriseasingletaxonomicgroupandincludevariousspeciesthatseparatelyevolvedstemsandbranches[].Mostoftheclinicallyrelevantpollenallergensareproducedbywind-pollinatedtreesbelongingtoonlyfourdifferentorders,whichshowanalmostworldwidedistribution.Fordetailsontreepollenallergens,reviewAsametal.[BirchandRelatedFagalesSpeciesClassifiedwithintheorderofFagales,birchandtherelatedtreespeciesalder,hornbeam,hop-hornbeam,hazelnut,beech,chestnut,andoakconstitutethemaincauseofearlyseasonalrhinitisinthetemperateclimatezoneoftheNorthernHemisphere[].Worldwide,morethan100millionpatientssufferfrombirchpollenallergiesandinEuropeclinicallyrelevantsensitizationtobirchaffectsaround19.6%oftheallergicpopulation.Clinicallyrelevantsensitizationratestohazelandalderpollen
werereportedat17.1and16.2%,re-spectively[].ThemajorallergensfromFagalestreesareclassifiedasmembersofthepathogenesis-related-10(PR-10)proteins,andBetv1frombirchpollenisgenerallyac-knowledgedasmainsensitizerandmarkerallergenofthisfamily(Table).Amongbirchpollenallergicindividuals,sensitizationratestoBetv1ofupto90%ormorehavebeenreported[].However,inhibitionexperimentsrevealedthatbesidesbirch,severalotherFagalesspeciesmighthavethepotentialtosensitizesusceptibleindividuals[].Thisseemsespeciallyimportantinareaswherebirchtreesarevir-tuallyabsent,thusexposuretootherallergenicFagalespollenwilleventuallyleadtosensitizationandthedevelopmentofFagalespollenallergy.Currently,Betv1istheonlygroup1Fagalesallergenofferedforsingleplex,whileAlng1andCora1areadditionallyavailableformultiplexanalysis.Sinceoakpopulationsareverywide-spreadnotonlyinEuropebutalsoinvastpartsofNorthAmericaandAsiaanddonotnecessarilycoexistwithbirchorhazel,Quea1shouldbeconsideredfordiagnosisespeciallyinthoseareas.Morethan70%ofbirchpollenallergicpatientsdevelopadversesymptomstofoodsuchasfruits,nuts,orvegetables[].Thisclinicalconditionoftenreferredtoaspollen-foodsyndromeal-waysinvolvesapre-sensitizationtoFagalespollenal-lergens[].Therefore,diagnosisofBetv1-relatedfoodallergiesisdirectlylinkedtoacorrectdiagnosisoftheunderlyingpollenallergy.Moreover,severalmi-norallergens,amongthemthepanallergensprofilinandpolcalcin,havebeenidentifiedinmanyallergenicFagalesspecies,evokingsensitizationratesof44.6and9.4%,respectively[PlaneTreePlanetreesbelongtotheorderofProtealesandarenativetotheNorthernHemisphere.Theypreferentiallygrowintem-perateregionsfromAsiatoEuropeandNorthAmerica.Duetotheirresistancetowardspollution,theyarewidelyusedasornamentaltreesinurbanareasandatroadsides[].With3.3%intheallergicpopulation,clinicallyrelevantsensitiza-tionthroughoutEuropeisratherlowbutcanreachalmost12%intheUK[].Morethan80%ofplane-sensitizedpatientsreacttothemajorallergensPlaa1(plantinvertaseinhibitor)andPlaa2(polygalacturonase)[].TheminorallergenPlaa3isamemberofthensLTPfamilyandshowshighcross-reactivitywithPrup3,themajorallergenfrompeach[Olive,Ash,Lilac,andPrivetThemajorallergy-elicitingspecieswithintheorderofLamialesareolive,ash,lilac,andprivet.Olivetreesarepref-erentiallycultivatedintheMediterraneanareawhereupto70%ofpatientswithrespiratorysymptomsaresensitizedtoolivepollen[].Withasensitizationprevalenceof80%amongolivepollenallergicpatients,Olee1representstheallergenandthusistheeponymoftheOlee1-likeproteinfamilywithhomologsintree,grass,andweedpollen.Whilecross-reactivitytohomologuesinotherOleaceaetreesisveryhigh,cross-reactivitywithgrass(Phlp11)andweed(e.g.,Plal1)Olee1-likeallergensislimited[(Stemesederetal.,unpublished).BesidesOlee1,10otherolivepollenallergens,includingthepanallergensOlee2(profilin)andOlee3(polcalcin),havebeendescribedanddetailscanbereviewedinVillalbaetal.[].Dependingongeographiclocationandthusontheexposurelevelofthepopulation,theallergensOlee6,7,10,and11mayeventuallybecomemajorallergensofolivepollen[IncentralEurope,ashpollenmaycausesensitizationratesfrom4to18%[].ThemajorallergenofEuropeanash,Frae1,hasbeenclassifiedasmemberoftheOlee1-likefamilyandshowshighsequencesimilaritytootherOlee1-likeproteinsfromLamiales[].ArecentstudyfromImhofetal.confirmedthehighcross-reactivitybetweenOlee1andFrae1andsuggestedusingOlee1asdiagnosticmakerofashpollenallergy[].Moreover,a-1,3-glucanasefromashtermedFrae9wasrecentlyidentified,whichcanaccountforasensitizationprevalenceupto60%indistinctareasofFrance[].CommonprivetaswellaslilacareoftenusedforornamentalpurposesandfrequentlyfoundinEurope,Asia,andNorthAmerica.Bothexpresshighlycross-reactivemajorallergensbelongingtotheOlee1-likeproteinfamily(Ligv1 Page6of12CurrAllergyAsthmaRep(2016)16:31 andSyrv1,respectively);sensitizationratesarehowevergenerallylow[AllergenicCypressSpecies,JapaneseCedar,andJuniperTreesTheorderofPinalescomprisesallergeniccypress,cedaraswellasjuniperspecies.Incontra
sttothepreviouslydescribedallergenictrees,PinalesaregymnospermswidelydistributedthroughouttheNorthernHemisphere.MediterraneancypressanditsAmericanrelative,Arizonacypress,whichisprimarilynativetothesouth-westofNorthAmerica,arecloselyrelatedtoevergreentreeswhichcoexistintheMediterraneanareawww.eol.org).AmongtheallergicindividualsinEurope,2.6%showaclinicallyrelevantsensitizationtocypress,whichisgenerallylow,however,mayreachlevelsofupto42.7%asreportedforallergicpatientsinItaly[].Themajorallergensofbothspecies,Cups1andCupa1,belongtothepectatelyasefamilyandshare95.1%sequenceidentity.Moreover,clinicallyrelevant,highlycross-reactive,allergenicpectatelyaseshavebeenidentifiedasmajorallergensinMountaincedar(Juna1)endemicintheUSA,aswellasinJapanesecypress(Choo1)andJapanesecedar(Cryj1).ThelattertwospeciesformdominantpopulationsontheJapaneseislandisland37].SensitizationratesevokedbyChao1orthemorepotentCryj1affectupto40%ofthepopulationincertainareasofJapan[],whereassensitizationtoJuna1representsaserioushealthriskinTexasandMexico[Moreover,thepolygalacturonasesJuna2,Cryj2andChao2,havebeenidentifiedasmajorallergensintheseCupressaceaespeciesbeingresponsibleforsensitizationratesofupto80%[].Nevertheless,allergenicpolygalacturonasesdidnotgetmuchattentionsofarbutshouldeventuallybeconsideredascandidatesforallergydiagnosis.WeedsPollenAllergensThetermweeddoesnotrefertoanyspecificbotanicalfam-ilybutratherdescribesplantsoutsidetheorderoftreesorgrasses.Theyareusedasculinaryherbs,medicinalplants,andarefrequentlyecologicallyadaptivesegetalplants[].Climaticchangesaregenerallyimpactingtheflora,whichmightbepar-ticularlyadvantageousforweedsastheyareabletodominategroundcover,adapttovariousenvironments,orresideinecologicniches.Weedpollenallergicpatientsarefrequentlypoly-sensitizedtodiverseplantsources,thusmolecule-basedap-proachesareespeciallyvaluableforprecisediagnosis.Acom-prehensiveoverviewonproductionandbotanicalclassificationofweedpollenallergensisprovidedinGadermaieretal.andVillalbaetal.[RagweedShortragweedisnativetoNorthernandCentralAmerica,whereitisoneofthemajorelicitorsoftypeIpollenallergy.Sensitizationprevalencecanbeupto15.3%inthegeneralpopulationandtheweeddemonstrateshighcross-reactivitywithotherAmbrosiaspp.[].Sinceitsintroductionasballastgrain,ragweedalsobecamearelevantallergensourceinpartsofEurope,Asia,andAustralia[].GlobalwarmingalreadysubstantiallyprolongedtheragweedpollenseasonandfurtherspreadingoftheweedtotheNorthispre-dictedbyinsilicomodels[ThepectatelyaseAmba1ishighlyabundantinragweedpollenandrepresentsthedominantallergen.Basedonasen-sitizationprevalenceof�95%,thenaturalallergeniscurrentlyconsideredthemarkerallergenforgenuineragweedsensitiza-tion[].Incontrasttolackingcross-reactivitytohomo-logsfromtheCupressaceaefamily,antibodycross-reactivitywithArtv6frommugwortpollenisobserved.However,inhibitionassaysandTcellstudiesprimarilypointatastron-gerallergenicpotentialandsensitizingroleofAmba1[].Inarecentstudy,primarysensitizingcapacityofArtv6wasalsodemonstratedandtheauthorsconcludedthattrueco-sensitizationofragweedandmugwortisgenerallyrare[Recently,acysteineproteasewithstructuralhomologytogroupIallergensfromhousedustmiteswasidentifiedinragweedanddesignatedAmba11[].Itrepresentsalsoamajorallergensince66%ofpatientsreactedtothemolecule,whichwaspreviouslyhiddenintheAmba1fraction.Thoughthemajorityofallergenicreactivityisclearlyattribut-edtoAmba1andisolatedAmba11reactivityseemsrare,itcouldberelevantforincreasingthediagnosticpanel[TheminorragweedallergenAmba4consistsofadefensin-likedomainfusedwithaproline-richregionwithhomologyandIgEcross-reactivitytoArtv1frommugwortpollen[Ragweedallergensbelongingtotheprofilin,polcalcin,andnsLTPfamilies(Table)areaccountingformoderatetolowIgEreactivity[].Recenttranscriptomeandimmunoproteomedatasuggest68and41%IgEreactivityforragweedpolygalacturonaseandenolase,respectivelyly71],whichtogetherwithAmba3mightbeworthinvestigat-ingatthemoleculele
vel.MugwortCommonmugwortisendemicintheNorthernHemisphereandAustraliaandrepresentsthebeststudiedplantwithinthegenus.Amongpollinosispatients,sensitizationtomugwortpollenrangesbetween10and14%intheEuropeanandAsianpopulation[].Habitatandpollinationofmugwortarevastlyoverlappingwithragweedandthusconfoundingaller-gyanamnesis.Artv1,consistingofadefensin-likedomainfusedwithaproline-richregion,representsthemajorallergen CurrAllergyAsthmaRep(2016)16:31Page7of12 withasensitizationprevalenceof7095%[].Themark-erallergendemonstratespartialIgEcross-reactivitywithAmba4fromragweedandParh1fromfeverfewpollen.Primarysensitizationseemstobepredominatelyevolvingbythemajormugwortallergen(Pablosetal.,unpublisheddata).Inaddition,Artv3isofparticularinterestinregionswherensLTP-relatedfoodandpollenallergiesareobserved[].WhileArtv3mightpavethewaytofood-relatedsymptomsinpeachallergicpatients[],recentdataalsosuggestthatitisabletobonafideelicitrespiratorysymptomsinpatients[].Inmugwortsensitizedpatients,Artv6inhandwithArtv1isconsideredhighlyindicativeforprimarymugwortsensitization[PellitoryPollenofpellitoryisresponsibleforallergicreactionspredom-inatelyinMediterraneanregionswithsensitizationfrequen-ciesreaching6090%[].Duetoclimaticchanges,floweringperiodsthroughouttheyearwereregistered[ThemainallergencomponentsareParj1andParj2,bothbelongingtothensLTPfamilyshowingasensitizationfre-quencyofupto95%[].Theyareconsideredmarkerallergensforgenuinepellitoryallergy,sincetheydonotcross-reactwithhomologsfromotherpollenandfood[].Parj2showedhighsensitivityandspecificityandiscurrentlyusedforroutinemolecule-baseddiagnosisofpellitory[EnglishPlantainPlantagospp.areworldwideabundantweedsshowingrecur-rentfloweringseasonsoverlappingwithgrasses.Studiesdem-onstratedahighclinicalrelevanceinpartsofsouthernandcentralEurope[].ThemajorallergenPlal1belongstotheOlee1-likefamilyandisrecognizedby�90%ofpatients[].SinceIgEcross-reactivitywithotherfamilymembersislimited,itrepresentsahighlyspecificmarkerformolecule-baseddiagnosis[](Stemesederetal.,unpublished).RussianThistleandGoosefootTheseweedsareconsideredinvasivespeciesfoundinaridregionsoftheNorthernHemisphereandAustralia[Duetotheirprevioususeingreeningprograms,theyarehigh-lyabundantinMiddleEasterncountrieswheretheycanrep-resentmajorsensitizersforrhinitisandasthma[AmaranthaceaepollenalsogainedallergologicalrelevanceinareasofSpain,andtheyareexpectedtoplayanincreasinglyimportantrole[].ThepectinmethylesteraseSalk1accountsforthemajorityofIgEreactivitytoRussianthistleandwasshowntobeamarkerforgenuinesensitizationasitlowsdiscriminationfromchenopodsensitization[NaturalglycosylatedSalk1iscurrentlyavailableformolecule-baseddiagnosis;however,theuseofarecombinantmoleculemightbesuperiorasCCDscanbeavoided[].Salk5,amemberoftheOlee1-likeproteinfamilyshowed30%sensitizationprevalenceandconsiderablehighIgEcross-reactivitywithChea1fromchenopod[].Notably,thisIgEreactivityseemsspecificforAmaranthaceaeandnotre-latedtoOlee1fromolivepollen[Chea1belongstotheOlee1-likefamilyandpresentsasensitizationfrequencyof�70%amongchenopodallergicpatients[].HighIgEreactivitytochenopodprofilinandpolcalcinisnoted,anddiagnosticspecificitywasin-creasedbyusingacocktailofthreepurifiedproteins[However,thehighIgEcross-reactivityofthepanallergensrathersuggeststhatChea1shouldbeprimarilyconsideredfordiagnosisalthoughdiscriminationfromRussianthistlesensitizationseemschallenging[AnnualMercuryAnnualmercurywasreportedtobeanimportantsourceofpollinosisindistinctregionsofMediterraneancountriescountries99].Fordiagnosticpurpose,theprofilinMera1iscurrentlyavailableinmultiplexassays;however,duetohighIgEcross-reactivitywithotherpollenprofilins,interpretationoftheseresultsmightbelimited.PollenPanallergensThetermpanallergenreferstotheubiquitousdistributionoftheseallergensindiversesourcesandprofilinsandpolcalcinsareprototypicexamples[100].Theclinicalrelevanceisconsid-eredlimited,butthebroadIgEreactivityhighlyinfluenc
esextract-baseddiagnosis[101].Apanelofprofilinsandpolcalcins,consideredrepresentativesoftheentirepanallergengroup,isavailableformolecule-baseddiagnosisallowingdis-criminationbetweengenuineandpanallergensensitization(Tablesand).Forinvivodiagnosis,naturalpanallergensfrompalmtreewerepreviouslyused[102].Incontrast,mem-bersofthepectatelyase,nsLTP,andOlee1-likefamiliesdem-onstrateheterogeneousreactivitypattern.IgEcross-reactivityismostlyconfinedtocloselyrelatedmoleculeswithhighse-quenceidentityandmightvaryindifferentpopulations(Fig.ConclusionsDiagnosisofpollenallergiesmainlyreliesoncarefulanamne-sis,whichincludesnarrowingdownthesensitizingplantbythetimeclinicalsymptomsareobserved.However,floweringperiodsarefrequentlyoverlappingandcanvaryindistinctgeographicregionswhileclimaticchangesareexpectedtofurtheraggravatetheproblem[].Pollenallergic Page8of12CurrAllergyAsthmaRep(2016)16:31 patientsaretypicallyreactingtomorethanonesource,thusspecificdiagnosisisappliedeitherusingallergenextractsorpurifiedcomponents.Basedonmolecule-basedapproaches,morespecificresultsareobtainedandclinicallyirrelevantsensitizationsdueto,e.g.,panallergensorotherIgEcross-reactivecompoundscanbecircumvented[].Thecurrentlyavailablepanelofallergencomponentsiscoveringmostofthecommonallergensources;however,incomparisontoextract-basedproducts,thereisasubstantialgapinquantity.Allergyresearchwillcertainlyfurtherincreasethepanelofrelevantallergensofknownsourcesaswellasnovelallergensfromplantsthatwerelessexploredfordiagnosis[].Molecule-baseddiagnosiscanimprovesensitivity,specificity,predictseverityofreactions,andidentifythegenuinesensitizingsource[103].Thisrefineddiagnosiswasrecentlyshowntochangealargeproportionofallergenimmunotherapypre-scriptionasopposedtorelyingonanamnesisandSPTalonee101].Infuture,allergydiagnosisusingreliablebiomarkersmightbecrucialwhenconsideringallergenimmunotherapyinthecontextofprecisionmedicine[104AcknowledgmentsOpenaccessfundingprovidedbyAustrianScienceFund(FWF).ThefinancialsupportbytheAustrianScienceFund(I1152,P27589,andL688),theAustrianFederalMinistryofScience,ResearchandEconomy,theNationalFoundationofResearch,TechnologyandDevelopment,LandSalzburg,andthepriorityprogramAllergy-Cancer-BioNanoResearchCentreoftheUniversityofSalzburgisacknowledged.CompliancewithEthicalStandardsConflictofInterestDr.GadermaierreportspersonalfeesfromThermoFisherScientific.Drs.Pablos,Wildner,Asam,andWallnerdeclarenoconflictsofinterest.HumanandAnimalRightsandInformedConsentThisarticledoesnotcontainanystudieswithhumanoranimalsubjectsperformedbyanyoftheauthors.OpenAccessThisarticleisdistributedunderthetermsoftheCreativeCommonsAttribution4.0InternationalLicense(http://creativecommons.org/licenses/by/4.0/),whichpermitsunrestricteduse,distribution,andreproductioninanymedium,providedyougiveappro-priatecredittotheoriginalauthor(s)andthesource,providealinktotheCreativeCommonslicense,andindicateifchangesweremade.ReferencesPapersofparticularinterest,publishedrecently,havebeenhighlightedas:OfimportanceOfmajorimportance1.D'AmatoGetal.Effectsonasthmaandrespiratoryallergyofclimatechangeandairpollution.MultidiscipRespirMed.2015;10:39.2.FukuokaAetal.Dieselexhaustparticlesexacerbateallergicrhi-nitisinmicebydisruptingthenasalepithelialbarrier.ClinExpAllergy.2015.3.WimmerMetal.Pollen-derivedadenosineisanecessarycofactorforragweedallergy.Allergy.2015;70(8):94454.4.PradoNetal.Pollensomesasnaturalvehiclesforpollenallergens.JImmunol.2015;195(2):4455.ScevkovaJetal.InfluenceofairbornepollencountsandlengthofpollenseasonofselectedallergenicplantsontheconcentrationofsIgEantibodiesonthepopulationofBratislava,Slovakia.AnnAgricEnvironMed.2015;22(3):4516.NewsonRBetal.Geographicalvariationintheprevalenceofsensitizationtocommonaeroallergensinadults:theGA(2)LENsurvey.Allergy.2014;69(5):64351.7.PanznerPetal.Acomprehensiveanalysisofmiddle-Europeanmolecularsensitizationprofilestopollenallergens.IntArchAllergyImmunol.2014;164(1):7482.8.
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