PDF - World Allergy Organization Journal

192 Pages·2012·2.31 MB·English

A
BSTRACTS
Abstracts of the XXII World Allergy Congress,
–
4 8 December, 2011 Cancu´n, Me´xico
ORAL ABSTRACT SESSION Methods:301(PP)patientswithconfirmedrhinitisand/orrhinoconjunctivitis
ALLERGEN IMMUNOTHERAPY 1 weretreatedinadouble-blindstudywith4dosesof100DPP/mL,1000DPP/
mL,5000DPP/mL and 10.000DPP/mLallergenextract over22 weeks in
Germany,Poland,andLithuania.A1-daybuild-upphaseapplying0.1mL
and2times0.2mLallergenextractwasfollowedbyamaintenanceperiod
1
applying 0.5 mL in 3 to 4 weeks intervals. Before treatment a CPT was
Ultra-RushSpecificImmuneTherapywithDepigmentedand
performedwithsemi-logarithmicallyincreasingdosesupto10,000SQ-U/mL
PolymerizedAllergenExtractsisEffectiveandSafeinPatients
ofnativebirchextract,aftertreatmenttheCPTwasrepeatedwithdosesupto
WithSevereSARHighlySensitizedagainstPollenAllergens
100,000SQ-U/mL.Themainparameterwasthepercentageofpatientswith
MichaelaGruber,andUlrichAmon,MD.PsoriSolClinicforDermatology anincreaseofallergenextracttoprovokeapositiveCPTafterSIT.Secondary
&Allergy,Hersbrcuk,Germany. parameters were specific IgE, IgG1 and IgG4 as well as safety. The main
Background:Proceduresforspecificimmunetherapy(SIT)varywidely.We parameterwasinvestigatedusingahierarchictestprocedurecomparingthe
wereinterestedwhetherapre-seasonalultra-rushtreatmentwithdepigmented highest dose against the lowest, if statistically significant testing the next
andpolymerizedallergenextractsiseffectiveandsafeinpatientswithsevere lowerdoseagainstthelowestuntilthedifferencewasnolongersignificant.
SAR. Results:AnincreaseinallergenamounttoprovokeapositiveCPTafterSIT
Methods:31pts(21f,10m,meanage39.7years,range:17–69years)with was reached in 37.5% of the 100 DPP/mL, 50.7% of the 1000 DPP/mL,
severeSARandpartialasthmaattacks(31.3%)duringpollenseason(mean 54.9%ofthe5000DPP/mLand55.8%ofthe10,000DPP/mLgroup.Results
totalIgElevel152kU/L,range:5–5113kU/L;meanPhadiaCAPclass3.7 comparedtothelowestdosewerestatisticallysignificantforthe5000DPP/
and3.5againstbirchpollenandBetv1,resp.;meanPhadiaCAPclass3.7 mL (P ¼ 0.0236) and 10,000 DPP/mL group (P ¼ 0.0159). Specific IgEs
and3.3againstgrasspollenandPhlp1,5,resp.)weretreatedwithanultra- remainedstableinallgroupswhereasspecificIgG1andIgG4showeddose-
rushschedulereceivingthemaximumdoseof0.2plus0.3mLofaallergen dependentincreases.Grade1systemicreactionsoccurredin18%(100DPP/
extract (DepiQuick, Novartis, Germany) on day one, followed by further mL), 14.8% (1000 DPP/mL), 17.4% (5000 DPP/mL) and 25.3% (10,000
injecting the maximum dose of 0.5 mL at weekly intervals for 5 weeks. DPP/mL)ofpatients.
Patientswereinterviewedbyaquestionnaire2monthsafterthepollenseason. Conclusions:Wedeterminedincreasedallergenamountstoobtainapositive
Results:75.8%ofpatientsreportedagoodorverygoodeffectofSITwith CPTafterSITfrom1000to10,000DPP/mLdepigmentedpolymerizedbirch
respecttotheirsymptomsduringthepollenseasonafterhavingreceivedonly pollen extract. The 5000 DPP/mL dose extract suggest a good benefit/risk
onecycleofultra-rushSIT;18.4didnotnoticeanyeffectafterthefirstcycle; ratiowiththepotentialforfurtherdevelopment.
onepatientreportedanincreaseofsymptoms.56.5%ofpatientsdiduseless
anti-allergicmedicationincomparisonwiththeirmeanneedbeforeSIT;in
18.8%theneedofmedicationwascomparabletotheyearbefore.In77.8%of
allcasesultra-rushSITwaswelltoleratedsubjectively.Localswellingatthe
injection site was reported in 25.9% (immediate) and 42.4% (delayed); 4
3
patients felt a mild discomfort after injection with pruritus, fatigue, or
Double-blindStudyoftheUseofTransferFactor(TF)Combined
dizziness, respectively. The systemic symptoms disappeared spontaneously
WithSublingualImmunotherapyinManagementofPatients
withoutmedication.
Conclusions:EvenforpatientswithclinicallysevereSARandhighspecific WithAllergicRhinitisinMexicanPopulation
sensitization against birch or grass pollen the pre-seasonal ultra-rush SIT MaríaIsabelRojoGutiérrez,1MisaelGonzalez-Ibarra,QFB,2TeresaSandoval,
regimen with depigmented, polymerized allergen extracts showed a good QFB,3andJaimeMellado-Abrego,MD1.1Allergy;2AllergyandImmunology;
efficacyandtolerability. 3Investigation,JuarezHospital,MexicoCity,Mexico.
Background: Allergic diseases affect20% of world population and allergic
rhinitis(AR)isthemostcommon,aloneorassociatedwithasthma.Antigen-
specificimmunotherapyhasbeenworld-desensitizingtypestudiedshowingthat
itiscapableofmodifyingthenaturalhistoryofthediseasethroughchangesin
2 theimmuneresponseincreasingtheroleofclonesoflymphocytesThelpertype
DoseOptimizingStudyofaDepigmentedPolymerizedAllergen I (LTh1) and inhibiting the activity of type 2 (LTh2). Dialyzable leukocyte
ExtractofBirchPollenbyMeansofConjunctival extractortransferfactor(TF)hasshownthesamestimulatoryeffect.
ProvocationTest Objective: Evaluate the synergistic effect of TF associated with SLIT in
Angelika Sager, MD,1 and Margitta Worm2. 1Medical Department, LETI patientswithRA.
PharmaGmbH,Witten,Germany;2Dermatology,ChariteUniversityClinic, Material and Methods: We studied 94 patients with RA. We included
Berlin,Germany. randomized into 2 groups (47 each one) both received antigen-specific
Background: Clinical efficacy of a depigmented polymerized birch pollen sublingualimmunotherapy(SLITAE)andalsoingroup1wasgivenoralTF
extracthasbeenshownin2largephaseIIIstudieswith1000DPP/mL.To Unitatone200mgmonthlyfor3months,andgroup2placebo.Weused
datedose-responsestudiesarerequiredtoshowoptimalefficacyatadefined acontrolgrouponlytocomparedthenormalresults.Allpatientsunderwent
allergendose.Theconjunctivalprovocationtest(CPT)isapossibleoutcome complete blood count (BHC), immunoglobulins, quantification of cell-type
parameterandfromtheregulatoryauthoritiesacceptedtestprocedure. lymphocyte CD3, CD4, CD8, interleukin (IL2, IL4, IL5, IL10, TNF and
WAOJournal (cid:2) February2012 S1
Abstracts WAOJournal(cid:2)February2012
IFN).Thenasalsymptomswereevaluatedmonthlyfor3months,withtest
scoresymptomsquestionary4(TSSQ4)andauto-evaluationbyscaleLinker. 5
Results:Westudied94ARpatients(50%womenand50%men,rangeage Posttreatment,Long-TermClinicalEfficacyofa300IR
between 5 and 40 years). Indoor antigens (mites, house dust and SublingualTabletof5-GrassPollenAllergenExtractinAdults
cockroach) were the main cause of allergy. Allergic patients had more WithGrassPollenInducedAllergicRhinoconjunctivitis
eosinophilesandIgEthanthehealthycontrols(P,0.05).Thenumberof
Alain Didier,1 Friedrich Horak,2 Margitta Worm,3 Hans-Jorgen Malling,4
CD81lymphocyteswasslightlyreducedingroup2aftertreatment(P,
Armelle Montagut,5 Patricia Rodriguez,5 Robert K. Zeldin,5
0.05), whereas the amount of IL-4 and IFN ƒnwere increased in both
and Michèle Lheritier-Barrand6. 1Respiratory Diseases Department, Ran-
groups(P,0.005)andthe amountofIL-10wassignificantlyincreased
gueil-Larrey Hospital, Toulouse, France; 2Allergy Centre Vienna West,
ingroup1(P,0.01)aftertreatment.Clinicalevaluationwaswithinitial
Dptm. - Institute for Allergy Research, Vienna, Austria; 3Allergy-Centre-
TSSQ4of11.6beforehandlingand5.1(44%)after,withsignificantim-
Charité, Charité - Universitätsmedizin Berlin, Berlin, Germany; 4National
provement(P,0.0001)andLikertscorewasreduced69%thanthestar
University Hospital, Copenhagen, Denmark; 5Stallergenes SA, Antony,
thetreatment.
France;6Stallergenes,AntonyCedex,France.
Conclusions:TheTFalongwithSLITAEinthetreatmentofpatientswith
Background: A 5-year study of adults with grass-pollen related rhinocon-
RAdidnotaltertheclinicalimprovementinducedbySLITAEalonefor3
junctivitishasdemonstratedthesustainedefficacyofdiscontinuoustreatment
monthsoftreatment,butthecombinationincreasedproductionofIL10and
witha300IRsublingualtabletof5-grasspollenallergenextract,initiated4or
productionofIFNg.
2 months before each pollen season and continued for its duration for 3
consecutiveyears.Herewereportonthepersistenceofefficacyduringthe
firstof2post-treatmentpollenseasons.
Methods:633adultswererandomizedtoeitherplacebooroneof2active
4 groupsreceivingpre-andco-seasonaltreatmentfor3pollenseasonsstarting
TheRiskofSevereTreatmentRelatedAdverseEventsis eachyeareither4months[4M]or2months[2M]priortothepollenseason.
SignificantlyLowerinChildrenComparedtoAdultsWhen Patients were followed during the subsequent, treatment-free, grass pollen
TreatedWithStandardizedTimothyGrassAllergy season.TheprimaryendpointfortheYear4assessmentofthepost-treatment
ImmunotherapyTablets:PostHocAnalysisof7ClinicalTrials long-term efficacy was the Average Adjusted Symptom Score (AAdSS,
JensStrodlAndersen,1PeterAstrupFejerskov,1andHendrikNolte2.1ALK- adjustingtheRhinoconjunctivitisTotalSymptomScoreforrescuemedication
Abello,Hørsholm,Denmark;2MerckResearchLaboratories,Kenilworth,NJ. usage)duringthefourthpollenperiod.Secondaryefficacycriteriaincluded
the Average Rescue Medication Score (ARMS) and the overall Rhinocon-
Background: Allergy immunotherapy tablets (AIT) provides a safer and
more convenient alternative to subcutaneous specific immunotherapy junctivitisQualityofLifeQuestionnaire(RQLQ)score.
Results: Statistically significant differences compared to Placebo in the
treatment. However, severe adverse events occur infrequently. These
meanAAdSSduringtheYear4pollenperiodwereobservedforboth300
eventsarerareandthereforepooledsafetydatafrom1phase II/IIIand6
phaseIIIclinicaltrials(5inadults,2inchildren(5–17years))withgrass IR (4M) and 300 IR (2M). The treatment effect for 300 IR (4M) was
estimated as the difference in LS Means of 21.14 (95% CI, [22.03 to
AIT(Grazax,Phleumpratense75,000SQ-T/2800BAU,ALK,Denmark)
20.26],P¼0.0114),correspondingtoarelativeLSMeandifferencefrom
wereanalysed.
Placeboof222.9%,whilstthetreatmenteffectfor300IR(2M)isestimated
Method: All trials were randomised, double-blind, placebo-controlled
asthedifferenceinLSMeansof21.43(95%CI,[22.32to20.53]),P¼
multi-centre trials. Subjects suffered from grass pollen induced allergic
0.0019),corresponding toarelativeLS Meandifference fromPlacebo of
rhinoconjunctivitiswithorwithoutasthma,hadpositiveskinpricktestand
specific IgE to Phleum pratense. Subjects received once-daily sublingual 228.5%.Theprimaryresultswereconfirmedovertheworstpollenperiod.
Compared to placebo, the active treatment groups (4M and 2M) also
treatment with grass AIT or placebo for approximately 24 weeks. The 5
adulttrialscomprised2095treatedsubjects(AIT¼1060,placebo¼1035) showedastatisticallysignificantLSMeandifferenceinARMS(224.6%;
andthe2childrentrialscomprised597treatedsubjects(AIT¼301,pla- P ¼ 0.00184 and 227.9%; P ¼ 0.0082) and in overall RQLQ score
cebo ¼ 296). Adverse events were assessed by the investigator as treat- (232.8%; P ¼ 0.0001 and 237.6%; P , 0.0001). No unexpected risk
wasidentifiedinthisstudy.
ment-related(possibleorprobable)orunlikelyrelatedandforseriousness.
Applicationsite-relatedeventsweredefinedasadverseeventsinrelationto Conclusions: The post-treatment, long-term efficacy of 300 IR sublingual
tabletsofgrasspollenallergenextractwasdemonstratedduringthefirstof2
theoralcavity.
post-treatment pollen seasons. This persistent improvement was clinically
Results:Intheadulttrials,71%ofAIT-treatedsubjectsreportedtreatment-
meaningfultopatients.
relatedadverseeventscomparedto24%forplacebo.Inthechildrentrialsthe
correspondingnumberswere63%forAITand27%forplacebo.OftheAIT-
treatedsubjects2children(0.7%)and32adults(3.0%)experiencedsevere
treatment-relatedevents.Theoddsforsevereeventswas4.7timeslowerin
childrencomparedtoadults(odds-ratiowith95%CI,0.22[0.025-0.85],P¼ 6
0.019).BothAIT-treatedchildren(0.7%)and18(1.7%)oftheAIT-treated EscherichiacoliHeat-LabileEnterotoxin(LTS61K)Modulates
adultsexperiencedseveretreatment-relatedeventsthatwereapplicationsite- DendriticCellFunctionandAttenuatesAirwayInflammationin
related.Theoddsforhavingsevererelatedapplicationsiteadverseeventswas MouseModelofAllergicAsthma
2.6timeslowerinchildrencomparedtoadults(odds-ratiowith95%CI,0.39 Jiu-yaoWang,MD,PhD,1Yu-ShenHsuHsu,PhD,2andI-PingLin,MSc3.
[0.04-1.63],P¼0.27,notstatisticallysignificant).Noserioustreatment-re- 1College of Medicne, National Cheng Kung University HospiTak, Tainan,
latedadverseeventswerereported. Taiwan;2DevelopmentCenterforBiotechnology,Taipei,Taiwan;3College
Conclusion: This pooled analysis of over 2000 subjects in 7 clinical ofMedicine,NationalChengKungUniversity,Tainan,Taiwan.
trialsshows thattheriskofexperiencingseveretreatment-relatedadverse Background: Escherichia coli heat-labile enterotoxin (LT) with different
eventswassignificantlylowerinchildrencomparedtoadultswhentreated mutant forms has been used as adjuvant for vaccines due to its ability to
with of Timothy grass allergy immunotherapy tablets. This analysis enhance immune response to specific antigen in vivo. We hypothesis that
provides evidence that Timothy grass AIT is an important and safe LTS61KorLTS61Kmixedwithdustmiteallergen,Derp,(LTS61K/Derp)
immunotherapy treatment option in children with grass pollen induced can modulate dendritic cells (DCs) s’ functions thus alleviate allergen-in-
rhinoconjunctivitis. ducedairwayinflammation.
S2 (cid:1)2012WorldAllergyOrganization
WAOJournal(cid:2)February2012 Abstracts
Methods: Two protocols (ie, preventive and therapeutic protocol) were theplacebogroup(P,0.0001).TotalQoLscorewasimprovedinverum
designed to evaluate the effects of LTS61K in Der p sensitized and patients(P¼0.0254).5.4%ofthepatientsintheverumgroupand4.0%of
challengedmousemodelofasthma. thepatientsintheplacebogroupdevelopedmildsystemicreactions.
Results: Both intranasal inoculations with LTS61K or LTS61K/Der p Conclusions: The results show that specific immunotherapy with a depig-
decreased allergen-induced airway inflammation and alleviated systemic mentedpolymerizedextractof2taxonomicallynon-relatedpollen(birchand
T 2-typeimmuneresponse.Inaddition,bronchoalveolarlavage(BAL)fluids grass) was effective and safe demonstrated by clinical and immunological
H
andserafromLTS61K/DerptreatedmicehavehigherconcentrationsofDer parameter.
p-specificIgAthanthoseofothergroups.Intheinvitrostudy,bonemarrow-
derived dendritic cells (BMDCs) and DC cell line, DC2.4 cells stimulated
withLTS61K/Derpbothsecretedpro-inflammationcytokinesIL-6andTNF-
a.Incontrast,afterLTS61Ktreatment,onlyBMDCsdecreasedproductionof 8
IL-6andTNF-aaswellasdecreasedmaturation.Furthermore,wefoundthat AllergenicCompositionofPolymerizedAllergenExtractsof
pre-treatment BMDC with LTS61K inhibited Der p-induced NF-kB trans- Betulaverrucosa,DermatophagoidesPteronyssinusand
locationwhichmightexplainthedelayedmaturationanddecreasedproduc- PhleumPratense
tions of IL-6 and TNF-a in LTS61K pre-treated BMDCs. Intratracheally Enrique Fernandez-Caldas, PhD,1,2 Barbara Cases, PhD,1 Jose Ignacio
adoptive transferred with LTS61K- or LTS61K/Der p-primed DC2.4 cells Tudela, BS,1 Eva Abel Fernandez, BS,1 Miguel Casanovas, MD, PhD,1
orBMDCSintoDerp-sensitizedmicedecreasedinflammatorycellsinfiltra- and Jose Luis Subiza, MD, PhD1. 1Research & Development, Inmunotek
tion and TH2-type chemokines in BAL fluids and alleviated airway SL,Madrid,Spain;2DivisionofAllergyandImmunology,UniversityofSouth
inflammation.
FloridaCollegeofMedicine,Tampa.
Conclusions:OurresultsshowthatLTS61KmayinfluenceDCsmaturation
Background: Allergoids have been successfully used in the treatment of
anditscytokineproduction.Ontheotherhands,LTS61K/Derpmayinduce respiratory allergic diseases.Theyare modified allergenextracts thatallow
more Der p-specific IgA production to decrease allergic T 2 cytokine
H theadministrationofhighallergendoses,duetotheirreducedIgEbinding
responses and alleviate airway inflammation in murine model of asthma. capacity.They maintain allergen-specific T-cell recognition. Since they are
These finding suggested that LTS61K may have clinical application as an
native allergen extracts that have been polymerized with glutaraldehyde,
immune-modulatoreffectonthediseasesofallergyandasthma. identificationoftheallergenicmoleculesrequiresmorecomplicatedmethods.
Theaimofthestudywastodeterminethequalitativecompositionofdifferent
polymerized extracts and investigate the presence of defined allergenic
ALLERGEN IMMUNOTHERAPY 2 moleculesusingMassspectrometry.
Methods:ProteomicanalysiswascarriedoutattheProteomicsFacilityofthe
HospitalNacionaldeParapléjicos(Toledo,Spain).Afterreductionandalkyl-
7 ation, proteins were digested with trypsin and the resulting peptides were
ComparisonofEfficacyandSafetyofaDipigmentedPolymerized cleaned using C18 SpinTips Sample Prep Kit; peptides were separated on
AllergenExtractofGrassandBirchwithPlaceboinPatientsWith anUltimatenano-LCsystemusingaMonolithicC18columnincombination
Type-1AllergicRhinoconjunctivitis with a precolumn for salt removal. Fractionation of the peptides was per-
formedwithaProbotmicrofractioncollectorandMSandMS/MSanalysisof
AngelikaSager,MD,1TiloBiedermann,2andOliverPfaar,MD3.1Medical
offlinespottedpeptidesampleswereperformedusingtheAppliedBiosystems
Department, LETI Pharma GmbH, Witten, Germany; 2Department of Der-
4800plusMALDITOF/TOFAnalyzermassspectrometer.ProteinPilotSoft-
matology, University of Tuebingen, Tuebingen, Germany; 3Center for Rhi-
wareV2.0.1andtheParagonalgorithmwereusedfortheidentificationofthe
nology and Allergology, Department of Otorhinolaryngology, University
proteins.EachMS/MSspectrumwassearchedagainsttheSwissProt2010_10
HospitalMannheim,Wiesbaden,Germany.
database,Uniprot-ViridiplantaedatabaseandUniprot_Betuladatabase.
Background: Thesafetyandefficacyofspecificimmunotherapy(SIT)with
Results:Analysisofthepeptidesrevealedthepresenceofnativeallergensin
depigmentedandpolymerizedallergenextractsofpolleniswelldocumentedin
thepolymerizedextracts:Derp1,Derp2,Derp3,Derp8andDerp11in
several clinical trials. We investigated efficacy and safety of an extract
D.pteronyssinus;Betv2,Betv6,Betv7andseveralBetv1isoformsinB.
containing 2 taxonomically non-related pollen species (birch and grasses) in
verrucosaandPhlp1,Phlp3,Phlp5,Phlp11andPhlp12inP.pratense
asubcutaneousimmunotherapyover2pollenseasonsinco-sensitizedallergic
allergoids.Inallcases,potentialallergenicproteinswerealsoidentified,in-
patientswithrhinitisand/orrhinoconjunctivitiswithorwithoutallergicasthma.
cluding ubiquitin, actin, Eenolase, fructose-bisphosphate aldolase, luminal-
Methods:269(ITT)patientswithconfirmedrhinitisand/orrhinoconjuncti-
bindingprotein(Heatshockprotein70),calmodulin,amongothers.
vitis were treated during 2009 and 2010 in Germany, Romania, Poland,
Conclusions:Thecharacterizationoftheallergeniccompositionofallergoids
Lithuania,andBulgaria.Foreachpatienta1-daybuild-upphaseapplying0.2
is possible using MS/MS analysis. The analysis confirms the presence of
mLand0.3mLof1000DPP/mLallergenextractwasapplied.Duringthe
native allergens in the allergoids. Mayor allergens are preserved during
remaining18-monthperiodmaintenance500DPPwereadministeredin4to6
polymerization.
weeksintervals.Patientswererandomisedtothetreatmentgroupsona2:1
basis (175 verum: 94 placebo). The main parameter in this study was the
combinedsymptomandmedicationscoreduringthebirchandgrasspollen
season 2010 over 7 weeks. Secondary parameters were symptom score,
medicationscore,IgE,IgG4aswellasqualityoflife. 9
Results: During the 2010 season a statistically significant difference (P ¼ AProteomicStyleApproachtoCharacterizeaGrassMixProduct
0.0385)wasobservedbetweentreatmentgroups:inpatientstreatedwiththe RevealsPotentialImmunotherapeuticBenefit
allergenextractthemediantimeweightedAUCofthecombinedsymptom Murray Skinner, PhD, Alan Bullimore, Nicola Swanc, MS, and
and medication score was 5.70, in patients treated with placebo7.07. This Wemimo Alawode, PhD. R & D, Allergy Therapeutics, Worthing, United
effectwaspredominantlyduetothereductionofsymptomscoreby21%over Kingdom.
placebo.Theintakeofrecuemedicationwasverylowduringbothseasons Background: Grass allergy immunotherapies often consist of a mix of
leadingonlytoa10%reduction(ns).BirchandphleumspecificIgEdidnot different grass extracts each containing several proteins of different
changeduringthecourseofthestudyinbothgroupswhereasrespectiveIgG4 physiochemicalproperties;howeverthesubtlecontributionsofeachprotein
levelsincreasedonlyintheverumgroupandremainednearlyunchangedin are difficult to elucidate. This study aimed to identify and characterise the
(cid:1)2012WorldAllergyOrganization S3
Abstracts WAOJournal(cid:2)February2012
group1and5allergensina13grassextractandtostandardisetheextraction
method. 11
Methods:Thegrasspollenswereextractedinisolationandpooledandalso EfficacyandTolerabilityofHDMInjectiveImmunotherapyWith
incombinationandanalysedusingavarietyoftechniquesincludingenzyme- MonomericAllergoid
linked immunosorbent assay (ELISA), liquid chromatography-mass spec-
Enrico Compalati, MD,1 Isabella Atzeni, MD,2 Sergio Cabras, MD,3
trometry(LC-MS)andsodiumdodecylsulphatepolyacrylamidegelelectro-
Paolo Fancello, MD,2 Giulio Gaspardini, MD,4 Rocco Longo, MD,5
phoresis(SDS-PAGE).
Vincenzo Patella, MD,6,7 and Giorgio Tore, MD8. 1Allergy & Respiratory
Results: Gold-staining and IgE immunoblotting revealed a high degree of
DiseasesClinic,UniversityofGenoa,Genova,Italy;2ServiziodiAllergolo-
homology of protein bands between the 13 species and the presence of
gia,Ospedale"SanGavinoMonreale",ASL6,Sanluri(CR),Italy;3Casadi
a densely stained doublet at 25 to 35 kD along with protein bands at
Cura "Madonna del Rimedio", Oristano, Italy; 4Divisione ORL, Ospedale,
approximately 12.5, 17 and 50 kD. The doublet from each grass species
Cagliari,Italy;5ServizioTerritorialediAllergologia,AziendaSanitariaPro-
demonstratedahighlevelofgroup1and5interspecieshomology.However,
vinciale,ViboValenzia,Italy;6DivisionofClinicalImmunologyandAllergy,
there were a number of bands unique to specific grasses consistent with
GeneralHospital,ASLSALERNO,Agropoli(SA),Italy;7U.O.diAllergolo-
evolutionarychangeandindicativethatagrassmiximmunotherapeuticcould
giaeImmunologiaClinica,OspedaleGenerale,Agropoli(SA),Italy;8Divi-
beconsideredbroadspectrum.
sioneORL,Ospedale"SantissimaTrinità",Cagliari,Italy.
Conclusions: SDS-PAGE and IgE immunoblotting showed all 13 grasses
Background:Subcutaneousimmunotherapy(SCIT)isaneffectivetreatmentof
share a high degree of homology particularly in terms of group 1 and 5
respiratoryallergyandcarbamylatedmonomericallergoids(monoids),byvirtue
allergens.IgEand IgGELISApotencieswereshown tobeindependentof
of their reduced IgE-binding activity, resulted clinically safe by sublingual
extractionmethod.
administration. Purpose of this study was to investigate the efficacy and
tolerability of immunotherapy with house dust mites (HDM) monoid admin-
isteredbyinjectiverouteinpatientswithallergicrhinoconjunctivitis(AR).
Methods: A preparation of 0.70 mL of 10 BU/mL containing modified
10 extract with 50% Dermatophagoides pteronyssinus and 50% Dermatopha-
SeasonalVersusSymptom-basedEvaluationofaDepigmented goidesfarinae(amountofmajorallergen:4mgofgroup1permilliliter)was
Grass-BirchAllergoid delivered monthly for 12 months, following a 5-week build-up induction
phase (0.10–0.20–0.30–0.50–0.70 mL), to 58 patients (60% males, mean
Oliver Pfaar, MD,1 Tilo Biedermann,2 and Angelika Sager, MD3. 1Center
age25.1612.7)sufferingfromARduetomitesforatleast2years,whereas
forRhinologyandAllergology,DepartmentofOtorhinolaryngology,Univer-
60patientswithsimilarbaselinecharacteristicswereobservedascontrols.All
sity Hospital Mannheim, Wiesbaden, Germany; 2Department of Dermatol-
patientswereallowedtoassumetraditionaldrugtherapyfortheircondition.
ogy, University Hospital, Tübingen,Germany; 3Medical Department, LETI
Attheendofthestudychangesfrombaselineinsymptomsscores,innumber
PharmaGmbH,Witten,Germany.
ofdayswithdrugassumption,inseverityofAR(accordingtoARIAclassi-
Background:Thesafetyandefficacyofspecificimmunotherapy(SIT)with
fication)werecomparedbetweenthe2groups;moreoveranoverallassess-
depigmentedandpolymerizedallergenextractsofpolleniswelldocumented
ment of clinical efficacy and tolerability was based on patients’ and
in several clinical trials. The results of such clinical studies are highly
physicians’judgements(unsatisfactory,mild,good,optimal).
dependentonthequantityandqualityofpollenexposureandtheirmeasure-
Results: In respect to baseline both groups showed, after 1 year, an
ments.Toidentifyapollen-independentefficacyassessmentwecomparedthe
improvement in symptoms score (P , 0.001) with a significant difference
combinedsymptomandmedicationscore(SMS)measuredduringthepollen
infavourofSCITgroup(P,0.05).Daysofdrugintakeweresignificantly
seasonwiththeSMSdefinedbyaminimumsymptomscoreoftheplacebo
lower in patientsreceiving SCIT(P , 0.05).The number of patients with
group in a subcutaneous immunotherapy with a depigmented Grass-Birch
severe AR decreasedinthe first groupwhile no variation wasobserved in
allergoidover2years.
controls.Thesubjectiveclinicaloverallassessmentwasoptimalin31cases
Methods:269(ITT)patientswithconfirmedrhinitisand/orrhinoconjuncti-
andgoodin24accordingtophysicians’andpatients’judgements;similarly
vitis were treated during 2009 and 2010 in Germany, Romania, Poland,
38patientsjudgedtolerabilityasoptimaland18asgood,whereasaccording
Lithuania,andBulgaria.Patientswererandomisedtothetreatmentgroupson
tophysiciansitwasoptimalin37patientsandgoodin19;inonly1patient
a2:1basis(175verum:94placebo).Themainparameterinthisstudywasthe
thetreatmentwasconsideredunsatisfactory.
combinedsymptomandmedicationscoreduringthebirchandgrasspollen
Conclusions:Inthisprospectivecontrolledstudy,SCITwithHDMcarbamy-
season20101.InadditiontheSMSoftheactivelytreatedpatientswasana-
lated allergoid was associated with a significant clinical benefit observed
lyzedfollowingtheplacebotreatedpatientselicitingaconsiderablesymptom
throughobjectiveandsubjectiveoutcomes;thetraditionalsafetyofmonomeric
burden.2.
allergoidswasconfirmedbythesubjectivejudgementsoftolerability.
Results:Theclinicalresultsfollowingtheseasonalapproacharegiveninthe
otherabstractofourgroup1.Takingalldayswithmeansymptomscore.2in
theplacebogroupascalculationbasis,thecombinedSMSvalueswerecon-
ALLERGEN STRUCTURE 1
siderablylowerforactivelytreatedpatientsthanforplacebotreatedpatients.
Forbothseasons,thedifferencesbetweenthetreatmentgroupswerehighly
statisticallysignificant(median;2009:5.06vs7.97,2010:4.26vs6.43;ITT
12
set)withP-valuesof0.0038and0.003.
Conclusions: The results show that the efficacy assessment of specific ComprehensiveDetectionofAllergensinGrassPollenExtractsby
MassSpectrometry
immunotherapy might be better discriminated in relation to the actual
symptoms of the placebo group rather than following the days of pollen Steffen Augustin, PhD,1 Liane Mitulski,2 Oliver Cromwell, PhD,3
exposure. Gerald Reese, PhD,4 and Andreas Nandy, PhD5. 1Allergopharma Joachim
Ganzer KG, Laboratory Molecular Biology, Reinbek, Germany; 2Allergo-
pharma Joachim Ganzer KG, Reinbek, Germany; 3Research and Develop-
REFERENCE ment; 4Allergen Research; 5Molecular Biology Research, Allergopharma
1. Abstract#3093:BiedermannT,PfaarO,SagerA."Comparisonofefficacy JoachimGanzerKG,Reinbek,Germany.
andsafetyofadepigmentedpolymerizedallergenextractofgrassandbirch Background: More than 40% of type 1-allergic individuals suffer from
withplaceboinpatientswithtype-Iallergicrhinoconjunctivitis.". hypersensitivitytograsspollen.Patientsaretreatedtraditionallywithspecific
S4 (cid:1)2012WorldAllergyOrganization
WAOJournal(cid:2)February2012 Abstracts
immunotherapy using pollen extracts derived from one or several different
Pooideae species. While for several species the most important allergens 14
(group1andgroup5)havebeenidentified,otherallergenshaveeithernot ComprehensiveExpressionofRecombinantHouseDustMite
beenidentifiedorsequencedataarestillmissing.Wehaveusedmassspec- AllergensforComponent-ResolvedDiagnosis
trometry(MS)togetherwithgeneticandimmunologicalmethodstoidentify
YoshiharuWatakabe,1SeijiKawamoto,1HikaruNakahara,1TakaakiHiragun,2
allergensinvariousgrasspollenextracts.
Tsunehiro Aki,1 Yoshiko Asaoku,3 Takaharu Hayashi,4 Shoji Mihara,2
Methods: Pollen extracts of 6 different grass species (Phleum pratense,
MichihiroHide,2andKazuhisaOno1.1DepartmentofMolecularBiotechnology,
Holcuslanatus,Loliumperenne,Dactylusglomerata,Festucapratensis,
GraduateSchoolofAdvancedScienceofMatter,and;2DivisionofMolecular
Poapratensis)andamixturethereofwereanalyzed.Foridentificationof
MedicalScience,DepartmentofDermatology,GraduateSchoolofBiomedical
allergensbyMS,extractsweresubjectedtoenzymaticdigestion.Result-
Sciences,HiroshimaUniversity,Hiroshima,Japan;3NagasakiHospital,Hir-
ing peptides were separated by liquid chromatography and analyzed by
oshima,Japan;4TakanobashiCentralHospital,Hiroshima,Japan.
tandem mass spectrometry. Protein identification was performed by
Background:Allergen-specificimmunotherapy(SIT)istheonlypromising
searchingboththeNCBIPlantreleaseandanindividuallydesigneddata-
treatmentofallergy.However,currentSIThaslimitationssuchasaneedfor
base. The presence of individual allergens was confirmed with allergen-
long-term medication and a risk of systemic anaphylaxis. Those issues are
specificmonoclonalantibodies.Unknownsequencesweredeterminedfol-
raised mainly because current SIT procedure is carried out using crude
lowingcDNAsynthesisfrompollenRNAandallergensequenceamplifi-
allergenextract,whichmayalsoinduceaharmfulneo-sensitization.Useof
cationbyPCR.
definedrecombinantallergenswouldbeapreferablealternativeforthenext
Results: Fes p 1 and Fes p 5 were identified by the PCR approach. MS
generationSITvaccineaswellasforthedevelopmentofcomponent-resolved
analysisofpollenextractsfromthe6individualspeciesresultedindetection
diagnosis (CRD), which enables to prescribe a patient-tailored vaccine.
ofallknownallergensincludingthenewlyidentifiedFesp1andFesp5.
Objective of this study is to construct a production system of recombinant
Basedonthehomologyofallergensfromdifferentgrassspecies,previously
house dust mite (Dermatophagoides farinae) allergens, and to test their
unknown sequences of representatives of groups 2, 3, 4, 7, 11, 12 and 13
usefulnessformoleculardiagnosis.
weredetectedbyMSininvestigatedextractswithhighsequencecoverage.
Methods:Thusfar,theWHO/IUISallergennomenclaturesubcommitteehas
Group6allergenscouldnotbeidentifiedinsomeoftheanalyzedextracts.
approved 24 Dermatophagoides allergens. Among them, we sought to
These findings are supported by immunological analyses and thus demon-
express 20 groups of D. farinae allergens (Der f 1, 2, 3, 4, 5, 6, 7, 8, 9,
stratethespecificityoftheappliedmethod.Membersofallallergengroups
10,11,13,14,15,16,17,18,20,21,and22)usingtheEscherichiacolicold
wereidentifiedinanextractmixpreparedfrompollenofall6grassspecies
shockexpressionsystem. We also tried to express additional new antigens
studied.
[Mag133 (a highly-conserved UK114/YER057c/YjgF family member),
Conclusions:Themostimportantgrassallergens(group1andgroup5)were
DFA22(anew group2 familymember),andDFA67(peroxiredoxin)]that
detectedinallextracts.Inadditionallotherknownallergensoftheassayed
weoriginallyidentifiedasmajorallergenswithhighIgE-bindingfrequencies.
speciesandhomologuesthereofcouldalsobeidentified,thusdemonstrating
IgE-bindingabilityofthoserecombinantallergenswasassessedbywestern
thequalityofthetestedextracts.
blotanalysis.Wealsotestedwhethertheseallergenswereapplicableforthe
developmentofCRD.
Results: We confirmed successful expression of above D. farinae allergen
molecules as soluble recombinant proteins. Western blot analysis revealed
13 that these recombinant allergens retained IgE-binding capacity. We also
GeneExpressionPatternofArabidopsisEXPB1,aNonallergenic foundthathousedustmite-allergicpatientsshoweddifferentialIgE-binding
HomologueofGrassGroup1PollenAllergens signaturesagainstthem,suggestingthatourrecombinantallergensareuseful
todeterminesensitizedallergenmoleculesinindividualpatients.
PremBhalla,PhD,RubyTiwari,PhD,andMohanSingh,PhD.Agriculture
Conclusions: Here we carried out the comprehensive expression of
andFoodSystems,TheUniversityofMelbourne,Parkville,Australia.
recombinantD.farinaemajorallergens.Therecombinantallergenrepertoire
Background: Grass pollen allergy is one of the most common allergies
offersanessentialplatformforthefuturemoleculardiagnosticsofdustmite
worldwide.GroupI allergensconstitute the majorallergeniccomponentof
allergy.
grasspollenwithmorethan85%ofgrasspollenallergicpatientsshowingIgE
reactivity. These are highly immunologically cross-reactive glycoproteins
specificallyexpressedinpollenofallgrasses.Alignmentsoftheamino-acid
sequencesofgrassgroupIallergensderivedfromdiversegrassspeciesreveal
upto95%homology.Itisthereforelikelythatthesemoleculesshareasimilar 15
biologicalfunction. ABioinformaticApproachtoAllergenNomenclatureAppliedto
Methods: RT-PCR analysis, In situ hybridisation, Promoter-GFP construct AllergensFromtheNon-BitingMidgeChironomus
design,planttransformationandanalysisoftransgenicplants. thummithummi
Results: Sequence comparison has identified a homologue (b-expansin Heimo Breiteneder, PhD,1 Peter Rozynekc, MSc,2 Monika Raulf-
clone At2g20750 or EXPB1) in Arabidopsis of the Cyn d 1 gene. The Heimsoth, PhD,2 Richard Goodman, PhD,3 and Christian Radauer, PhD1.
EXPB1proteinis42%similartotheCynd1protein.Thisgenerepresents 1DepartmentofPathophysiologyandAllergyResearch,MedicalUniversity
a member of a small multigene family in Arabidopsis. RT-PCR analysis ofVienna,Vienna,Austria;2InstituteofPreventionandOccupationalMed-
showedexpressiononlyinfloralnotvegetativetissues.Insituhybridisation icine of the German Social Accident Insurance, Ruhr-University Bochum,
using150bpregionofthe39UTRoftheArabidopsisgeneasprobeshowed Bochum,Germany;3FoodAllergyResearch&ResourceProgram,Univer-
specific expression in mature Arabidopsis pollen. We further cloned the sityofNebraska-Lincoln,Lincoln,NE.
promoterregionfortheArabidopsisEXPB1andpreparedandGFPfusion Background: Representatives of the family Chironomidae (non-biting
constructs.TheseconstructswerethenintroducedintoArabidopsisplants midges;orderDiptera)arefoundworldwide.Freeze-driedchironomidlarvae,
byfloraldipmethod.GFP-promoterfusionsshowedhighlevelofexpres- predominantlyofthespeciesChironomusthummithummiarefrequentlyused
sionintri-cellularpollen. asfishfoodandareanallergensourceforfishkeepersandpersonsemployed
Conclusions: Our study provides evidence that EXPB1, a non-allergenic inthemanufactureoffishfood.Atpresent,9allergensofC.thummithummi
homologue of grass group 1 pollen allergens, gene is expressed in mature have been assigned an official designation by the WHO/IUIS allergen no-
pollen. menclaturesub-committee:Chit1to9.Allofthemarehemoglobinswith
(cid:1)2012WorldAllergyOrganization S5
Abstracts WAOJournal(cid:2)February2012
molecular weights of 16 kDa. IgE binding and cross-reactivity was clearly
demonstrated for all these proteins. However, the assignment of 9 distinct 17
allergennumberstomembersofthesameproteinfamilyisquiteunusual. SensitizationtoBetulaverrucosaandRBETV1inSpain
Methods: Currently, the IUIS allergen database contains 12 allergen and
Barbara Cases, PhD,1 Ramon Orjales,MD,2 Guro Gafvelin,PhD,3 Francisco
isoallergen sequences from C. thummi thummi. The Uniprot database has
Carballada,MD,PhD,2ManuelBoquete,MD,PhD,2JoseIgnacioTudela,BS,1
demergedentryP02225,listedinthedatabaseforChit7,into7entries,5
andEnriqueFernandez-Caldas,PhD,1,4.1Research&Development,Inmunotek
fromC.thummithummiand2fromC.thummipigerthatareidenticalto2of
SL, Madrid, Spain; 2Servicio de Alergia, Hospital Xeral-Calde, Lugo, Spain;
thesequencesfromC.thummithummi.Consequently,the16uniqueamino
3KarolinskaInstitut,Stockholm,Sweden;4DivisionofAllergyandImmunology,
acidsequencesofthematureC.thummiallergenswerealignedusingClus-
UniversityofSouthFloridaCollegeofMedicine,Tampa,FL.
talX2,aneighbor-joiningtreewasgeneratedfromthealignmentandapercent
Background:SensitizationtoBetulaverrucosapolleniscommoninNorth
sequenceidentitymatrixwasbuilttoevaluateappropriatenomenclature.
CentralEurope.InSpain,itisalsocommoninpatientsfromGalicia,aregion
Results: Pairwise sequence alignments showed that sequences belonging to
locatedintheNorthwestofSpain.Birchtreesareabundantinthisregionand
allergensChit5,6,7and8possesssequenceidentitiestoChit3ofbetween51
severalbirchspecieshavebeendescribed.Themainobjectivesweretode-
and63%.Chit1,2,4,and9divergetoagreaterextentfromChit3(,50%
termine the differences in the IgE reactivity to B. verrucosa and rBet v 1
identical)andfromeachother.Phylogenetictreeanalysissuggeststheclustering
betweenpatientsfromSwedenandSpain,aswellasthecontributionofthe
ofChit3,6,7,and8,whileChit1,2,4,5,and9formseparateclades.
majorallergenBetv1totheoverallreactivityofB.verrucosa.Allergenicity
Conclusions:Basedontheseanalyses,theIUISAllergenNomenclatureSub-
wasalsocomparedusingaserumpoolderivedfromseraofNorthAmerican
CommitteerenamesChit5,6,7and8isoallergensofChit3,eventhough
(USA)patients.
theirsequenceidentitiestoChit3arebelowthe67%thresholdpreviously
Methods:IgEreactivitytoB.verrucosaandrBetv1wasmeasuredinsera
definedforisoallergens.Theremaininghemoglobins,previouslydesignated
from 44 Spanish and 21 Swedish patients. rBet v 1 was produced as N-
Chit1,2,4and9willretaintheirpreviousnames.
terminalHis-tagged fusionproteinand purified as originally described. All
ofthemweresensitizedtothepollenofthistreespeciesandsufferedfrom
allergicrhinitisduringthepollinationperiod.rBetv1contributionwasde-
termined by inhibition ELISA using 3 pools of sera: from Spanish, North
16 AmericanandSwedishpatients.IgEbindingpatternwasevaluatedbyWest-
PollenAllergensDifferFromNonallergenicPollenProteinsby ernblotsusingthesamepoolsofsera.SpecificIgEbindingwasexpressedin
TheirLowerExtentofEvolutionaryConservation arbitraryunits.
Results: Specific IgEtoB. verrucosawasdetectedin50fromthe65 sera
Christian Radauer,PhD, Eva Guhslc,MSc, Merima Bublin,PhD, and Heimo
analyzed(mean¼3.7763.71Units).SpecificIgElevelstorBetv1were
Breiteneder,PhD.DepartmentofPathophysiologyandAllergyResearch,Medical
3.1363.32Units.ImmunoblotassaysconfirmedspecificIgEbindingtorBet
UniversityofVienna,Vienna,Austria.
v 1, and also to other allergens present in the extract. The Spanish pool
Background:Pollencontainshundredsofdifferentproteins.However,only
asmallfractionofthemhavebeenidentifiedtobeallergenic.Weaimedto presentedreactivitytomoreallergensthantheSwedishpool.ELISAInhibi-
tionassays,performedwithanativeextractandrBetv1,revealedasignificant
testthehypothesisthatmostpollenproteinsarenon-allergenicduetotheir
contribution(.80%inhibition)ofrBetv1totheallergenicityoftheextract,
highextentofsequenceconservationamongnon-relatedspecies.
withnodifferencesaccordingtotheoriginofthesera.
Methods: Data on the composition of pollen proteomes of birch (Betula
Conclusions:Betv1hasagreatimportanceinbirchallergyinGaliciaand
pendula), pellitory (Parientaria judaica) and timothy grass (Phleum pra-
Sweden. Nevertheless, there are differences in the IgE recognition pattern
tense)wereobtainedfromtheliterature.Sequencesweredownloadedfrom
UniProtandmanuallyclassifiedintoallergensandnon-allergens.Complete accordingtotherestofbirchallergens.Mayorallergenfrombirch,Betv1,
significantlycontributestotheallergenicityofB.verrucosainGalicia.
proteomesequencesof3dicotyledonousspecies(Arabidopsisthaliana,Pop-
ulustrichocarpaandVitisvinifera),2monocotyledons(Oryzasativasubsp.
japonicaandZeamays)andonemoss(Physcomitrellapatents)weredown-
loadedfromENSEMBLPlants.Sequencesofpollenproteinswerecompared
to these proteomes by using BLAST and the hits yielding the highest se-
ALLERGEN STRUCTURE 2
quence identity recorded taking into account only sequence alignments at
least40residuesinlength.Thedistributionsofmaximumsequenceidentities
of allergensand non-allergens from each species were compared using the
Mann-Whitneytest. 18
Results: Allergens from birch and pellitory pollen were significantly (P , ProteomicandImmunologicalCharacterizationof
0.001)lesssimilartoproteinsfrommonocotsthannon-allergenicpollenpro- RagweedAllergens
teins. Median sequence identities to the nearest rice and maize homologues Steffen Augustin, PhD,1 Marion Stock,2 Oliver Cromwell, PhD,3
were49and52%forbirchallergens,86and85%forbirchnon-allergens,37 Andreas Nandy, PhD,4 and Gerald Reese, PhD5. 1Allergopharma Joachim
and37%forpellitoryallergens,and87and89%forpellitorynon-allergens. Ganzer KG, Laboratory Molecular Biology, Reinbek, Germany; 2Allergo-
Similarly,timothygrasspollenallergensweresignificantly(P,0.0001)less pharma Joachim Ganzer KG, Reinbek, Germany; 3Research and Develop-
similartodicotproteinsthannon-allergenicpollenproteins.Mediansequence ment; 4Molecular Biology Research; 5Allergen Research, Allergopharma
identitiestothenearesthomologueswere43to44%forallergensand81to JoachimGanzerKG,Reinbek,Germany.
83%fornon-allergens.Acomparisonofall3pollenproteomestosequences Background:TheprevalenceofsensitizationtoragweedhasriseninNorth
fromthemossP.patensyieldedsimilarlysignificantdifferences. AmericaandacrossEurope.AlthoughthepectatelyaseAmba1,themajor
Conclusions: Pollen allergens belong to evolutionary less conserved protein allergenofragweed,wasidentifiedaslongagoasthe1960s,littleisknown
families than non-allergenic pollen proteins. The continual exposure of the about the allergenicity of the 5 Amb a 1 isoallergens and other allergens
human immune system to nearly identical and hence highly cross-reactive present in ragweed pollen. Ragweed extracts and purified Amb a 1
conservedproteinsfrommultiplepollenandplantfoodspeciesmostlikelyleads isoallergens have now been characterized for their allergenic potential to
totheinductionofimmunologicaltoleranceratherthanallergicsensitization. determine whether a single Amb a 1 isoallergen, several isoallergens or
This study was supported by grants P-22559-B11 (to CR) and SFB-F01802 acombinationwithotherallergensshouldbeincludedinarecombinantSIT
(toHB)fromtheAustrianScienceFund. vaccine.
S6 (cid:1)2012WorldAllergyOrganization
WAOJournal(cid:2)February2012 Abstracts
Methods:ExtractsfromNorth Americanshort ragweed(Ambrosia artemi-
siifolia)pollenwereinvestigatedbymassspectrometry(MS),2D-PAGEand 20
immunoblotting. Furthermore, Amb a 1 isoallergens were purified and IgE Cross-ReactivityBetweenOlivePollenand3SpeciesofGrassesin
reactivitydeterminedbyimmunoblottingandIgEinhibition. Madrid,Spain
Results:2D-PAGEandMSofragweedextractprovedthepresenceofall5
Enrique Fernandez-Caldas, PhD,1 M. Dolores Ibañez, MD,2 Barbara
knownAmba1isoallergens,ofwhichAmba1.01representsthedominant
Cases, PhD,1 Eva Abel Fernandez, BS,1 Jose Ignacio Tudela, BS,1
form.Additionallyallotherragweedallergensknownbysequence(Amba3,
Silvia Sanchez-Garcia, MD,2 Pablo Rodriguez del Rio, MD,2 and Miguel
Amba4,Amba5,Amba6,Amba8,Amba9,Amba10)wereidentified.
Casanovas, MD,PhD1. 1Research & Development, InmunotekSL, Madrid,
ThehighestIgEreactivitybyimmunoblottingwasobservedforAmba1.01
Spain;2AllergySection,HospitalInfantilUniversitarioNiñoJesús,Madrid,
followedbyAmba1.03;otherAmba1isoallergensaswellasotherdetected
Spain.
ragweedallergensshowedonlyweakIgEreactivity.Allisoallergenswiththe
Background:Themostcommonallergenicpollensinpatientswithpollinosis
exceptionofAmba1.04,whichisonlyoflowabundanceinragweedextract,
inCentralSpainaregrassesandolivepollen,withaprevalenceofpositive
were purified. Similar to the immunoblot analysis with crude extract, the
skinpricktestresultsof94and61%,respectively.Sharedproteinshavebeen
purifiedisoallergensAmba1.02andAmba1.05showedweakIgEbinding,
described in olive and grasses, such as profilins, polcalcin and trypsin
whereas Amb a 1.01 and Amb a 1.03 had high IgE reactivity. First IgE
inhibitors, but there are few in vitro studies analyzing the potential cross
inhibition experiments suggest that Amb a 1.01 contains all relevant IgE
reactivitybetweenbothspecies.Theaimofthepresentstudywastoanalyze
epitopes.
the protein composition of both allergen extracts, and the allergenic cross-
Conclusions: Amb a 1.01 is the most abundant Amb a 1 isoallergen, and
reactivitybetweenO.europaeaanddifferentgrasspollenspecies.
presumablythemostimportantragweedallergen.However,alargerpanelof
Methods:Seventy-two(72)patients(meanage10.4years)wereincludedin
ragweed-allergic subjectshastobe analyzedwithregardto IgEandT cell
the study. All of them suffered rhinoconjuntivitis and/or asthma and were
reactivities,tobeabletochooseacandidateforarecombinantvaccinefor
sensitizedtooliveand/orgrasspollen.SpecificIgEoftheindividualpatients
specificimmunotherapyofragweedallergy.
againstO. europaea,and tothe grass species:Dactylis glomerata, Phleum
pratense and Trisetum paniceum were determined by ELISA. Inhibition
assayswereperformedtoverifyallergeniccrossreactivitybetweengrassspe-
ciesandolive.Massspectrometryanalysiswasperformedtocharacterizethe
19 extractsandestablishiftherearecommonproteinsinboth,grassandolive
ProteomicAnalysisofMajorandMinorAllergensFromIsolated pollens,thatcouldactascrossreactiveproteins.
PollenCytoplasmicGranules Results:Threedifferentsensitizationpatternswereobserved:1)sensitization
OussamaAbouChakra,PhD,1,2Jean-PierreSutra,PhD,1EmmanuelleDemey tooliveandgrasspollen,2)sensitizationtooliveandnottograssesand3)
Thomas,MSc,3JoëlleVinh,PhD,3PascalPoncet,PhD,1,4GhislaineLacroix, sensitizationtograssesandnottoolive.Differentpoolsofseraweremixed
PhD,5 and Hélène Sénéchal, PhD,1,6. 1LSABM - Allergy and Environment, accordingtothisclassificationandusedforthedifferentassays.Correlation
ESPCIParisTech,Paris,France;2R&D,FAREVACARE,SavignyleTemple, coefficients found for the 3 grass species were significant (P , 0.0001;
France;3SMBP-CNRSUSR3149,ESPCIParisTech,Paris,France;4Infec- Spearman),butnotforolivepollen(P¼0.14;Spearman).Proteomicanalysis
tionandEpidemiology,InstitutPasteur,Paris,France;5DRC-TOXI,INERIS, revealedthepresenceofmorethan40commonproteinsingrassesandolive
Verneuil-en-Halatte,France;6CSS5,INSERM,Paris,France. pollens,butinhibitionassaysdemonstratednoallergeniccross-reactivitybe-
tweenbothfamilies.
Background: Grass pollen is one of the most important vectors of
Conclusions:ThereisnoinvitrocrossreactivitybetweenO.europaeaand
aeroallergens.Underatmosphericconditions,pollengrainscanreleasepollen
Grass pollen extracts, in spite of the allergens and the large number of
cytoplasmic granules (PCGs). The allergens associated with these intrinsic
commonproteinssharedbythesepollens.Wecanconcludethatsensitization
sub-fractions induce, in laboratory animals as well as in asthmatic patients
allergic and inflammatory responses. The aims of this study were to tooliveandgrassesisspeciesspecific.
characterize and identify the intrinsic allergens of PCGs, to compare them
withthoseofpollengrains.
Methods:PCGswereisolatedfromPhleumpratensepollenbyosmotic
shock.Thewater-solubleproteinswereextractedfrompollengrainsand
theirPCGs.Nineoutof26grasssensitizedpatientserawereselectedon 21
the basis of previous ELISA and immunoblotting results showing IgE MolecularPropertiesandImmunologicalReactivityof
specific binding to numerous grass pollen allergens. IgE-binding pro- ArabidopsisEXPB1,aNonallergenicHomologueofGrassGroup
teins were analyzed by 1- and 2D-immunobloting using grass pollen- 1Allergens
sensitized patient sera. Once located, allergens were characterized by MohanSingh,PhD,RubyTiwari,PhD,andPremBhalla,PhD.Agriculture
massspectrometry. andFoodSystems,TheUniversityofMelbourne,Parkville,Australia.
Results: 2D gels of pollen and PCGs extract revealed about 100 and 40 Background:Grassgroup1allergensareglyco-proteinsofabout30kDathat
proteins respectively, with a large spectrum of Mr (10–.94 kDa) and pI are highly soluble and profusely released by grass pollen upon hydration.
(,4.5–10.0). More proteins as well as more allergens in pollen than in They bind to IgE antibodies that initiate the allergic response causing hay
PCGsweredetectedbyimmunoblotting.Severaloftheallergenslistedin fever,seasonalasthma,andrelatedimmuneresponsesinhumans.Bermuda
theIUISnomenclature-Phlp1,4,5,6,11and12-werefoundinpollen grass(Cynodondactylon;subfamilyChloridoideae)isanimportantsourceof
andPCGsextractswhilePhlp11wasfoundonlyinPCGsandPhlp2as seasonal aeroallergens in warm tropical and sub-tropical areas worldwide.
wellasPhlp13onlyinpollenextract.Someotherallergens,notlistedin Improved approaches to diagnosis and therapy of allergic diseases require
the IUIS nomenclature, were also characterized in both pollen and PCGs athoroughunderstandingofthestructureandepitopesontheallergenmol-
extracts. eculethatarecrucialfortheantigen-antibodyinteraction.Inordertounder-
Conclusions:Sincethe major grasspollen allergens were foundinPCGs standstructuralbasisofIgEreactivityofgroup1allergenCynd1,wehave
and because of their small size, these sub-micronic particles should be pursuedacomparativegenomicapproachtosearchforhypoallergenicornon-
consideredasverypotentsensitizingandchallengingrespirablevectorsof allergenichomologues.
allergens.WedemonstrateherethatPCGsareatleastasmuchdangerousas Methods: Gene cloning, Protein expression in bacteria, protein structure
pollengrains. modeling,IgEreactivityanalysisthroughImmunoblotting.
(cid:1)2012WorldAllergyOrganization S7
Abstracts WAOJournal(cid:2)February2012
Results: EXPB1, an Arabidopsis protein (belonging to the beta expansin
multigenefamily),showedsignificantsequenceandstructuralsimilarityto 23
Cynd1.ThisproteinwasexpressedinE.coliandtherecombinantprotein GraftingofBETV1EpitopesontoitsHomologueAPIG1Reveals
didnotreactwithserumIgEfromgrasspollenallergicpatients,suggesting Patient-SpecificIgERecognitionProfiles
thatEXPB1representedanon-allergenichomologueofgrassgroup1aller-
BarbaraGepp,1NinaBalazs,1WolfgangHemmer,2ChristianRadauer,PhD,1
gens.Itisproposedthatdifferencesintheaminoacidsequencearerespon-
and Heimo Breiteneder,PhD1. 1Department of Pathophysiology and Allergy
sibleforthedifferenceintheallergenicityprofileoftheArabidopsisandgrass
Research,MedicalUniversityofVienna,Vienna,Austria;2FloridsdorferAller-
pollenproteins.
giezentrum,Vienna,Austria.
Conclusions:Ourstudyprovidesvaluabledataforfurtherinvestigationsof
Background:Upto70%ofbirchpollen-allergicindividualsshowadverse
the molecular basis of allergenicity and cross-reactivity of grass group 1
reactionstocertainplantfoods.Thiscross-reactivityiscausedbysensitisation
allergens.
tothemajorbirchpollenallergenBetv1andbindingofBetv1-specificIgE
antibodies to homologous plant food allergens. We aimed to assess the
importanceofselectedconformationalepitopesforIgEbindingtoBetv1.
Methods: Chimeras of Bet v 1.0101 and its homologue Api g 1.0101 were
constructed.Ineachofthe4chimeras,roughlyonefourthofthesurfaceresidues
22 ofApig1.0101werereplacedbycorrespondingresiduesofBetv1.0101.The
Protein-ProteinInteractionsDetermineIgEReactivityto proteins were expressed in Escherichia coli and purified by chromatographic
PolygalacturonaseFromCupressussempervirensPollen methods.SecondarystructureswerecheckedbyCD-spectroscopy.IgEELISA
Youcef Shahali,1 Jean-Pierre Sutra, PhD,1 Sylvie Chollet-Martin,2 withBetv1.0101,Apig1.0101andthechimeraswereperformedwithseraof63
Iman Haddad,3 Joëlle Vinh, PhD,3 Adriano Mari, MD,4 Denis Charpin,5 Betv1-sensitizedbirchpollenallergicpatients.ForinhibitionELISAs,chimeras
HélèneSénéchal,PhD,1,6andPascalPoncet,PhD,1,7.1UMR7195LSABM, werecoatedandinhibitionwasperformedwiththechimerasorApig1.0101.
ESPCI-ParisTech,Paris,France;2UnityofImmunologyAutoimmunityand Results:IgEbindingtoApig1.0101,Api-Bet-1,-2,-3and-4wasobservedfor
Hypersensibility,BichatHospital,Paris,France;3USR3149SMBP,ESPCI- 22,81,79,70and38%ofthesera,respectively.Toassesstherelevanceofthe
ParisTech,Paris,France;4CMA,Rome,Italy;5HopitalNorddeMarseille, graftedregionsforIgEbindingtoBetv1,theamountsofIgEbindingtothe
Marseille, France; 6CSS 5, INSERM,Paris, France;7Infection and Epide- chimeraswerecomparedwiththosetoApig1.0101.Mostoftheserarecognised
either3chimeras(39%)orall4chimeras(21%)betterthanApig1.0101.Only
miology,InstitutPasteur,Paris,France.
Background:Inarecentproteomicstudy,weidentifiedinItaliancypress aminorityoftheserashowedincreasedbindingtoasinglechimera.Inhibition
ELISAsconfirmedthepresenceofIgEspecificforthegraftedregions.
(Cupressus sempervirens, Cups) pollen grains, 2 proteins at 43 and 60
Conclusions:OurstudyindicatesthattheepitoperecognitionprofileofBetv
kDa, homologous to already known Cupressaceae polygalacturonase
1-specific IgE is highly patient specific. Due to the different IgE binding
(PG)proteins.The60-kDaPGissuspectedtobeamulti-proteincomplex
patternstoBetv1,determinedbybindingofIgEtodifferentchimeras,the
including the 43-kDa PG and one or more proteins with lectin-like
existence of a single major IgE epitope on Bet v 1 can be excluded.
properties
Moreover, the Bet v 1-specific IgE repertoire is polyclonal and the IgE
Objective:Inthepresentstudy,cypresspollenPGswerefurthercharacter-
epitopesaredistributedoverthewholesurfaceofBetv1.
izedandthemolecularbasisoftheirallergenicityincludingthepresenceof
specific IgE directed against cross-reactive carbohydrate determinants ThisstudywassupportedbygrantsP22559-B11(CR)andSFB-F01802(HB)
fromtheAustrianScienceFund.
(CCDs)wereinvestigated.
Methods:CupspollenPBSextractswerecharacterizedusing2-anddou-
bleone-dimensionalelectrophoresisfollowedbyIgEimmunoblotting.The
ALLERGIC MODELS OF INFLAMMATION
IgE reactivity to carbohydrate- versus peptide-specific determinants was
investigated using both bromelain inhibition and Con A-binding assays.
Pollen proteins were also prefractionated in their native forms using
sizeexclusionchromatography.Thepresenceofmulti-proteincomplexes 24
were investigated by using 2-D blue native (BN)-PAGE/SDS-PAGE CommonandRareVariationintheTHelper2GenePathway
electrophoresis. PredictsAllergicAsthmaPhenotypes
Results:Uponbromelaininhibitionassay,werevealedthat70%oftested Rebecca Slager, PhD, MS,1 Wendy Moore, MD,2 Huashi Li, MS,2
patientsdisplayedCCD-specificIgEtothe43-kDaPGwhileitsisoenzyme William Busse, MD,3 Mario Castro, MD, MPH,4 Serpil Erzurum, MD,5
of 60 kDa appeared to be exclusively recognized for its peptide-specific Anne Fitzpatrick, PhD,6 Sally Wenzel, MD,7 Deborah Meyers, PhD,1
determinants.ThespecificbindingoftheConAlectintothe43-kDaPG, and Eugene R. Bleecker, MD1. 1Center for Genomics and Personalized
and not to the60-kDa isoenzyme,demonstrated the presenceof exposed MedicineResearch,WakeForestUniversityHealthSciences,Winston-Salem,
mannose-containingoligosaccharidesonlyonthe43-kDaprotein.Thisfact NC;2CenterforGenomicsandPersonalizedMedicineResearch,WakeFor-
reflects fundamental differences between specific IgE-binding epitopes estUniversitySchoolofMedicine,WinstonSalem,NC;3Medicine,University
involved in the recognition of the 43-kDa and 60-kDa proteins making ofWisconsinSchoolofMedicineandPublicHealth,Madison,WI;4Division
these2cypresspollenPGsimmunologicallydistinguishable.Thepresent ofPulmonaryandCriticalCareMedicine,WashingtonUniversitySchoolof
results suggest that in the 60-kDa protein complex, the CCDs of the 43- Medicine, St. Louis, MO; 5Department of Pathobiology, Lerner Research
kDa PG are not exposed due to the binding of a lectin-like protein Institute, Cleveland, OH; 6Department of Pediatrics, Emory University,
exhibiting peptidic IgE reactive epitopes recognized by 25% of tested Atlanta,GA;7Medicine,UniversityofPittsburgh,Pittsburgh,PA.
patients. Background: The T helper 2 (Th2) inflammatory pathway, including the
Conclusion:ThecurrentstudydemonstratesthatthesensitizationtotheCups Th2-activatingcytokineinterleukin33anditsreceptorinterleukin1receptor-
pollenPGismainlyduetoCCDbromelain-typeepitopesanddirectlyasso- like1havebeenstronglyimplicatedinasthmasusceptibility(MoffattMF,et
ciated with an increased prevalence of IgE reactivity to cypress pollen al NEJM 2010). However, the role of Th2 pathway genetic variation in
extracts due to CCD interference. However, the Cups pollen PG and its asthmaprogressionandseverityisnotwellunderstood.Ourresearchgroup
carbohydrate-specificdeterminantsseemtoplayakeyroleinthedynamics recentlydevelopedaclusteringalgorithmbasedoncomprehensivephenotype
of protein-protein interaction in cypress pollen and may confer to protein informationtoassignsubjectswithasthmainthe SevereAsthmaResearch
complexesahigherallergenicity. Program(SARP)to5primaryclusters;3ofwhichrepresentincreasingsevere
S8 (cid:1)2012WorldAllergyOrganization
WAOJournal(cid:2)February2012 Abstracts
allergic asthma (Moore WC, et al AJRCCM, 2010). We hypothesized that
commonandpotentiallydeleteriousrarevariationinthispathwaywouldbe 26
associatedwithsevereasthmabasedonSARPclusterdesignation. MIR-150SuppressesLungInflammationinaMouseModelof
Methods: To evaluate common variants (minor allele frequency or MAF ExperimentalAsthma
.5%),419SARPnon-Hispanicwhiteparticipantswithaclusterassignment
Jia-WangWang,PhD,1KunyuLi,BS,1GaryHellermann,PhD,1RichardF.
weregenotypedfor182singlenucleotidepolymorphisms(SNPs)inTh2path-
Lockey, MD,2 Subhra Mohapatra, PhD,3 and Shyam Mohapatra, PhD4.
waygenesusingwhole-genomeSNPdata.IndividualSNPsandacumulative
1Internal Medicine, University of South Florida, Tampa, FL; 2Division of
modelofsignificantSNPswereevaluatedusingcontingencytableswithachi-
Allergy & Immunology, University of South Florida and James A. Haley
squaretestfortrendandordinalregressionmodelsadjustedforage,sex,and
Veterans’Hospital,Tampa,FL;3NanomedicineResearchCenterandDivi-
principalcomponents.Rare(MAF,5%)aminoacidchangesandsplicesite
son of Translational Medicine, Departments of Molecular Medicine and
alterations in this pathway were tested for association with asthma severity
InternalMedicine,UniversityofSouthFloridaandJamesVHaleyVeterans’
outcomesin20SARPsubjectswithwholeexomesequencedata.
Hospital, Tampa, FL; 4USF Nanomedicine Research Center, University of
Results:IndividualTh2pathwayvariantswereassociatedwithoverallSARP
SouthFloridaCollegeofMedicine&VAHospital,Tampa,FL.
clusterassignment,andallergicclustersofincreasingseverity(1,2,and4),
Background:Asthmaisacomplexdisorderoftheimmunesystemcausedby
including GATA3 polymorphism rs1244186 (P ¼ 0.005). In an 18-SNP
acombinationofgeneticpredispositionwithenvironmentalexposures.The
additivemodel,anincreasingnumberofTh2pathwayriskgenotypeswere
environmentalfactorsplayapredominantroleintheetiologyofasthma.Itis
highlyassociatedwithsevereallergicasthma(P¼3.9·1026).Forexample,
hypothesized that epigenetic changes in miRNAs play a critical role in
incluster4,thepercentageofsubjectswithatleast9riskgenotypeswas83%
pathogenesis of asthma as an interface between genetic makeup and
comparedto35%incluster1.Additionally,therewasevidencethatsubjects
environmental exposures. (Wang, Jia-wang; Li, Kunyu; Hellermann, Gary;
withrarevariantsinthispathwayweremorelikelytoreportallergysymp-
Lockey,RichardF.;Mohapatra,Subhra;andMohapatra,Shyam.Regulating
toms(P¼0.006),especiallyinthefall(P¼0.003),comparedtosubjects
theRegulators:microRNAandAsthma.WorldAllergyOrganizationJournal.
withnorarevariants.
June2011,Volume4,Issue6).
Conclusions:CommonTh2pathwayvariantspredictanincreasedlikelihood
Methods:Inthepresentstudy,weusedmiRNAarrayprofilinginamouse
of severe allergic asthma and rare variants were associated with increased
model of ovalbumin-induced asthma to identify differentially regulated
seasonalallergysymptoms.
miRNAs and characterized miR-150 in terms of cellular and humoral
involvementandanalysisoflunginflammationmarkers.
Results:WefoundthatmiR-150wasdownregulatedinCD4Tlymphocytes
duringasthmaticinflammationandTh1andTh2induction.Over-expressionof
25 miR-150deliveredbychitosannanoparticlesinhibitedlunginflammationand
RoleofMyeloidDerivedSuppressorCellsinAsthma decreasedTh1andTh2cytokinelevels.miR-150suppressedAkt3,Cbl1and
Allison Nelson,1 Jim Parkerson, DO,2 Richard F. Lockey, MD,2 Subhra Elk1oncogenes,whichareinvolvedininflammationandcytokineproduction.
Mohapatra, PhD,3 Shyam Mohapatra, PhD,4 and Srinivas Nagaraj, PhD1. Transgenic mice overexpressing miR-150 are resistant to asthma induction,
1Nanomedicine Research Center and Divisions of Translational Medicine demonstratedbyreducedAHRandcytokineinflammationproduction.
Conclusions: These results suggest that deregulation of miRNAs may be
andAllergyandImmunology,DepartmentofInternalMedicine,University
ofSouthFlorida,Tampa,FL;2DivisionofAllergy&Immunology,University involved in the pathogenesis of asthma and miR-150 may suppress in-
of South Florida and James A. Haley Veterans’ Hospital, Tampa, FL; flammationinasthmabyinhibitingcytokineproductionbydownregulating
3Nanomedicine Research Center and Divison of Translational Medicine, critical genes such as Akt, Elk1 and Cbl1. miR-150 may be an attractive
candidateforasthmagenetherapy.
Departments of Molecular Medicine and Internal Medicine, University of
SouthFloridaandJamesVHaleyVeterans’Hospital,Tampa,FL;4Nano-
medicineResearchCenterandDivisionsofTranslationalMedicine,Allergy
and Immunology, Departments of Internal Medicine and Molecular Medi-
cine, University of South Florida and James V Haley Veterans’ Hospital, 27
Tampa,FL. SerineProteaseInhibitorAttenuatesOvaInducedInflammation
Background:Weknowthataheterogeneousgroupofmyeloidcellstermed inMouseModelofAllergicAirwayDisease
myeloid derived suppressor cells (MDSC) accumulate in almost all Sanjay Sawc, MSc, Sagar Kalec, MSc, Bhanu Pratap Singh, PhD, and
pathologicalconditions,whichelicitaninflammatorysignal.Theexactrole Naveen Arora, PhD. Allergy and Immunology, Institute of Genomics and
playedbythesecellsinasthmaisnotknown.Inthisstudyweinvestigatedthe IntegrativeBiology,Delhi,India.
functionandroleofthesecellsinasthma. Background:Serineproteasespromoteinflammationandtissueremodeling
Methods:AccumulationofMDSCandothersubsetsofmyeloidcellswere by activating proteinase-activated receptors, urokinase, metalloproteinases
analyzed from peripheral blood mononuclear cells from patients with non- and angiotensin. In the present study, AEBSF (4-(2-Aminoethyl) benzene-
severeasthma(FEV).60)andsevereasthma(FEV ,60)bymulticolor- sulfonylfluoride)aserineproteaseinhibitor,wasevaluatedforprophylactic
1 1
flowcytometry and compared to healthy controls. Allergic mouse models andtherapeutictreatmentinmousemodelofairwayallergy.
wereusedtodeterminetheroleofmicroRNA-142(miR-142)inregulation Methods: BALB/c mice were sensitized by i.p route on 0 and 14 day and
andexpansionofMDSC. challengedwithOVA(25,26and27day)byi.n.route.Miceweretreatedi.n.with
Results:ThereisasignificantincreaseintheproportionofMDSCinsevere AEBSF,1hourbefore/afterchallengeandsacrificedonday29tocollectBALF,
versus non-severe asthmatics and controls, corresponding to a decrease in blood and lungs. OVA specific immunoglobulins were measured in serum.
myeloid dendritic cells. Allergic mice had significant increased levels of Proteolytic activity, total cell/eosinophil count, eosinophil peroxidase activity
MDSCexpansionwhichwereassociatedwithincreasedlevelsof IL-6and (EPO),IL-4,IL-5,IL-10,cysteinylleukotrienesand8-isoprostane(oxidativestress
downregulation of miR-142. miR-142 overexpression induced MDSC marker)weredeterminedinBALF.Haematoxylinandeosinstainedlungsections
differentiation. wereexaminedforcellularinfiltrationandairwayinflammation.
Conclusions:AnaccumulationofMDSCisassociatedwithsevereasthmain Results:MiceexposedtoOVAandtreatedwithPBSshowedsignificantly
humansandmice.Inanallergicmousemodel,IL-6levelsincrease.miR-142 high levels of IgE, IgG1 and IgG2a as compared to sham mice. Both
may play an important role in regulation and differentiation of MDSC, prophylactic and symptomatic AEBSF treatment reduced serum IgE and
leadingtoalteredimmunity. IgG1significantly(P#0.05)thancontrol,howevertherewaslittleincrement
(cid:1)2012WorldAllergyOrganization S9
Abstracts WAOJournal(cid:2)February2012
in IgG2a level. AEBSF could effectively reduce the proteolytic activity in amajorsourceofallergenbutwasrareinRomania.Theaimistoevaluate
BALF.IL-4andIL-5decreasedsignificantly(P#0.05)afterAEBSFtreat- the symptoms and associated factors in patients with allergic rhinitis to
mentwhileasignificant(P#0.05)increasewasobservedinIL-10inBALF. ragweedpolleninthenorthwestregionofRomania.
Airwayinflammationreducedsignificantlyasrevealedbylunghistopathol- Methods: 74 patients (pts) (mean age 27.97 6 13.85 years) with allergic
ogy, EPO activity and cysteinyl leukotrienes in BALF after treatment. rhinitis to ragweed pollen were included in the study. The patients were
AEBSFalsosuppressedoxidativestressintermsof8-isoprostaneinBALF. clinicallyevaluatedregardingtheseverityofthesymptomsonascalefrom
Amongthetreatmentdoses,10and50mgofAEBSFweremosteffectivein 0 to 3 and their duration. A total score over 6 indicates a moderate/severe
reducingmajorityoftheinflammatoryparameters. formofrhinitis.Weevaluatedtheassociationwithotherallergicmanifesta-
Conclusions: Prophylactic and therapeutic treatment of AEBSF attenuates tions(asthmaandurticaria).Allthepatientshadskinprickteststoinhalant
theairwayinflammationinmousemodelofairwayallergyandhavepotential allergens. The obtained data were statistically analyzed using Anova, Chi-
forthetreatmentofinflammatoryallergicdiseases. squareandFischertests,withasignificanceofP,0.05.
Results: 50.94% of the pts were female. 58.1% of them presented mild
allergic rhinitis, while 41.9% moderate severe forms. 27% of the pts were
monosensitisedtoragweedpollenand73%oftheptswerepolisensitised.The
28 patientsmonosensitisedtoragweedhadmoderate/severeformsofrhinitis(14
PotentialRoleofScavengerReceptorsinHumanMastCell vs86%,P¼0.004)comparedwithpolisensitisedgroup.Thesymptomsscore
CytokineResponsetoOxidizedldl washigherinptswithmonosensitisationcomparedwithpolisensitisationpts
RitaFaigerc,MSc,1JaniLappalainenc,MSc,2WolfgangJ.Schneider,PhD,3 (7.05vs5.28,P¼0.02).Inmonosensitizedgrouptheocularszmptomswere
PetriKovanen,MD,PhD,2andFrancescaLevi-Schaffer,PhD4.1Department morefrequentlypresent(65vs18%,P¼0.02)andweremoresevere(0.65vs
of Pharmacology, The Hebrew University of Jerusalem, Jerusalem, Israel; 0.33,P¼0.01).Thenumberofptswithassociationofallergicrhinitisand
2Wihuri Research Institute, Helsinki, Finland; 3Department of Molecular asthmawas higherin thepolisensitisedgroupcomparedto the monosensi-
Genetics,UniversityofVienna,Vienna,Austria;4DepartmentofPharmacol- tisedone(44.4vs11.11%,P¼0.029).Theintervalbetweentheonsetofthe
ogy,InstituteforDrugResearch,SchoolofPharmacy,FacultyofMedicine, symptomsanddiagnosisofrhinitisishigherinpolisensitisedptsandsignif-
TheHebrewUniversityofJerusalem,Jerusalem,Israel. icantlyincreasedinptswithasthma.Thereisnocorrelationbetweenenvi-
Background:Humanatheroscleroticlesionscontainmastcellsandoxidatively ronment(rural–urban),age,sex,familyandpersonalallergichistoryandthe
modifiedlow-densitylipoproteinparticles(oxLDL).Scavengerreceptorsarecell typeofsensitisationandseverityofthesymptoms.
surfacereceptorsthatbindandinternalizeoxLDL,andtheyplayanimportant Conclusions: Ragweed produces intense allergogen pollen and determines
role in macrophage foam cell development, a key event in the initiation and severeformsofallergicrhinitisandalsothepresenceofocularsymptoms.
developmentofatheroscleroticlesions.Thepurposeofthestudywastoanalyze Polisensitisationincreasestheriskofassociatedasthmaandalsoincreasesthe
expressionofthemostcommonscavengerreceptorsinmastcells,anddetermine intervalbetweeentheonsetofthesymptomsanddiagnosis.
whetheroxLDLparticlescaninducethemtosecretepro-inflammatorycytokines
thatarepotentiallycapableofinducingandamplifyingatherogenicprocesses.
Methods: Mast cells were differentiated from human cord blood-derived
CD341progenitorcellsinvitro(CBMC),andtheirexpressionofscavenger 30
receptorswasanalyzedbyconventionalRT-PCR,flowcytometryandWest- RagweedAllergy–WhatRoleDoesItPlayinBavaria?
ern blot techniques. Fluorescently-labeled oxLDL was used to investigate MareikeMcIntyre,1,2TeresaJaeger,1,2JohannesHuss-Marp,1,2MichaelHauser,3
LDLinternalizationbymastcells.Secretionofpro-inflammatorycytokines Fatima Ferreira,3 Markus Ollert,1 Johannes Ring,1 Carsten Schmidt-Weber,2
intotheincubationmediumanddegranulationofthemastcellsinresponseto Heidrun Behrendt,2 and Ulf Darsow,1,2. 1Department of Dermatology and
oxLDLwereassayedbyELISAandacolorimetric-enzymatictestforbeta- AllergyBiederstein,TechnischeUniversitaetMuenchen,Munich,Germany;
hexosaminidase,respectively. 2Center of Allergy and Environment (ZAUM), Helmholtz Center Munich/
Results:CBMCexpressedmRNAandproteinforLOX-1,SR-AIandCD68, Technische Universitaet Muenchen, Munich, Germany; 3Department of
butnotforCD36,andtheexpressionofLOX-1andSR-AIwasupregulatedby MolecularBiology,UniversityofSalzburg,Salzburg,Austria.
incubationofthecellswithoxLDL.CBMCinternalizedoxLDLmoreefficiently Background:Ragweed(Ambrosiaartemisiifolia),isincreasinglyspreading
thannativeLDL,whilesimultaneousneutralizationofCD68,SR-AIandLOX-1 in Southern Germany and Central Europe. Little is yet known about the
with monoclonal antibodiesresultedinreducedoxLDL uptake.Moreover, in sensitizationandallergyratesinBavaria.
responsetooxLDL,CBMCshowedincreasedreleaseofb-hexosaminidase,and Methods:In2008to2010patientsfromaBavarianuniversityallergyunit
adose-dependentsecretionofthepro-inflammatorycytokinesIL-8andMCP-1. wereenrolledintothestudy.Thepatient’shistorywasrecordedbyastand-
Conclusion: Our results reveal that cultured human mast cells express ardisedquestionnaireconcerningallergies.Sensitizationratesweremeasured
scavengerreceptorsthatareupregulatedbyoxLDL.Inatheroscleroticlesions, by skin prick test (SPT) for seasonal aeroallergens including ragweed.
oxLDLmayactivateMCtosecretepro-inflammatorycytokines,andsothey Patients sensitized to ragweed were further characterized by measuring
causemastcellstoactasacellularlinkbetweenoxLDLandtheinflammatory specific serum immunoglobulin E (IgE) for ragweed specific allergens
responseinatherosclerosis. (byImmunoCAPandELISA).Todeterminetheclinicalrelevancechallenge
tests(nasal/conjunctival)withragweedwereperformed.
Results:1022patientswereenrolledinthestudy(665female,357male).289
ALLERGIC RHINITIS
patients were sensitized to ragweed (SPT positive). In ragweed sensitized
patients the sensitization rate to mugwort was 61.8% whilst in patients not
sensitizedtoragweeditwas7.4%.Thesensitizationtobirchwas78.1%resp.
29 36.4%.In120ragweedsensitizedpatientschallengetestswithragweedextract
AllergicRhinitistoRagweedPollen wereperformed(nasaln¼110;conjunctivaln¼60)withpositiveresultsin29
IoanaCorinaBocsan,1IoanaAdrianaBujor,MD,2CristinaBarbinta,MD,3 (26%)resp.12(20%)patients.In232ragweedsensitizedpatientsspecificIgE
and Diana Deleanu, MD, PhD2. 1Clinical Pharmacology; 2Immunology- tonArtv1wasobservedsignificantlymorefrequentlythantonAmba1.
Allergology,UniversityofMedicineandPharmacy,Cluj-Napoca,Romania; Conclusions:Theresultsofthis3-yearstudyshowthatinaBavarianallergy
3Allergology,AlmedoClinic,ClujNapoca,Romania. unit sensitization to ragweed is frequent. Often ragweed-sensitized patients
Background:Theprevalenceofpollenallergyisestimatedaround40%in havesensitivitiestomultipleseasonalaeroallergens.Thereisacoexistenceof
general population. Ragweed (ambrosia artemisiifolia) pollen represents ragweed and mugwort specific allergens. One fourth of the challenged
S10 (cid:1)2012WorldAllergyOrganization