dietaryricebrandemonstratesgreaterfeasibilityandmaybemorepracticalforp - PDF document

dietaryricebrandemonstratesgreaterfeasibilityandmaybemorepracticalforpublichealthinterventioninthedevel-opinganddevelopedworldwhencomparedwithextractpreparations.Giventhestrongpotentialfordietaryinclusionofwholericebranasafunctionalfoodingredientforchronicdiseasecontrolandprevention,studiesfocusedontheeffectsofdietaryricebranintakeonthegutmucosalim-muneresponseandthemicrobiotaarewarranted.Wein-vestigatedwhethera10%ricebranintakewhencomparedwithacontroldietledtochangesincellularandhumoralimmuneresponsesaswellasalterationsintheconcentrationofnativegutMATERIALSANDMETHODSAnimalsandfeedingscheduleSpeciÞcpathogen-free4Ð6-week-oldfemaleICRmicewerepurchasedfromHarlanLaboratories(Indianapolis,IN,USA).SpeciÞcpathogen-freemicehaveanormalcomposi-tionofintestinalbacteriaandaredevoidofknownmousepathogens.Allmicewereprovidedwateradlibitumandfedamaintenancediet(AIN-93M;HarlanTeklad,Madison,WI,USA)for1weekpriortorandomization.Micewereplacedoneitherthecontroldiet(AIN-93M)orthe10%ricebrandietfor28days.TheInstitutionalAnimalCareandUseCom-mitteeatColoradoStateUniversityapprovedallprotocolsinvolvingtheanimalexperimentsdescribedinthisstudy.DietcompositionTheNeptunericevarietywaschosenbasedonitsavail-abilityforricegrownintheUnitedStates.Control(AIN-93M)andricebranÐcontainingdietswereformulatedtomatchmacronutrientsacrossdietgroups,anddifferencesintotalstarchandÞbercontentswerebalancedusingpuriÞeddietcomponents.ThecompositionofricebranÐcontainingdietwascalculatedbasedonpublishedreportsdemonstratedchronicdisease-ÞghtingactivityofricebranandwerestoredatÐ20Cuntilfedtothemice.Dietfor-mulationsareshowninTable1.SamplingproceduresMicewerefedtheexperimentalricebrandietfor28days.Fecalandserumsampleswerecollectedfromallmiceonday0,4,7,14,18,21,25,and28followingcommencementofexperimentaldiets.Micewerebledfromthetailvein(50ÐLpermouse),andserumsampleswereobtainedusingserumseparatortubes(BDBiosciences,SanDiego,CA,USA).Fecalextractsweremadebyusingmethodspreviouslydescribed.QuantiÞcationoftotalIgAandIgAantibodyTotalIgAantibodytiterswereassessedinmousefecesandserumbyenzyme-linkedimmunosorbentassay(ELISA).Nunc-Immunoplates(ThermoScientiÞc,Waltham,MA,USA)werecoatedovernightat4CwithpuriÞedratanti-mouseIgA(C10-3;BD)ataconcentrationof5g/mLincarbonate-bicarbonatebuffer(pH9.6).NonspeciÞcproteinbindingsiteswereblockedwithphosphate-bufferedsaline(PBS)containing3%bovineserumalbumin(Sigma,St.Louis,MO,USA).Dilutions(1%bovineserumalbumin)ofthesamplesandstandardwereappliedtotheplate(50Lperwell)andincubatedfor1.5hoursatroomtemperature.Plateswereincubatedwithbiotinratanti-mouseIgA(C10-1;BD)atadilutionof1:1,000for1houratroomtemperature.Next,plateswereincubatedwithperoxidase-conjugatedstreptavi-din(JacksonImmunoResearch,WestGrove,PA,USA)ataconcentrationof1:1,500for20minutesatroomtemperature.Platesweredevelopedwith3,3,5,5-tetramethylbenzidine(Sigma).Thereactionwasstoppedusing1HCl,andtheopticaldensitieswerereadat450nm.PuriÞedmouseIgA(eBioscience,SanDiego,CA,USA)wasusedtocreatethestandardcurvefortheELISA.IgAELISAusedasimilarprotocolasthetotalIgAELISA.Thespp.usedtocoattheplatewereisolatedfromfecalsamplescollectedonday18followingcommencementofdietaryricebran.Thelacto-bacilliwasexpandedinMRSbrothpriortoundergoingheat-killingasdescribedpreviously.ELISAplateswerecoatedovernightat4Cwith10colony-formingunits/mLheat-killedlactobacilliincarbonate-bicarbonatebuffer.Non-speciÞcbindingsiteswereblockedwithPBScontaining5%nonfatdriedmilk.ApositivecontrolsamplewasobtainedbyvaccinatingintraperitoneallyanICRmousewiththeheat-killedlactobacilliandcollectingserumafter14days.TissueharvestandimmunecellpreparationAfterthe28-dayfeedingperiod,micewerekilledfol-lowinganesthetizationbyintraperitonealinjectionofketa-mine(100mg/kg)withxylazine(10mg/kg).ThemesentericlymphnodesandPeyerÕspatcheswerecollected,me-chanicallydisruptedthroughameshscreen,andtreatedwith CompositionofExperimentalDietsLevel(g/kg)ComponentControldiet10%ricebrandietCasein140.0140.0-Cysteine1.81.8Cornstarch465.7422.7Maltodextrin155.0155.0Sucrose100.0100.3Cornoil40.019.0Cellulose50.029.0MineralMix35.035.0VitaminMix10.010.0Cholinebitartrate2.52.5TBHQ,antioxidant0.0080.008RicebranÑ100.0Controldiet(AIN-93M)producedbyHarlan.10%ricebrandietproducedbyHarlanusingAIN-93MinadditiontoricebranfromNeptunericevariety,adjustedtomatchcontroldietintotalfat,protein,andcarbohydrates.-butylhydroquinone.470HENDERSONETAL. Cl.Next,a10-cmsectionoftheintestine(ileum)wascollected,andtheintestinallaminaproprialymphocyteswerepreparedaspreviouslydescribed.Theintestinaltis-suewasdisruptedthroughameshscreenpriortobeingrunthroughadensitygradient(Optiprep,AccurateChemical&ScientiÞcCorp.,Westbury,NY,USA).AllcellpreparationswereresuspendedincompleteRPMI1640medium(In-vitrogen,Carlsbad,CA,USA)containing10%fetalbovineserum(GeminiBio-Products,WestSacramento,CA,USA),-glutamine(Invitrogen),1nonessentialaminoac-ids(Invitrogen),0.075%sodiumbicarbonate(FisherSci-entiÞc,Pittsburgh,PA,USA),100U/mLpenicillin,andg/mLstreptomycin(Invitrogen).Cellswerekeptonicepriortoimmunostainingandanalysis.FlowcytometricanalysisDirectlyconjugatedantibodiesusedfortheseanalyseswerepurchasedfromeBioscienceorBDPharmingen(SanJose,CA,USA).Thefollowingantibodieswereusedinvariouscombinations:anti-CD4(GK1.5),anti-CD8b(H35-17.2),anti-CD5(53-7.3),anti-CD27(LG.7F9),anti-CD11c(N418),anti-CD11b(M1/70),anti-CD45R(cloneRA3-6B2),andanti-mouseIgA(C10-1).Immunostainingwascompletedasdescribedpreviously.AnalysiswasperformedwithaCyanADPßowcytometer(BeckmanCoulter,FortCollins,CO,USA),anddatawereanalyzedusingFlowJosoftware(TreeStar,Ashland,OR,USA).Onday0,4,7,14,18,21,25,and28,followingcom-mencementofexperimentaldiets,fecalsampleswerecol-lectedfromallmice.ApproximatelyÞveorsixfreshfecalpellets(0.1g)werecollectedpermouseandrehydratedin1mLofsterilePBSfor15minutes.Thesampleswerevortex-mixed,dilutedinPBS,platedonMRSagar(enrichmentagarforthegenusLactobacillus),andplacedina37Cincubatorcontaining5%COfor48hours.AllcoloniesgrownonMRSagarwereconÞrmedasLactobacillusspp.usingreal-timepolymerasechainreactionaspreviouslydescribed.StatisticalanalysesStatisticalanalysiswasperformedusingPrismversion5.0software(GraphPad,LaJolla,CA,USA).Two-tailednonparametric(MannÐWhitney)testswereperformedforcomparisonsbetweenthetwogroups.Arepeated-measures(mixedmodel)two-wayanalysisofvariancewithaBon-posthoctestwasperformedtocomparethetwogroupsovertime.DifferenceswereconsideredstatisticallysigniÞcantfor.05forallcomparisons.DietaryricebranintakeinduceslocalandsystemicIgAproductionTheeffectofdailyricebranintakeonthemucosalim-munesystemwasÞrstinvestigatedinmiceforchangesinserum,intestinal,andfecalIgA.Controland10%ricebrandietswerefedtomicefor28days,andthetotalIgAanti-bodylevelsweremeasuredthroughoutthistimeperiod.TheconcentrationoftotalIgAinthefecesofthericebranÐfedmice(Fig.1A)wassigniÞcantlyincreasedonday18andremainedhighthroughday21incomparisonwithmiceonthecontroldiet.Asimilardifferencewasdetectedsystem-icallyintheserum(Fig.1B)withthetotalIgAconcentrationpeakingatday18.WeassessedtheexpressionofIgAonthesurfaceofin-testinalBcellstoelucidatethemodulatingIgAresponsefollowingdietaryricebranintake.IncreasedexpressionofsurfaceIgAindicatesactivationoflocalBcellsaswellasan FIG.1.Effectofdietaryricebranonlocalandsystemictotalim-munoglobulinA(IgA)responses.AtotalIgAenzyme-linkedimmu-nosorbentassaywasperformedonserialdilutionsof(A)fecaland(B)serumsamplescollectedfor28days.Antibodyconcentrationsweredeterminedthroughcomparisonwithastandardcontrol.DataaremeanSEMvalues(12micepergroup).Resultswerepooledfromtwoindependentexperiments.SigniÞcantdifferences(***.001)weredeterminedbyarepeated-measures(mixedmodel)two-wayanalysisofvariancefollowedbyaBonferroniposthoctest.RICEBRANAIDSMUCOSALIGAANDLACTOBACILLI471 increasedpotentialtorespondtocommensalandpathogenicbacteriainaTcellÐindependentfashion.Micewereeu-thanizedafter14daysofricebranconsumption,andtheBcellswereanalyzedforsurfaceIgAexpression(Table2).AsigniÞcantdifferencewasobservedinthenumberofIgAmoleculesexpressedontheBcellsurfacebetweencontrolandricebranÐfedmice,determinedbymeanßuorescenceintensity.Theseresultsareconsistentwiththeideathatdietaryricebraninducesmucosalim-munityviasystemicandlocalIgAproductionaswellasincreasedactivationofIgABcellsfoundinthePeyerÕsIncreasedmucosalCD11cdendriticcellsinducedbyricebranInordertodeterminewhetherdietaryricebranintakeinßuencesantigen-presentingcellsasamechanismforIgAinduction,weexaminedcellularimmunephenotypespre-viouslyshowntoberequiredforIgAclassswitching.Following14daysofdaily10%ricebranintake,miceweresacriÞced,andthecellularresponseswereassessedinthelaminapropria,PeyerÕspatches,andmesentericlymphnodes(Table3).Thepercentagesofmyeloiddendriticcells)inboththelaminapropriaandthemesentericlymphnodesweresigniÞcantlyincreasedinmicefedthe10%ricebrandietincomparisonwiththemiceoncontroldiet.NosigniÞcantdifferencesweredetectedinTorBcellpopulations(Table3).ThesedatasuggestedthatricebranÐmediatedinductionofdendriticcellrecruitmentintothemucosaltissuesresultedinincreasedantigenpre-sentationandsubsequentIgAproduction.DietaryricebranincreasedGiventheassociationsofprobioticbacteriawiththemodulationofthemucosalimmunesystem,wenextde-terminedthetitersofthenativegutspp.inthericebranÐfedmicecomparedwiththecontrolgroup.waschosenforitsaerobicgrowthconditionsaswellasitsabilitytoprotectagainstavarietyofinfectiousFortheseexperiments,weprocessedandplatedfreshfecalpelletsforthepresenceofspp.overa28-dayperiod.Ondays11,18,21,and25thenumbersofspp.inthefecesweresigniÞcantlyhigherfromthericebranÐfedmiceincomparisonwithamorecyclicalpatternseeninthemicefedcontroldiet(Fig.2).Thenearly500%increaseinnumbersof(incolony-formingunitspergramoffeces)becameappar-entonday11andremainedsigniÞcantlyhighfor14days.Becauseoftheabilityofmultiplespeciestoinduceprotectionagainstintestinalpathogens,polymerasechainreactionwasperformedtoconÞrmagreaterthan900%increaseinDNAlevelsfromthegenus IncreasedExpressionofSurfaceImmunoglobulinCellsinPeyer’sPatchesDaysofRiceBranDietConsumptionLaminapropriaPeyerÕspatchesMesentericlymphnodesCellpopulationControl10%RBControl10%RBControl10%RBcells(%)1.40.31.60.31.00.31.10.30.60.10.7IgA(MFI)onB220Bcells193361945198.723.1166.719.7*81.32488.6DataaremeanSEMvalues,pooledfromtwoindependentexperiments(.05asdeterminedbycomparingcontroldietversus10%ricebran(RB)dietineachcellpopulationusinganonparametricMannÐWhitneyMFI,meanßuorescenceintensity. InuenceofDietaryRiceBranonCellularPopulationsintheIntestinalImmuneCompartmentsFollowingDaysofDietConsumptionLaminapropriaPeyerÕspatchesMesentericlymphnodesCellpopulation(%)Control10%RBControl10%RBControl10%RBTcells0.89.61.020.31.519.71.966.81.767.63.318.43.36.30.84.50.417.10.616.3Bcells5.931.33.166.13.666.44.110.83.620.40.11.30.13.30.73.80.87.41.58.70.21.00.30.20.040.20.090.30.050.3DCs6.80.911.10.9**1.10.11.20.10.60.10.7DataaremeanSEMvalues,pooledfromtwoindependentexperiments(.05,**.01asdeterminedbycomparingcontroldietversus10%RBdietineachcellpopulationusinganonparametricMannÐWhitneyDC,dendriticcell.472HENDERSONETAL. betweenday0andday14inthemicefedthericebrandiet(datanotshown).Boththecontrolandricebrandietsdidnotcontainanylactobacilli.Next,wedeter-minediftheincreasedIgAtiterswerespeciÞc.AIgAELISAwasperformedandrevealedlowtoundetectable-speciÞcIgA(datanotshown).ApositivecontrolwasinplacetoensurethereliabilityandsensitivityoftheELISAasdescribedinMaterialsandMethods.Therefore,theseresultsindicateapotentialrolefordietaryricebraninmodulatingtheintestinalmicrobiotathroughelicitingin-creasedconcentrationsofbeneÞcialbacteria.Theuniquehealth-promotingpropertiesandchemicalcompositionofricebranmakeitapromisingcandidatefordietarysupplementation,fornutritionaltherapy,andforpreventionofchronicdisease.1,2,4Ð9,28,44Ð47InthisstudywefoundtheeffectsofwholedietaryricebranintakeonthemucosalimmunesystemtobetheinductionofIgA(Fig.1)andtheenhancementoftheinnateimmuneresponse(Table3).Inaddition,theabilityofdietaryricebrantopromotein-creasedintestinalcolonizationofnative(Fig.2)highlightsanovelroleforricebranprebioticcomponentstoinßuencetheintestinalmicrobiota.Understandingtheeffectofdietaryricebranontheinnateimmunesystemandantigenpresentationiscrucialtoelu-cidatingthemechanismsbywhichricebranmayinduceprotectiveresponsesatmucosalsurfaces.Theincreasedpercentagesofmyeloiddendriticcellsinboththelaminapropriaandthemesentericlymphnodesfollowingricebranconsumptionspeaktothetargetedaffectofricebranonthekeycellsinvolvedinshapinganimmuneresponse.In-testinaldendriticcellshavetheuniqueabilitytoinduceIgAproductionfromBcellsthroughsecretionofretinoicacid,interleukin-5,andinterleukin-6.Therefore,theenhanceddendriticcellpopulationcanhelpintheinitiationofanadaptiveimmuneresponsethroughtheactivationofBcellsandsubsequentIgAclass-switching.Also,theincreasedpresenceofdendriticcellsinthelaminapropriaenhancesthemucosalinnateimmuneresponseatthesitewherepathogensinvadeandrevealsapotentialprotectivemech-anisminducedbydietaryricebran.EvidenceforthemodulationofthemucosalIgAresponseassociatedwiththeconsumptionofdietarycomponentshasbeenlimitedtofructooligosaccharidesandpectin.ConsistentwiththeseÞndings,thisstudydemonstratedtheabilityofdietaryricebrantoinduceincreasedIgAcon-centrationssystemicallyandlocally.Variousbioactiveandprebioticcomponentspresentinricebranarehypothesizedtobeinvolvedintheenhancementofimmunity.Thesephytochemicalsinclude,butarenotlimitedto,polyphenols,andfattyacids,aswellassomeessentialaminoacidsandmicronutrients.Understandingthemech-anismofIgAinductionisofconsiderableimportancewhenevaluatingthedietarycapacityofricebrantoelicitpro-tectionagainstentericinfections.CurrentevidencesuggeststhatthemajorityofIgAmoleculesinthegutareinducedbycommensalbacteriaandthatthereisaroleforthistransientpopulationtoinduceaspeciÞcIgAresponse.ArecentstudyperformedbyHapfelmeieretalconÞrmstheroleofcommensalbacteria,butalsodescribesthespeciÞcIgAre-sponsetobelessrobustandhaveasloweronset,consistentwithourÞndings.AnotherrelevantÞndingbyMacphersonetalshowedinductionofcommensal-speciÞcIgAanti-bodiestobemostlyTcellindependentbutdependentlar-gelyontheB1peritonealBcells,apotentialanglethatshouldbeevaluatedinricebranÐfedmice.Basedontheemergingevidenceforcommensalin-volvementinshapingthemucosalIgApopulation,thebeneÞcialeffectofdietaryricebranonthenativegutspp.wasshowntobeapotentialmechanismofimmunemodulation(Fig.2).Theantibody-enhancingabilityofwasdescribedpreviouslyinastudyshowingincreasedproductionofrotavirus-speciÞcanti-bodieswhenfermentedmilkwasadministeredduringtheacutephaseofarotavirusinfection.Thelow-speciÞcIgAantibodiesinourmodelmayspeaktotheuniqueabilityofricebrantoenhanceantibodyresponsesspeciÞcforotherbacteria,oritmayreßectthefactthatdietaryricebrancontainsgrowthsubstratesforresidentbacteria.Asaresult,thespeciÞcityoftheincreasedIgAwouldbeheterogeneous,makingthetitertoolowfordetectionbyELISA.WehypothesizethatricebranÐinducedmodulationofmultiplecommensalbacteriaresultedinincreasedluminalIgAconcentrationsintheintestine.Futureresearchdirectionsshouldincludeeluci-datingtheeffectofdietaryricebranonthegastrointestinalmicrobiotausing454pyrosequencingorotherhigh-throughputmicrobiomeanalyticalapproaches.Theabilityofdietaryricebrantopromotethegrowthofnativegutspp.andenhancemucosalimmunecellpopulationsoffersnumeroushealth-promotinganddisease-Þghtingpossibilities.Forexample,thepotentialuse FIG.2.EffectofdietaryRBonfecaltitersof.Freshfecalpelletswerecollected,processed,andplatedonMRSagarevery3Ð4daysinordertodeterminetheLogtiterpergramoffeces.DataaremeanSEMvalues(10micepergroup).Re-sultswerepooledfromtwoindependentexperiments.SigniÞcantdifferences(*.05,***.001)weredeterminedbyarepeated-measures(mixedmodel)two-wayanalysisofvariancefollowedbyaposthocRICEBRANAIDSMUCOSALIGAANDLACTOBACILLI473 ofricebranasadietaryvaccineadjuvantishighlightedbytherecentapplicationofLactobacillusfermentuminanin-tramuscularinßuenzavaccine.Also,thebeneÞcialeffectsofricebranoninßammatorydiseasessuchasinßammatoryboweldiseaseandtype1diabetesemphasizeaun-iqueresearchavenuetostudytheimmune-enhancingeffectsofdietaryricebranonthesenutritionallyrelevantdiseases.Insummary,theabilityofwholedietaryricebrantomod-ulatetheimmunesystemaswellaspromotethegrowthofnativegutspp.holdsgreatpromiseinpro-tectionagainstentericpathogensandmodulationofchronicinßammatorydiseases.WewouldliketothankDr.AnnaMcClungfromtheU.S.DepartmentofAgriculturericeresearchunitforprovidingricebranfromthesingleNeptunevariety.ThisworkwassupportedbyaGrandExplorationsinGlobalHealthgrant(OPP1015267)fromtheBillandMelindaGatesFoundationandtheShipleyFoundationandagrantfromtheNationalInstitutesofHealth(R03CA150070).AUTHORDISCLOSURESTATEMENTTheauthorsdisclosenoconßictsofinterest.1.HudsonEA,DinhPA,KokubunT,SimmondsMSJ,GescherA:Characterizationofpotentiallychemopreventivephenolsinex-tractsofbrownricethatinhibitthegrowthofhumanbreastandcoloncancercells.CancerEpidemiolBiomarkersPrev2.RyanEP:Bioactivefoodcomponentsandhealthpropertiesofricebran.JAmVetMedAssoc3.RyanEP,HeubergerAL,WeirTL,BarnettB,BroecklingCD,PrenniJE:RicebranfermentedwithSaccharomycesboulardiigeneratesnovelmetaboliteproÞleswithbioactivity.JAgricFoodChem4.CaiH,Al-FayezM,TunstallRG,etal.:ThericebranconstituenttricinpotentlyinhibitscyclooxygenaseenzymesandinterfereswithintestinalcarcinogenesisinApcMinmice.MolCancerTher5.NorazalinaS,NorhaizanME,HairuszahI,NorashareenaMS:AnticarcinogenicefÞcacyofphyticacidextractedfromricebranonazoxymethane-inducedcoloncarcinogenesisinrats.ToxicolPathol6.ChouTW,MaCY,ChengHH,ChenYY,LaiMH:Aricebranoildietimproveslipidabnormalitiesandsuppresshyper-insulinemicresponsesinratswithstreptozotocin/nicotinamide-inducedtype2diabetes.JClinBiochemNutr7.GerhardtAL,GalloNB:Full-fatricebranandoatbransimilarlyreducehypercholesterolemiainhumans.JNutr1998;128:865Ð869.8.HegstedM,WindhauserMM,MorrisSK,LesterSB:Stabilizedricebranandoatbranlowercholesterolinhumans.NutrRes9.KahlonTS,ChowFI,SayreRN,BetschartAA:Cholesterol-loweringinhamstersfedricebranatvariouslevels,defattedricebranandricebranoil.JNutr10.KataokaK,OgasaS,KuwaharaT,etal.:Inhibitoryeffectsoffermentedbrownriceoninductionofacutecolitisbydextransulfatesodiuminrats.DigDisSci11.KomiyamaY,AndohA,FujiwaraD,etal.:Newprebioticsfromricebranameliorateinßammationinmurinecolitismodelsthroughthemodulationofintestinalhomeostasisandthemucosalimmunesystem.ScandJGastroenterol12.GibsonGR,ProbertHM,LooJV,RastallRA,RoberfroidMB:Dietarymodulationofthehumancolonicmicrobiota:updatingtheconceptofprebiotics.NutrResRev13.IsolauriE,SutasY,KankaanpaP,ArvilommiH,SalminenS:Probiotics:effectsonimmunity.AmJClinNutr14.SandersME:Probiotics:considerationsforhumanhealth.15.CummingsJH,MacfarlaneGT:Roleofintestinalbacteriainnutrientmetabolism.ClinNutr16.ReillyKJ,RombeauJL:Metabolismandpotentialclinicalappli-cationsofshort-chainfattyacids.ClinNutr1993;12:S97ÐS105.17.vanHylckamaVliegJET,VeigaP,ZhangC,DerrienM,ZhaoL:ImpactofmicrobialtransformationoffoodonhealthÑfromfermentedfoodstofermentationinthegastro-intestinaltract.CurrOpinBiotechnol18.MakarovaK,SlesarevA,WolfY,etal.:Comparativegenomicsofthelacticacidbacteria.ProcNatlAcadSciUSA19.UlluwishewaD,AndersonRC,McNabbWC,MoughanPJ,WellsJM,RoyNC:Regulationoftightjunctionpermeabilitybyintestinalbacteriaanddietarycomponents.JNutr20.CorrSC,LiY,RiedelCU,OÕToolePW,HillC,GahanCGM:Bacteriocinproductionasamechanismfortheantiinfectiveac-tivityofLactobacillussalivariusProcNatlAcadSci21.ForsytheP,BienenstockJ:Immunomodulationbycommensalandprobioticbacteria.ImmunolInvest22.HeB,XuW,SantiniPA,etal.:IntestinalbacteriatriggerTcell-independentimmunoglobulinA2classswitchingbyinducingepithelial-cellsecretionofthecytokineAPRIL.23.MacphersonAJ,GattoD,SainsburyE,HarrimanGR,HengartnerH,ZinkernagelRM:AprimitiveTcell-independentmechanismofintestinalmucosalIgAresponsestocommensalbacteria.24.ToriiA,ToriiS,FujiwaraS,TanakaH,InagakiN,NagaiH:Lac-tobacillusacidophilusstrainL-92regulatestheproductionofTh1cytokineaswellasTh2cytokines.AllergolInt2007;56:293Ð301.25.PerdigonG,deMaciasME,AlvarezS,OliverG,deRuizHol-gadoAA:Effectofperorallyadministeredlactobacillionmac-rophageactivationinmice.InfectImmun26.PerdigonG,deMaciasME,AlvarezS,OliverG,deRuizHol-gadoAP:SystemicaugmentationoftheimmuneresponseinmicebyfeedingfermentedmilkswithLactobacilluscaseiLactobacillusacidophilus27.GaldeanoCM,PerdigonG:TheprobioticbacteriumLactobacilluscaseiinducesactivationofthegutmucosalimmunesystemthroughinnateimmunity.ClinVaccineImmunol2006;13:219Ð226.28.SierraS,Lara-VillosladaF,OlivaresM,JimenezJ,BozaJ,XausJ:Increasedimmuneresponseinmiceconsumingricebranoil.EurJNutr474HENDERSONETAL. 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ConsumptionofRiceBranIncreasesMucosalImmunoglobulinAConcentrationsandNumbersofIntestinalAngelaJ.Henderson,AjayKumar,BrittanyBarnett,StevenW.Dow,andElizabethP.RyanDepartmentsofMicrobiology,Immunology,andPathologyandClinicalSciencesandCenterforRhizosphereBiology,ColoradoStateUniversity,FortCollins,Colorado,USAABSTRACTGut-associatedlymphoidtissuemaintainsmucosalhomeostasisbycombatingpathogensandinducingastateof Manuscriptreceived15August2011.Revisionaccepted18November2011.Addresscorrespondenceto:ElizabethP.Ryan,AnimalCancerCenter,DepartmentofClinicalSciences,ColoradoStateUniversity,FortCollins,CO80523,E-mail:e.p.ryan@colostate.eduJOURNALOFMEDICINALFOODJMedFood15(5)2012,469–475MaryAnnLiebert,Inc.,andKoreanSocietyofFoodScienceandNutritionDOI:10.1089/jmf.2011.0213