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20JUNE2010NatureMatreialsNanogelantigenicprotein-deliverysystemforadjuvant-freeintranasalvaccinesNanotechnologyisaninnovativemethodoffreelycontrollingnanometre-sizedmaterials1.Recentoutbreaksofmucosalinfectiousdiseaseshaveincreasedthedemandsfordevel-opmentofmucosalvaccinesbecausetheyinducebothsys-temicandmucosalantigen-specificimmuneresponses2.Herewedevelopedanintranasalvaccine-deliverysystemwithananometre-sizedhydrogel(‘nanogel’)consistingofacationictypeofcholesteryl-group-bearingpullulan(cCHP).Anon-toxicsubunitfragmentofClostridiumbotulinumtype-AneurotoxinBoHc/AadministeredintranasallywithcCHPnanogel(cCHP–BoHc/A)continuouslyadheredtothenasalepitheliumandwaseffectivelytakenupbymucosaldendriticcellsafteritsreleasefromthecCHPnanogel.Vigorousbotulinum-neurotoxin-A-neutralizingserumIgGandsecretoryIgAantibodyresponseswereinducedwithoutco-administrationofmucosaladju-vant.Importantly,intranasallyadministeredcCHP–BoHc/Adidnotaccumulateintheolfactorybulbsorbrain.Moreover,intranasallyimmunizedtetanustoxoidwithcCHPnanogelin-ducedstrongtetanus-toxoid-specificsystemicandmucosalim-muneresponses.TheseresultsindicatethatcCHPnanogelcanbeusedasauniversalprotein-basedantigen-deliveryvehicleforadjuvant-freeintranasalvaccination.题目：一种由纳米凝胶构成的新型抗原蛋白疫苗传递系统---通过鼻腔给药而无需要其他辅佐剂摘要：作为一种新手段，纳米技术可以生产纳米级的新材料。最近爆发的粘膜传染疾病会引起全身性以及粘膜特异的免疫应答。因此，粘膜抗体对人类的健康是至关重要的。这里，我们研究了一种新型的疫苗传递系统，这是一种由带正离子的胆甾醇基和支链淀粉构成的纳米级凝胶(cCHP)。一种无毒的蛋白片段---肉毒杆菌神经毒素A抗原疫苗Hc(BoHc/A)与cCHP纳米凝胶一起通过鼻腔给药方式粘附到鼻腔粘膜上皮细胞，BoHc/A从cCHP纳米凝胶中释放后有效地进入了粘膜的树突状细胞并且引起强烈的IgG、IgA免疫应答。这种免疫反应并不需要其他的辅助剂。更重要的是，鼻腔给药的cCHP–BoHc/A并不会在嗅球或者大脑中积累。此外，用这种纳米凝胶载体制作的破伤风疫苗也获得了特异的全身性免疫应答反应。这些结果显示cCHP可以作为一种普遍的蛋白抗原传递载体，用作生产无其他辅助剂的鼻腔给药疫苗。Beginningin2003,anenormousresearchinitiative—GrandChallengesinGlobalHealth—hasbeenorganizedworldwidewiththesupportoftheBillandMelindaGatesFoundationandtheUSNationalInstitutesofHealth.Itsaimistoovercometheglobalinfectiousdiseaseproblemsaffectinghumanhealthtoday.Thedevelopmentofanew-generationneedle-freemucosalvaccinehasbeenproposedasoneoftheinitiative’smostimportantgoals,becauseitcanelicitantigen-specificsystemichumoralandcellularimmuneresponsesandsimultaneouslyinducemucosalimmunity,especiallyintheaero-digestiveandreproductivetracts2–4.FluMist,whichiscomposedofcold-adaptedtrivalentliveinfluenzaviruses,isawell-knownexampleasthefirstadvancedintranasalvaccinetobeusedinUSpublichealth,in2003(ref.5).Sincethen,tremendouseffortshavebeenmadetofurtherdevelopintranasalvaccinetechnology.Subunitintranasalvaccinationisexpectedtobethesafeststrategy,becauseitshouldhavealowriskofcausingunfavourableandundesiredbiologicalreactions.However,intranasaladministrationofasubunitantigenaloneisgenerallyinsufficientforinductionofantigen-specificimmuneresponses.Asaresult,anadjuvantsuchasabacterialtoxingenerallyneedstobeadded,butthesetoxinsarepoorlytoleratedbyhumans.从2003年开始，一个庞大的科学计划-人类全球健康的挑战-开始实施，这项计划是由比尔盖茨基金会以及美国国家卫生研究院资助，主要目的是克服全球传染性疾病对人类健康的影响。生产粘膜吸收类的疫苗是其中重要的一个目标，因为这种疫苗可以引起特异的全身性细胞免疫应答以及获得粘膜免疫能力。尤其是在呼吸道和生殖系统。在美国，流感疫苗是第一个被成功研制的鼻内给药疫苗（2003）。从那以后，科学家们努力研发其他的鼻内给药疫苗。这种抗原片段鼻内给药疫苗不易引起不适和不良反应，因此被人们认为是安全的。然而这种疫苗存在一个很大的问题,在鼻内给药后很难引起特异的免疫应答。所以，一些辅助剂（细菌毒素等）需要添加到药物中，但是这些毒素却通常不被人类所耐受。Choleratoxinandheat-labileenterotoxinhavebeenextensivelyusedaspotentmucosaladjuvantsinexperimentalanimalstudiesbecauseoftheirmultipleimmune-potentiatingfunctions:theyactivateimmunocompetentcells,includingdendriticcellsandBcells,andthusinduceantigen-specificmucosalimmunity.However,ahumanclinicaltrialcarriedoutinSwitzerlandfrom2000to2001todevelopanintranasalinfluenzavaccinewithinactivatedinfluenzaviruscombinedwithasmallamountofheat-labileenterotoxinwaswithdrawnbecausetheco-administeredheat-labileenterotoxinwassuspectedofcausingBell’spalsy,ararecondition,invaccinatedsubjects.Inaddition,aseparatestudyinmicedemonstratedthatthetoxin-basedadjuvantmigratedinto,andaccumulatedin,theolfactorytissues.Asaresultofthesesafetyissues,thedevelopmentofintranasalvaccinesemployingtheco-administrationoftoxin-basedadjuvantshasrapidlydeclined.Furtherscientificandtechnologicalinnovationsthatwillhelpthedevelopmentofsafebuteffectiveadjuvant-freeintranasalvaccinesare,therefore,ofhighpriorityinglobalhealth.Applicationofbiomaterials,suchaspolymernanoparticlesandliposomes,hasagreatpotentialinvaccinedevelopmentandimmunotherapy.Inparticular,nanometre-sized(100nm)polymerhydrogels(nanogels)haveattractedgrowinginterestasnanocarriers,especiallyindrug-deliverysystems.霍乱毒素和不耐热肠毒素具有多重的免疫加强功能，他们在动物实验中可以作为粘膜吸收类抗原疫苗的辅佐剂。他们可以激活免疫活性细胞，包括树突细胞和B细胞，进而引起特异的粘膜免疫反应。然而，2000到2001年在瑞士的一个临床试验中，一种由不耐热肠毒素作为辅佐剂的鼻内给药流感疫苗失败了。因为这种疫苗接种后被怀疑会导致一种小概率的贝尔氏麻痹。另外一个独立的研究显示，这种辅佐剂会转移并在嗅觉系统中累计。因为这些安全因素，使用不耐热肠毒素作为辅佐剂的鼻内给药疫苗已经停止研究。发展的科学技术革新帮助我们研究出安全的不需要辅佐剂的鼻内给药疫苗为人类的健康做贡献。新型生物材料，诸如纳米聚合体、脂质体，为疫苗的研究和免疫治疗带来了巨大的发展空间。尤其是纳米级别的纳米水凝胶在作为药物传递系统载体方面有很好的前景。Wehavedevelopedanewmethodofcreatingaseriesoffunctionalnanogelsthroughself-assemblyofassociatingpolymers.Oneofthesepolymers,thecholesteryl-group-bearingpullulan(CHP)formsphysicallycrosslinkednanogelsbyself-assemblyinwater(Fig.1aandSupplementary.TheCHPnanogelstrapvariousprotei