人类免疫缺陷病毒（HIV）负性调控因子（Nef）是一个辅助蛋白，被认为是HIV相关的免疫和神经免疫发病机制的组成部分。在此，我们表明，Nef转染的小胶质细胞释放的Nef+exosome破坏顶部血脑屏障（BBB），而且只有Nef转染的小胶质细胞释放Nef入exosome。绿色荧光标记(gfp)的Nef转染的神经元和星形胶质细胞能够释放exosomes但不能释放exNef到细胞外间隙。Nef转染T细胞来源的exNef的顶部控制会减少跨内皮细胞电阻（TEER）并增加血脑屏障的通透性。小胶质细胞源性exNef应用在顶部/基底的血脑屏障能显着降低紧密连接蛋白，ZO-1，表明exNef介导神经病理的发病机制。暴露于exNef的小角质细胞释放Toll样受体诱导的细胞因子和趋化因子如IL-8、IL-6、IL-12、RANTES和IL-17A等因子水平升高。体外实验分别通过跨内皮细胞电阻及右旋糖酐-FITC转运研究测定（表明），含有Nef酰基位点的Nef肽最终穿过血脑屏障降低Nef转染的小胶质细胞释放Nefexosome，并阻止血脑屏障的完整性和通透性的损伤。总的来说，我们发现，exNef从绿色荧光标记的Nef转染的小胶质细胞释放；exNef破坏血脑屏障的完整性和通透性，和血脑屏障密切联系，并诱导小胶质细胞因子/趋化因子的分泌。这些exNef介导的作用明显受Nef肽限制。综上所述，本研究在HIV相关的神经免疫病理机制方面提供了exNef作用的初步证据，以及纳米药为基础的靶向exNef的疗法去治疗HIV相关的神经疾病发病机理的便利性。
Thenegativefactor(Nef)ofhumanimmunodeficiencyvirus(HIV)isanaccessoryproteinthatisthoughttobeintegraltoHIV-associatedimmune-andneuroimmunepathogenesis.Here,weshowthatnef-transfectedmicroglia-releasedNef+exosome(exNef)disruptstheapicalblood-brainbarrier(BBB)andthatonlynef-transfectedmicrogliareleaseNefinexosomes.nef-gfp-transducedneuronsandastrocytesreleaseexosomesbutdidnotreleaseexNefintheextracellularspace.ApicaladministrationofexNefderivedfromnef-transfectedTcellsreducedtransendothelialelectricalresistance(TEER)andincreasedpermeabilityoftheBBB.Microglia-derivedexNefappliedtoeithertheapical/basalBBBsignificantlyreducedexpressionofthetightjunctionprotein,ZO-1,suggestingamechanismofexNef-mediatedneuropathogenesis.MicrogliaexposedtoexNefreleaseelevatedlevelsofToll-likereceptor-inducedcytokinesandchemokinesIL-12,IL-8,IL-6,RANTES,andIL-17A.MagneticnanoparticledeliveryofNefpeptidescontainingtheNefmyrisolationsiteacrossaninvitroBBBultimatelyreducednef-transfectedmicrogliareleaseofNefexosomesandpreventedthelossofBBBintegrityandpermeabilityasmeasuredbyTEERanddextran-FITCtransportstudies,respectively.Overall,weshowthatexNefisreleasedfromnef-gfp-transfectedmicroglia;exNefdisruptsintegrityandpermeability,andtightjunctionsoftheBBB,andinducesmicroglialcytokine/chemokinesecretion.TheseexNef-mediatedeffectsweresignificantlyrestrictedbyNefpeptides.Takentogether,thisstudyprovidespreliminaryevidenceoftheroleofexNefinHIVneuroimmunepathogenesisandthefeasibilityofananomedicine-basedtherapeuticstargetingexNeftotreatHIV-associatedneuropathogenesis.
Nef(negativefactor,Nef)，负调控因子，是人免疫缺陷病毒辅助蛋白之一,相对分子质量小,柔韧性大,核心结构呈球形。
绿色萤光蛋白(greenfluorescentprotein)，简称GFP。
参考文献：Raymond,A.D.,P.Diaz,etal.().Microglia-derivedHIVNef+exosomeimpairmentoftheblood-brainbarrieristreatablebynanomedicine-baseddeliveryofNefpeptides.JNeurovirol22(2):-.
赞赏
人赞赏
鍜ㄨ鐧界櫆椋庝笓瀹?寰椾簡鐧界櫆椋庤鎬庝箞娌荤枟