CN114767844A - 一种水痘-带状疱疹病毒疫苗及其应用 - Google Patents
一种水痘-带状疱疹病毒疫苗及其应用 Download PDFInfo
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Abstract
本发明提供了一种水痘‑带状疱疹病毒疫苗及其应用,属于疫苗技术领域;所述水痘‑带状疱疹病毒疫苗包括脂质体纳米颗粒和包载于所述脂质体纳米颗粒中的带状疱疹病毒糖蛋白E和佐剂;所述佐剂包括三萜皂甙。本发明中,带状疱疹病毒糖蛋白E(gE)被脂质体纳米颗粒包裹后,可有效促进抗原提呈细胞吞噬和高效递送抗原,并实现疫苗的缓释持续刺激机体产生针对VZV‑gE特异的细胞免疫应答。本发明使用的三萜皂甙可有效实现抗原带状疱疹病毒糖蛋白E的交叉提呈,诱导抗原特异性细胞免疫应答。并且,脂质纳米颗粒中富含的胆固醇可有效中和三萜皂甙的细胞毒性,确保该疫苗的安全性。
Description
技术领域
本发明属于疫苗技术领域,特别涉及一种水痘-带状疱疹病毒疫苗及其应用。
背景技术
水痘-带状疱疹病毒(Varicella-Zoster Virus,VZV)遍布全球,具有很强的传染性,迄今只发现一种血清型,在自然界VZV仅感染人类。VZV既可引发水痘,也可引发带状疱疹(herpes zoster,HZ),水痘通常见于儿童期,带状疱症则在成年以后才会发病。在原发感染水痘后,病毒可潜伏在宿主的神经节中,随着年龄增长、免疫功能受损或免疫抑制,VZV可以被重新激活并引发带状疱疹。在全球范围内,绝大多数成年人都有发生带状疱疹及其相关并发症的危险。
由日本人高桥理明(Michiaki Takahashi)开发的Oka株减毒活疫苗1995年被FDA批准用于接种儿童和成人预防水痘(接种量1000-10000PFU(空斑形成单位,plaqueforming unit)),之后广泛应用于全世界。后续的研究发现,Oka株和野生型病毒一样会建立潜伏感染,进而同样可能导致带状疱疹的发生。
对既往感染VZV病毒的50岁以上人群一次性皮下加强免疫高剂量减毒活疫苗(接种量约19400PFU)可有效预防带状疱疹,默克公司(Merk)相应产品Zostavax 2005年上市,对50~59,60~69以及70岁以上人群的保护率分别为70%,64%和38%左右。这种保护率随年龄增长的降低主要归因于伴随免疫系统老化出现的细胞免疫应答削弱。
葛兰素史克(GSK)于2017年底上市的带状疱疹基因工程亚单位疫苗Shingrix使用中国仓鼠卵巢细胞(Chinese hamster ovary,CHO)表达的保守性病毒糖蛋白E(gE)作为抗原,使用佐剂AS01B有效增强针对VZV-gE特异的细胞免疫应答,使其在50岁以上健康人群中的保护率高达97.2%(对50~59,60~69和70岁以上人群保护率分别为96.6%,97.3%和91.3)且在包括HIV携带者在内的免疫缺陷人群中表现出了良好的安全性和有效性。AS01B佐剂系统中的三萜多糖QS21和单磷酰脂质(MPL)A基于脂质体载体发挥协同作用,诱导针对gE特异性的CD4阳性T细胞,对疫苗效果发挥关键作用。
将含有CpG基序的寡脱氧核苷酸(CpG ODN)封装到可电离脂质纳米颗粒(LNP)中,可增强抗原特异性体液免疫应答和细胞免疫应答(PMID:33805880)。
由于QS21存在强细胞毒性使许多研究未得到临床应用。因此如何在保证疫苗组分安全性的前提下,使用合适佐剂成分增强VZV-gE特异性细胞免疫应答,获得类似Shingrix带状疱疹疫苗的免疫效果,是疫苗开发亟需解决的问题。
发明内容
有鉴于此,本发明的目的在于提供一种水痘-带状疱疹病毒疫苗及其应用,本发明的水痘-带状疱疹病毒疫苗可有效增强针对VZV-gE特异的细胞免疫应答,用作带状疱疹疫苗。并且本发明的水痘-带状疱疹病毒疫苗安全性高。
本发明提供了一种水痘-带状疱疹病毒疫苗,包括脂质体纳米颗粒和包载于所述脂质体纳米颗粒中的带状疱疹病毒糖蛋白E和佐剂;所述佐剂包括三萜皂甙。
优选的,所述水痘-带状疱疹病毒疫苗中带状疱疹病毒糖蛋白E的含量为5~100μg/剂。
优选的,所述水痘-带状疱疹病毒疫苗中三萜皂甙的含量为1~100μg/剂。
优选的,所述三萜皂苷包括QS21。
优选的,所述佐剂还包括富含GC的单链寡聚脱氧核苷酸片段。
优选的,所述水痘-带状疱疹病毒疫苗中含GC的单链寡聚脱氧核苷酸片段的含量为5μg~2mg/剂。
优选的,所述脂质体纳米颗粒包含阳离子脂质体和聚乙二醇衍生物;所述阳离子脂质体和聚乙二醇衍生物的摩尔比为(46~50):(1.5~1.6)。
优选的,所述水痘-带状疱疹病毒疫苗的粒径为20~400nm。
优选的,所述水痘-带状疱疹病毒疫苗的剂型包括注射剂。
本发明还提供了上述方案所述的水痘-带状疱疹病毒疫苗在制备预防或改善带状疱疹和/或带状疱疹后遗症的药物中的应用。
本发明提供了一种水痘-带状疱疹病毒疫苗,包括脂质体纳米颗粒和包载于所述脂质体纳米颗粒中的带状疱疹病毒糖蛋白E和佐剂;所述佐剂包括三萜皂甙。本发明中,带状疱疹病毒糖蛋白E(gE)被脂质体纳米颗粒包裹后,可有效促进抗原提呈细胞吞噬和高效递送抗原,并实现疫苗的缓释持续刺激机体产生针对VZV-gE特异的细胞免疫应答。本发明使用的三萜皂甙可有效实现抗原带状疱疹病毒糖蛋白E的交叉提呈,诱导抗原特异性细胞免疫应答。并且,脂质纳米颗粒中富含的胆固醇可有效中和三萜皂甙的细胞毒性,确保该疫苗的安全性。本发明所述的水痘-带状疱疹病毒疫苗经动物实验证实能特异性增强针对带状疱疹病毒糖蛋白E的细胞免疫应答,可用作带状疱疹疫苗。
附图说明
图1-显示经实施例制备的水痘-带状疱疹病毒疫苗,经实验例1检测所得抗原包封率(图1中的A),经实验例2检测所得核酸包封率(图1中的B),经实验例3检测所得QS21包封率(图1中的C),经实验例4检测所得粒径(图1中的D),经实验例4所得多分散性指数(图1中的E);
图2-显示经实施例制备的水痘-带状疱疹病毒疫苗,经实验例5检测所得细胞毒性;
图3-显示经实施例制备的水痘-带状疱疹病毒疫苗,经实验例6、实验例7、实验例12检测所得gE特异性IgG抗体滴度;
图4-显示经实施例制备的水痘-带状疱疹病毒疫苗,经实验例6、实验例8、实验例9、实验例12检测所得IL-2浓度;
图5-显示经实施例制备的水痘-带状疱疹病毒疫苗,经实验例6、实验例8、实验例9、实验例12检测所得IFN-γ浓度;
图6-显示经实施例制备的水痘-带状疱疹病毒疫苗,经实验例6、实验例8、实验例10、实验例12检测所得每2×105个脾细胞分泌IL-2形成的斑点数;
图7-显示经实施例制备的水痘-带状疱疹病毒疫苗,经实验例6、实验例8、实验例10、实验例12检测所得每2×105个脾细胞分泌IFN-γ形成的斑点数;
图8-显示经实施例制备的水痘-带状疱疹病毒疫苗,经实验例6、实验例8、实验例11、实验例12检测所得分泌IL-2的CD4+T细胞比例;
图9-显示经实施例制备的水痘-带状疱疹病毒疫苗,经实验例6、实验例8、实验例11、实验例12检测所得分泌IFN-γ的CD4+T细胞比例。
具体实施方式
本发明提供了一种水痘-带状疱疹病毒疫苗,包括脂质体纳米颗粒和包载于所述脂质体纳米颗粒中的带状疱疹病毒糖蛋白E和佐剂;所述佐剂包括三萜皂甙。
在本发明中,所述水痘-带状疱疹病毒疫苗中各组分通过物理电吸附或物理包裹结合。
在本发明中,所述水痘-带状疱疹病毒疫苗中带状疱疹病毒糖蛋白E的含量优选为5~100μg/剂。本发明中,带状疱疹病毒糖蛋白E(gE)被脂质体纳米颗粒包裹后,可有效促进抗原提呈细胞吞噬和高效递送抗原,并实现疫苗的缓释持续刺激机体产生针对VZV-gE特异的细胞免疫应答。
在本发明中,所述水痘-带状疱疹病毒疫苗中三萜皂甙的含量优选为1~100μg/剂。在本发明中,所述三萜皂苷优选的包括QS21,提取自南美皂角树Quillaja saponaria树皮。本发明使用的三萜皂甙可有效实现抗原带状疱疹病毒糖蛋白E的交叉提呈,诱导抗原特异性细胞免疫应答。并且,脂质纳米颗粒中富含的胆固醇可有效中和三萜皂甙的细胞毒性,确保该疫苗的安全性。
在本发明中,所述佐剂优选的还包括富含GC的单链寡聚脱氧核苷酸片段(CpGODN),更优选为CpG ODN 1018。在本发明中,所述水痘-带状疱疹病毒疫苗中含GC的单链寡聚脱氧核苷酸片段的含量优选为5μg~2mg/剂。在本发明中,CpG ODN由脂质纳米颗粒包裹,一方面有效避免核酸酶的降解;另一方面,在被提呈细胞吞噬前逃逸出的CpG ODN可迅速被体内的核酸酶降解,从而有效避免了CpG ODN从疫苗注射部位非特异性扩散可能引起的系统性炎症副作用,使佐剂呈现“局域性”和“一过性”的特点,符合安全性要求。此外,本发明所述的水痘-带状疱疹病毒疫苗,使用的CpG ODN可被分布于内吞体内的TLR9摄取,诱导干扰素分泌,并通过促进抗原的交叉提呈有效活化抗原特异性T细胞。其中CpG ODN的A类可刺激树突状细胞产生I型干扰素、活化自然杀伤细胞,B类可快速从早期内体转移至晚期内体、刺激B细胞增殖、刺激浆细胞样树突状细胞成熟和TNF-α、IL-6及IL-12的产生,C类兼具A类和B类的作用特点、平衡促进体液免疫和细胞免疫应答。逃逸至细胞质中的可形成局部茎环结构的C型CpG ODN则可能通过激活cyclic GMP-AMP synthase(cGAS)继而通过stimulatorof IFN genes(STING)天然免疫通路,诱导相关获得性免疫应答。本发明同时使用三萜皂甙与CpG ODN,两者在诱导抗原特异的细胞免疫应答方面具有很好的协同作用。
在本发明中,所述脂质体纳米颗粒包含阳离子脂质体和聚乙二醇衍生物;所述阳离子脂质体和聚乙二醇衍生物的摩尔比为(46~50):(1.5~1.6)。在本发明中,所述阳离子脂质体优选的包括((4-羟基丁基)氮杂二烷基)双(己烷-6,1-二基)双(2-己基癸酸酯)(ALC-0315)和/或十七烷-9-基-8-((2-羟乙基)(6-氧代-6-((十一烷氧基)己基)氨基)辛酸酯)(SM-102)。在本发明中,所述聚乙二醇衍生物优选的包括甲氧基聚乙二醇双十四烷基乙酰胺(ALC-0159)和1,2-二肉豆蔻酰-rac-甘油-3-甲氧基聚乙二醇(DMG-PEG2000)。
在本发明中,所述水痘-带状疱疹病毒疫苗的粒径优选为20~400nm。
在本发明中,所述水痘-带状疱疹病毒疫苗的剂型优选的包括注射剂。
本发明对所述水痘-带状疱疹病毒疫苗的制备方法没有特殊限制,采用本领域常规的质体纳米颗粒包覆方法即可,本发明具体实施过程中,优选的采用微流控技术设备制备水痘-带状疱疹病毒疫苗。
在本发明中,所述水痘-带状疱疹病毒疫苗的给药方式优选为注射给药;所述注射优选的包括皮下注射或肌肉注射。
本发明还提供了上述方案所述的水痘-带状疱疹病毒疫苗在制备预防或改善带状疱疹和/或带状疱疹后遗症的药物中的应用。在本发明中,所述带状疱疹后遗症优选的包括带状疱疹后神经痛。
下面将结合本发明中的实施例,对本发明中的技术方案进行清楚、完整地描述。
按照下表1中单针剂疫苗组成,计算20针剂各疫苗组所需组分投料。
表1.单针剂疫苗投料
-未加入;√已加入.
对比例1—称取CHO表达的gE胞外区糖蛋白(购自昆明滇工科技有限公司)0.2mg,0.2mg硫代氧化CpG 1018(购自InvivoGen公司),0.1mgQS21(购自Alpha Diagnostic公司),溶解于1mLPBS,即得对比例1疫苗20针剂。
对比例2—称取CHO表达的gE胞外区糖蛋白0.2mg,0.2mg硫代氧化CpG 1018,0.1mgQS21,溶解于0.5mLPBS,与等体积铝佐剂(购自Thermo Fisher公司)混匀,即得对比例2疫苗20针剂。
实施例—按照ALC-0315(购自厦门赛诺邦格生物科技股份有限公司):DSPC(购自上海艾韦拓医药科技有限公司):cholesterol(购自上海艾韦拓医药科技有限公司):ALC-0159(购自厦门赛诺邦格生物科技股份有限公司)摩尔比为46.3:9.4:42.7:1.6称取脂质后溶解于无水乙醇,使用微流体纳米药物制造系统(Precision Nanosystems公司)按照1:3比例混合溶解于含0.24mg gE、0.3mg CpG 1018和0.12mg QS21的100mM、pH 4.0的柠檬酸缓冲液,即得实施例疫苗20针剂。
针对以上实施例及对比例1~2制备得到的疫苗,进行如下实验测定:
实验例1、gE浓度
实施例疫苗于0.1M氢氧化钠和0.1%(w/v)的十二烷基硫酸钠缓冲液室温裂解过夜。使用BCA比色法蛋白检测试剂盒(上海碧云天生物技术有限公司)检测gE浓度并计算蛋白荷载效率。
实验例2、核酸浓度
实施例疫苗于0.1M氢氧化钠和0.1%(w/v)的十二烷基硫酸钠缓冲液室温裂解过夜。使用核酸检测试剂盒Quant-iT OliGreen ssDNA Regent Kit(购自Thermo Fisher公司)检测核酸浓度并计算核酸荷载效率。
实验例3、QS21浓度
实施例疫苗于0.1M氢氧化钠和0.1%(w/v)的十二烷基硫酸钠缓冲液室温裂解过夜。以游离QS21为标准品,使用4.6×250mm C18柱(购自Waters公司)的高效液相色谱(HPLC,购自Waters公司)对实施例中所包裹的QS21含量进行测定并计算QS21荷载效率。
实验例4、粒径和多分散性指数
实施例疫苗使用纳米粒径检测仪(马尔文)检测LNP的粒径和多分散性指数。
实施例和实验例1~4结果如图1所示。实施例使用微流体纳米药物制造系统制备的LNP脂质纳米疫苗gE的包封率为49.57%(图1中的A),约得5.95μg/针剂;CpG ODN核酸的包封率为41.85%(图1中的B),约得6.28μg/针剂;QS21的荷载效率为57.15%(图1中的C),约得3.43μg/针剂;纳米颗粒粒径在190.3~194.7nm之间(图1中的D),多分散性指数在0.269~0.322之间(图1中的E)。
实验例5、细胞毒性
取无特定病原体C57BL/6J小鼠(雌性,6~8周龄,16~18g,购自成都达硕实验动物有限公司)的股骨和胫骨,ACK红细胞裂解液裂解红细胞获取骨髓细胞。使用含有20ng/mLGM-CSF(购自苏州派普泰克生物科技有限公司)的1640完全培养基(购自Thermo Fisher公司)诱导未成熟的骨髓来源树突状细胞(BMDCs)。2×105个细胞每孔接种96孔板后,加入样品继续培养24h。使用CCK-8试剂盒(购自MedChemExpress)检测细胞活力。
实施例和实验例5结果如图2所示。10μg/mL对比例游离QS21存在时BMDC细胞活性仅为约18%。当用LNP包裹后,实施例中相同浓度的QS21并没有显示出明显的细胞毒性。
实验例6、动物免疫
以PBS为空白对照,间隔4周,肌肉注射实施例、对比例1、对比例2制备的疫苗50μl免疫C57BL/6小鼠2次(6只/组,雌性,初免年龄6~8周,体重16~18g),终免2周后摘取脾脏,心脏取血4℃放置过夜后3500转/min离心30min取血清,准备进行后续免疫学分析。
实验例7、抗体滴度检测
溶解于PBS的捕获抗原gE胞外区糖蛋白2μg/mL每孔100μL加入96孔酶标板(购自康宁),4℃过夜包被后PBST(0.05%(v/v)Tween20(Sigma)in PBS)洗板1次,每孔200μl加入溶于PBS的5%(w/v)脱脂奶粉封闭液37℃封闭1h,弃掉封闭液后PBST洗4次,每孔100μl加入1%封闭液梯度稀释的抗血清37℃孵育1h,PBST洗5次后加入1%封闭液稀释的二抗(1:10000,Goat anti-mouse IgG:HRP购自BioRad)37℃孵育1h,PBST洗5次后每孔加入100μl按照1:1比例配好的显色液(购自BD),室温下避光放置5min后,加入100μl 1M硫酸终止反应,并于450nm处检测光吸收值。取OD450>0.15临界的血清稀释浓度作为抗体滴度,在1:2000的稀释度下OD450小于0.15的滴度定义为100用于计算。
对比例1~2、实施例、实验例6、7结果如图3所示。实施例免疫小鼠血清中的gE特异性IgG滴度为170667,与对比例2相当,是对比例1的1.3倍(IgG滴度为128000)。
实验例8、脾脏淋巴细胞分离
脾脏置细胞过滤网(购自无锡耐思生命科技股份有限公司),加入ACK红细胞裂解液室温放置5min,1800转/min离心后细胞计数,并使用含有10%胎牛血清(购自ThermoFisher)和双抗的1640培养基(购自Thermo Fisher)重悬为1×107细胞/mL。
实验例9、细胞因子分析
将100μl 1×107细胞/mL的脾细胞添加到96孔板(购自康宁)的每个孔中。每孔加入终浓度为10μg/mL的gE,并使用10μLPMA+离子霉素(原液浓度:500ng/mL+10μg/mL;购自达科为)作为阳性对照。37℃、5%CO2环境下孵育24h后,收集细胞上清液,ELISA法检测IL-2和IFN-γ的含量。将溶解在PBS中的IL-2(3μg/mL)和IFN-γ(4μg/mL)捕获抗体(购自ThermoFisher)于4℃下包被96孔板16h。用5%脱脂牛奶封闭液37℃封闭1h后,每孔加入50μl细胞上清液,室温孵3h。PBS溶解的小鼠IL-2和IFN-γ蛋白标准品(购自苏州派普泰克生物科技有限公司)用于生成标准曲线。随后添加对IL-2或IFN-γ特异的生物素偶联抗体(2μg/mL,购自Thermo Fisher)和HRP偶联链霉抗生物素蛋白(1μg/mL,购自BioLegend)并孵育1.5h。如抗体滴度检测中所述终止并检测反应结果。
对比例1~2、实施例、实验例6、8、9、12结果如图4~5所示。通过ELISA分析,实施例上清液中的IL-2水平为2509pg/mL(图4)。该水平是对比例1的1.25倍(2011pg/mL,p=0.72),是对比例2的2.69倍(934.3pg/mL,p=0.02)。实施例上清液中的IFN-γ水平为6722pg/mL(图5)。该水平是对比例1的1.21倍(5572pg/mL,p=0.42),是对比例2的1.6倍(4207pg/mL,p=0.02)。
实验例10、酶联免疫斑点实验(enzyme linked immunospot assay,ELISPOT)
IL-2及IFN-γ检测试剂盒均购自BD,并按照说明书进行操作,具体步骤如下:包被液稀释捕获抗体后100μL/孔加入ELISPOT平板,4℃包被过夜后弃包被液,200μL/孔封闭液洗板1次,每孔200μL加入封闭液室温封闭2h,弃封闭液后加入含有20μg/mL终浓度gE的1640完全培养基100μL并加入上述脾脏淋巴细胞分离中得到的脾脏细胞使终浓度为2×105个细胞/孔,37℃细胞培养箱过夜。800g离心5min后弃上清,200μL/孔去离子水清洗2次(每次浸泡5min),200μL/孔清洗液1清洗3次,100μL/孔加入经稀释液稀释的检测抗体室温孵育2h后,200μL/孔清洗液1清洗3次(每次浸泡2min),100μL/孔加入经稀释液稀释的酶偶合物Streptavidin-HRP室温孵育1h,200μL/孔清洗液1清洗4次(每次浸泡2min),200μL/孔清洗液2清洗2次后加入100μL底物溶液反应至合适时间,去离子水清洗终止反应。晾干后使用ELISPOT读板仪器(AID Diagnostika GmbH)斑点计数。
对比例1~2、实施例、实验例6、8、10、12结果如图6~7所示。通过ELISPOT分析,实施例中gE刺激后分泌IL-2的细胞数量为224.3/2×105个脾细胞(图6)。该数字是对比例1的2.1倍(每2×105个脾细胞106.8个,p<0.001),是对比例2的1.87倍(每2×105个脾细胞119.8个,p=0.002)。在实施例中gE刺激后IFN-γ分泌细胞的数量为每2×105个脾细胞293.7个(图7)。该数字是对比例1的1.67倍(每2×105个脾细胞175.5个,p=0.008),是对比例2的1.87倍(每2×105个脾细胞157个,p=0.002)。
实验例11、流式分析
所有流式分析试剂均购自BioLegend。将总共2×106个脾细胞与10μg/mL蛋白gE在37℃、5%CO2中孵育2h,然后加入5μg/mL布雷菲德菌素A。脾细胞在相同条件下孵育过夜以阻断细胞因子释放。用染色缓冲液洗涤后,将100μl Zombie NIRTM添加到每个样品中并孵育30min。然后加入5μg/ml抗CD16/CD32抗体,将脾细胞在4℃孵育10min,以阻断与Fc受体的非特异性结合。随后加入PerCP-Cy5.5-缀合的抗小鼠CD4并在4℃孵育30min。PE缀合的抗小鼠IFN-γ和APC缀合的抗小鼠IL-2抗体用于细胞内染色。染色后,对细胞进行门控(前向和侧向散射,FSC/SSC)并用CytoFLEX流式细胞仪(Beckman)和FlowJo_V10软件分析超过20000个CD4+细胞事件的样品。
对比例1~2、实施例、实验例6、8、11、12结果如图8~9所示。根据流式细胞仪分析,在实施例中gE刺激后表达IL-2的CD4+T细胞的比例为0.6633%(图8)。该水平是对比例1的2.54倍(0.2612%,p=0.008),是对比例2的3.04倍(0.2183%,p=0.004)。实施例中gE刺激后表IFN-γ的CD4+T细胞比例为0.7598%(图9)。该水平是对比例1的2.11倍(0.3598%,p=0.04),是对比例2的3.53倍(0.2152%,p=0.004)。
实验例12、统计分析
使用GraphPad Prism 9.2软件分析数据,并表示为平均值±SD。通过普通单向方差分析(ANOVA)和Dunnett′s多重比较检验以实施例为基准分析实验组之间的显着差异。星号代表p值分类:*p≤0.05、**p≤0.01和***p≤0.001。
实验例12结果如图4~9所示。
尽管上述实施例对本发明做出了详尽的描述,但它仅仅是本发明一部分实施例而不是全部实施例,人们还可以根据本实施例在不经创造性前提下获得其他实施例,这些实施例都属于本发明保护范围。
Claims (10)
1.一种水痘-带状疱疹病毒疫苗,包括脂质体纳米颗粒和包载于所述脂质体纳米颗粒中的带状疱疹病毒糖蛋白E和佐剂;所述佐剂包括三萜皂甙。
2.根据权利要求1所述的水痘-带状疱疹病毒疫苗,其特征在于,所述水痘-带状疱疹病毒疫苗中带状疱疹病毒糖蛋白E的含量为5~100μg/剂。
3.根据权利要求1或2所述的水痘-带状疱疹病毒疫苗,其特征在于,所述水痘-带状疱疹病毒疫苗中三萜皂甙的含量为1~100μg/剂。
4.根据权利要求1所述的水痘-带状疱疹病毒疫苗,其特征在于,所述三萜皂苷包括QS21。
5.根据权利要求1或4所述的水痘-带状疱疹病毒疫苗,其特征在于,所述佐剂还包括富含GC的单链寡聚脱氧核苷酸片段。
6.根据权利要求5所述的水痘-带状疱疹病毒疫苗,其特征在于,所述水痘-带状疱疹病毒疫苗中含GC的单链寡聚脱氧核苷酸片段的含量为5μg~2mg/剂。
7.根据权利要求1所述的水痘-带状疱疹病毒疫苗,其特征在于,所述脂质体纳米颗粒包含阳离子脂质体和聚乙二醇衍生物;所述阳离子脂质体和聚乙二醇衍生物的摩尔比为(46~50):(1.5~1.6)。
8.根据权利要求1所述的水痘-带状疱疹病毒疫苗,其特征在于,所述水痘-带状疱疹病毒疫苗的粒径为20~400nm。
9.根据权利要求1所述的水痘-带状疱疹病毒疫苗,其特征在于,所述水痘-带状疱疹病毒疫苗的剂型包括注射剂。
10.权利要求1~9任意一项所述的水痘-带状疱疹病毒疫苗在制备预防或改善带状疱疹和/或带状疱疹后遗症的药物中的应用。
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