CN113855796B - 卡介苗作为呼吸道合胞病毒灭活疫苗佐剂的应用 - Google Patents
卡介苗作为呼吸道合胞病毒灭活疫苗佐剂的应用 Download PDFInfo
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Abstract
本发明公开了一种卡介苗作为呼吸道合胞病毒灭活疫苗佐剂的应用，BCG作为RSV灭活疫苗的佐剂，BCG诱导的Th1型细胞免疫应答可以平衡RSV灭活疫苗所诱导的过强的Th2和Th17型细胞免疫应答，使疫苗接种者体内产生平衡的免疫记忆，当RSV感染时，在提供保护的同时防止疫苗增强性炎症疾病的发生，即既安全又有效。
Description
技术领域
本发明涉及疫苗及佐剂技术领域，具体的涉及卡介苗作为呼吸道合胞病毒灭活疫苗佐剂的应用。
背景技术
呼吸道合胞病毒(Respiratory syncytial virus,RSV)是婴幼儿下呼吸道感染的最重要病原，几乎所有的儿童在两岁之前都被感染过，在5岁以下儿童中，RSV感染需要急诊和住院治疗的病例数是流感病毒感染的十多倍，是婴幼儿住院和死亡的最重要原因；另外，老年人也是RSV的易感人群，每年的感染率与流感相似，而死亡率几乎是流感的2倍。全球每年因RSV产生的直接经济负担超过800亿美元，而至今仍无安全有效的疫苗上市，因此，WHO将RSV疫苗列为优先发展的疫苗。
自1956年发现RSV后不久，疫苗的研发工作就迅速开展起来了，不幸的是1960年代辉瑞公司研制的***灭活RSV疫苗(FI-RSV)在临床试验中出现了重大挫折：FI-RSV被批准在婴幼儿中进行临床试验，接种对象是两个月-七岁的幼儿，临床试验进行9个月后进入RSV流行季节，接种FI-RSV疫苗的婴幼儿不仅未被保护，反而在RSV感染后发生了比安慰剂组更严重的肺部炎症疾病，肺部出现了以大量中性粒细胞和嗜酸性粒细胞浸润及Th2型免疫应答为主的过敏样炎症，即疫苗增强性疾病(vaccine-enhanced disease，VED)，导致疫苗接种者中80％的患儿需要住院治疗，其中两例死亡，而安慰剂组仅5％需要住院治疗[1]。该疫苗的失败严重打击了RSV疫苗的研发热情，增加了RSV疫苗研发的商业风险，“安全性”成为RSV疫苗研究者首要考虑的问题。
动物实验的研究显示：FI-RSV疫苗增强性疾病的发病机制主要与疫苗诱导的不平衡免疫应答有关[2]，特别是Th2和Th17型细胞免疫优势应答，而缺乏Th1型细胞免疫应答[3]。
佐剂是指预先或与抗原同时注入体内，可增强机体对抗原的免疫应答或改变免疫应答类型的非特异性免疫增强性物质。铝盐佐剂(如Al(OH)3和磷酸铝等)是已被使用了近百年的人用疫苗佐剂，绝大多数疫苗使用的佐剂为此种类型。
卡介苗(BCG)是由减毒牛型结核杆菌悬浮液制成的活菌苗，可预防结核和麻风病等分枝杆菌感染，现已使用了近一个世纪。另外，BCG具有增强巨噬细胞活性，诱导Th1型免疫应答的特征，是一类促进Th1型免疫应答的良好佐剂。
另有证据表明BCG可以保护婴儿免受除结核分枝杆菌以外的病原体的感染，从而提供异源或非特异***叉保护[4]。有文献表明，卡介苗可减少其他呼吸道感染、腹泻或疟疾造成的死亡；来自几内亚比绍和巴西的小型研究显示，接种过BCG疫苗的儿童患肺炎的比率较低；同样，最近发表的一项基于国际人口和健康调查的研究表明，接种卡介苗可使疑似急性下呼吸道感染的风险降低17％至37％，该调查包括来自37个国家的15万名儿童；卡介苗不仅可以保护儿童免受呼吸道感染，还可以降低其严重程度和相关死亡率。
参考文献：
1.Kapikian AZ,Mitchell RH,Chanock RM,Shvedoff RA,Stewart CE.Anepidemiologic study of altered clinical reactivity to respiratory syncytial(RS)virus infection in children previously vaccinated with an inactivated RSvirus vaccine.Am J Epidemiol 1969；89(4):405-421.
2.Ruckwardt TJ,Morabito KM,Graham BS.Immunological Lessons fromRespiratory Syncytial Virus Vaccine Development.Immunity 2019；51(3):429-442.
3.Zhang L,Li H,Hai Y,Yin W,Li W,Zheng B,Ruihong Zeng.CpG inCombination with an Inhibitor of Notch Signaling Suppresses Formalin-Inactivated Respiratory Syncytial Virus-Enhanced Airway Hyperresponsivenessand Inflammation by Inhibiting Th17 Memory Responses and Promoting Tissue-Resident Memory Cells in Lungs.JVirol 2017；91(10).
4.Arts RJW,Moorlag S,Novakovic B,Li Y,Wang SY,Oosting M,et al.BCGVaccination Protects against Experimental Viral Infection in Humans throughthe Induction of Cytokines Associated with Trained Immunity.Cell Host Microbe2018；23(1):89-100e105
现有技术中，还没有以卡介苗作为呼吸道合胞病毒灭活疫苗佐剂的相关报道和研究。
发明内容
针对现有技术存在的上述问题，本发明提供了卡介苗作为呼吸道合胞病毒灭活疫苗佐剂的应用，BCG作为RSV灭活疫苗的佐剂，BCG诱导的Th1型细胞免疫应答可以平衡RSV灭活疫苗所诱导的过强的Th2和Th17型细胞免疫应答，使疫苗接种者体内产生平衡的免疫记忆，当RSV感染时，在提供保护的同时防止疫苗增强性炎症疾病的发生，即实现既安全又有效。
为实现上述技术目的，达到上述技术效果，本发明是通过以下技术方案实现：
卡介苗(BCG)作为制备治疗或预防呼吸道合胞病毒(Respiratory syncytialvirus，RSV)灭活疫苗佐剂的的用途，灭活疫苗为***灭活RSV疫苗FI-RSV。
进一步的，所述灭活疫苗含或不含铝盐佐剂。
进一步的，所述用途为将BCG提前或同时与RSV灭活疫苗注射体内，诱导平衡的免疫记忆。
进一步的，所述用途为佐剂BCG仅注射一次或与灭活RSV疫苗注射相同次数。
本发明的有益效果：
本发明提供了卡介苗作为呼吸道合胞病毒灭活疫苗佐剂的应用，BCG作为RSV灭活疫苗的佐剂，BCG诱导的Th1型细胞免疫应答可以平衡RSV灭活疫苗所诱导的过强的Th2和Th17型细胞免疫应答，使疫苗接种者体内产生平衡的免疫记忆，当RSV感染时，在提供保护的同时防止疫苗增强性炎症疾病的发生，即实现既安全又有效。
当然，实施本发明的任一产品并不一定需要同时达到以上所述的所有优点。
附图说明
为了更清楚地说明本发明实施例的技术方案，下面将对实施例描述所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1为本发明实施例所述RSV灭活疫苗+佐剂免疫乳鼠并经RSV攻击后各组肺组织病理变化；A、PBS；B、PBS/RSV；C、FI-RSV+Al(OH)3；D、FI-RSV+BCG(two)；E、FI-RSV+BCG(one)；F、FI-RSV+Al(OH)3+BCG(one)；
图2为本发明实施例所述RSV灭活疫苗+佐剂免疫乳鼠并经RSV攻击后用实时定量PCR检测各组肺组织中RSV-N蛋白及细胞因子mRNA的表达(*:p<0.05,**:p<0.01，***：p<0.001)；
图3为本发明实施例所述RSV灭活疫苗+佐剂免疫成年小鼠并经RSV攻击后各组肺组织病理变化(H&E染色100×)；A、PBS；B、PBS/RSV；C、FI-RSV+Al(OH)3；D、FI-RSV+BCG；E、FI-RSV+Al(OH)3+BCG；
图4为本发明实施例所述RSV灭活疫苗+佐剂免疫成年小鼠并经RSV攻击后用实时定量PCR检测各组肺组织中RSV-N蛋白及细胞因子mRNA的表达(*:p<0.05,**:p<0.01，***：p<0.001)；
具体实施方式
下面将结合本发明实施例中的附图，对本发明实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其它实施例，都属于本发明保护的范围。
实施例1
用RSV灭活疫苗+佐剂免疫乳鼠的实验研究
1.1乳鼠分组：
①PBS组；
②PBS/RSV组；
③FI-RSV+Al(OH)3组；
④FI-RSV+BCG(two)组；
⑤FI-RSV+BCG(one)；
⑥FI-RSV+Al(OH)3+BCG(one)。6-10只/组
1.2乳鼠的疫苗接种及RSV攻击方案：
注：i.m.肌内注射，s.c.皮下注射。
领取BALB/c孕鼠，孕鼠生产后，乳鼠出生当天记为0天，皮下接种BCG(④-⑥组)，出生1天肌肉注射PBS(①和②组)、FI-RSV(④和⑤组)或FI-RSV+Al(OH)3(③和⑥组)，第7天④组再次皮下接种BCG，第8天再次肌肉注射PBS、FI-RSV或FI-RSV+Al(OH)3，第21天用30微升PBS(①组)或RSV(②-⑥组)滴鼻攻击，5天后处死，取肺组织，石蜡包埋，切片，H&E染色；取部分肺组织，提取RNA，进行实时定量PCR检测。
实施例2
用RSV灭活疫苗+佐剂免疫成年小鼠的实验研究
2.1成年小鼠的分组
①PBS组、②PBS/RSV组、③FI-RSV+Al(OH)3组、④FI-RSV+BCG组、⑤FI-RSV+Al(OH)3+BCG组。6只/组
2.2成年小鼠疫苗接种及RSV攻击方案：
注：i.m.是肌内注射，s.c.是皮下注射。
领取6周雌性BALB/c小鼠，皮下接种BCG(④和⑤组)，1天后肌肉注射PBS(①和②组)、FI-RSV(④组)或FI-RSV+Al(OH)3(③和⑤组)，第11天和21天再次肌肉注射PBS、FI-RSV或FI-RSV+Al(OH)3，第35天用50微升PBS(①组)或RSV(②-⑤组)滴鼻攻击，5天后处死，取肺组织，石蜡包埋，切片，H&E染色；取部分肺组织，提取RNA，进行实时定量PCR检测。
统计结果分析
1.RSV攻击后各组乳鼠肺组织病理、RSV-N蛋白及细胞因子表达
注：PBS组：既不接种疫苗，也不用RSV攻击感染；PBS/RSV组：不接种疫苗，但用RSV攻击感染；FI-RSV+BCG(two)组：BCG注射两次；FI-RSV+BCG(one)和FI-RSV+Al(OH)3+BCG(one)组：BCG仅注射一次。
肺组织病理切片，H&E染色结果(如图1)：A、PBS组：正常组可见肺泡结构完整，肺泡间隔正常，支气管柱状上皮细胞平滑。B、PBS/RSV组：局部有炎性细胞浸润，部分肺泡发生融合，肺泡变大，支气管周围发生了炎细胞的浸润。C、FI-RSV+Al(OH)3组：支气管周围有明显的炎症细胞浸润，肺泡间隔增厚，肺泡融合，支气管柱状细胞向内突出，表明出现了明显的疫苗增强性肺部炎症。D、FI-RSV+BCG(two)：炎症细胞浸润有所减轻，肺泡间隔趋于正常，支气管周围存在炎症细胞浸润，有明显的充血水肿，有部分肺泡存在融合，肺泡变大，炎症较FI-RSV+Al(OH)3组明显减轻，但与FI-RSV+BCG(one)、FI-RSV+Al(OH)3+BCG(one)相比，炎症症状较重。E、FI-RSV+BCG(one)：炎症细胞浸润明显减轻，但依旧存在明显的水肿，肺泡间隔趋于正常，有部分肺泡存在融合，肺泡变大，但炎症较FI-RSV+Al(OH)3组明显减轻。F、FI-RSV+Al(OH)3+BCG(one)：没有明显的炎症细胞浸润，没有明显的充血水肿，肺泡结构完整，肺泡间隔相对正常，支气管柱状上皮形态相对正常。
病理切片结果提示：FI-RSV+Al(OH)3组显示了明显的疫苗增强性炎症，而FI-RSV+Al(OH)3+BCG(one)组的炎症明显受到抑制，为最佳免疫接种方案。
如图2所示
与FI-RSV+Al(OH)3组相比FI-RSV+Al(OH)3+BCG(one)组的肺组织中RSV-N(RSV病毒的看家基因-N基因)的表达水平显著降低，表明FI-RSV+Al(OH)3+BCG(one)组的免疫接种方案能够提供很好的保护性；
另外，FI-RSV+Al(OH)3+BCG(one)组的肺组织中eotaxin(嗜酸性粒细胞趋化因子)、Gro-α(中性粒细胞趋化因子)、MCP-1(单核细胞趋化因子)、TNF-α(促炎症细胞因子)、IL-17(Th17型炎症细胞因子)、IL-6和IL-10(Th2型细胞因子)的表达水平明显低于FI-RSV+Al(OH)3组，而FI-RSV+Al(OH)3+BCG(one)组的肺组织中IFN-γ(Th1型细胞因子，可活化巨噬细胞和淋巴细胞，增强其杀伤病原体的能力)的表达水平比PBS及PBS/RSV组均明显增高，与FI-RSV+Al(OH)3组相比无统计学差异。这些结果表明：FI-RSV+Al(OH)3+BCG(one)免疫接种乳鼠后提供了明显的保护，而且肺部的Th2和Th17细胞因子和促炎症因子的水平明显降低。
此部分实验结果提示：对于新生儿，FI-RSV+Al(OH)3+BCG(one)为最佳免疫接种方案，既安全又有效。
2.RSV攻击后各组成年小鼠肺组织病理、RSV-N蛋白及细胞因子表达
肺组织病理切片，H&E染色结果(如图3)，PBS组：正常组可见肺泡结构完整，肺泡间隔正常，支气管柱状上皮细胞平滑。B、PBS/RSV组：局部有炎性细胞浸润，部分肺泡发生融合，肺泡变大，支气管周围发生了炎细胞的浸润。C、FI-RSV+Al(OH)3组：支气管周围有明显的炎症细胞浸润，肺泡间隔增厚，肺泡融合，支气管柱状细胞向内突出，显示明显的疫苗增强性肺部炎症。D、FI-RSV+BCG组：未见明显的炎症细胞浸润，肺泡间隔正常，偶有肺泡存在融合。E、FI-RSV+Al(OH)3+BCG组：有较明显的炎症细胞浸润，肺泡间隔增厚，肺泡融合，支气管周围存在炎症细胞浸润，但较FI-RSV+Al(OH)3组炎症减轻，与FI-RSV+BCG组相比，炎症症状较重。以上结果提示：在成年小鼠中，FI-RSV+BCG免疫接种方案可明显减轻肺部炎症病理。
实时定量PCR检测成年小鼠肺组织中RSV-N蛋白及细胞因子mRNA的表达：图4结果显示，与PBS/RSV组相比，FI-RSV+Al(OH)3、FI-RSV+BCG和FI-RSV+Al(OH)3+BCG组攻毒后的RSV-N基因(RSV病毒看家基因)的表达均显著减低，另外，FI-RSV+BCG和FI-RSV+Al(OH)3+BCG组的RSV-N的量显著低于FI-RSV+Al(OH)3组，表明FI-RSV+BCG和FI-RSV+Al(OH)3+BCG组提供了显著的保护性；
FI-RSV+BCG组的eotaxin(趋化嗜酸性粒细胞因子)或FI-RSV+Al(OH)3+BCG组的eotaxin(趋化嗜酸性粒细胞因子)、Gro-α(趋化中性粒细胞因子)、MCP-1(趋化单核细胞趋化蛋白1因子)与FI-RSV+Al(OH)3组相比明显降低；FI-RSV+BCG组的IL-17和IL-23(Th17型细胞因子)及FI-RSV+Al(OH)3+BCG组的IL-23也显著低于FI-RSV+Al(OH)3组；与FI-RSV+Al(OH)3组相比，FI-RSV+BCG和FI-RSV+Al(OH)3+BCG组的IL-13(Th2型细胞因子)显著减低，而IFN-γ(Th1型细胞因子)显著增高。
此部分实验结果提示：对于成年群体，FI-RSV+BCG为最佳免疫接种方案，既安全又有效。
以上所有实验结果表明：BCG作为FI-RSV(含或不含Al(OH)3)的佐剂均可明显改善FI-RSV或FI-RSV+Al(OH)3的保护性和安全性。
以上公开的本发明优选实施例只是用于帮助阐述本发明。优选实施例并没有详尽叙述所有的细节，也不限制该发明仅为所述的具体实施方式。显然，根据本说明书的内容，可作很多的修改和变化。本说明书选取并具体描述这些实施例，是为了更好地解释本发明的原理和实际应用，从而使所属技术领域技术人员能很好地理解和利用本发明。本发明仅受权利要求书及其全部范围和等效物的限制。
Claims (4)
1.卡介苗(BCG)作为制备治疗或预防呼吸道合胞病毒(Respiratory syncytialvirus，RSV)灭活疫苗佐剂的的用途，其特征在于：灭活疫苗为***灭活RSV疫苗FI-RSV。
2.如权利要求1所述的用途，其特征在于：所述灭活疫苗含或不含铝盐佐剂。
3.如权利要求1所述的用途，其特征在于：所述用途为将BCG提前或同时与RSV灭活疫苗注射体内，诱导平衡的免疫记忆。
4.如权利要求3所述的用途，其特征在于：所述用途为佐剂BCG仅注射一次或与灭活RSV疫苗注射相同次数。
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102307591A (zh) * | 2008-12-09 | 2012-01-04 | 诺瓦瓦克斯股份有限公司 | 修饰的rsv f蛋白及其使用方法 |
| CN104080476A (zh) * | 2011-09-30 | 2014-10-01 | 诺瓦瓦克斯股份有限公司 | 用于呼吸道合胞病毒的重组纳米颗粒rsv f疫苗 |
| WO2019191623A1 (en) * | 2018-03-30 | 2019-10-03 | Georgia State University Research Foundation, Inc. | Respiratory syncytial virus (rsv) vaccines |
-
2021
- 2021-07-30 CN CN202110873016.3A patent/CN113855796B/zh active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102307591A (zh) * | 2008-12-09 | 2012-01-04 | 诺瓦瓦克斯股份有限公司 | 修饰的rsv f蛋白及其使用方法 |
| CN104080476A (zh) * | 2011-09-30 | 2014-10-01 | 诺瓦瓦克斯股份有限公司 | 用于呼吸道合胞病毒的重组纳米颗粒rsv f疫苗 |
| JP2014530010A (ja) * | 2011-09-30 | 2014-11-17 | ノババックス，インコーポレイテッド | 呼吸器合胞体ウイルス用の組み換えナノ粒子ｒｓｖｆワクチン |
| CN105381457A (zh) * | 2011-09-30 | 2016-03-09 | 诺瓦瓦克斯股份有限公司 | 用于呼吸道合胞病毒的重组纳米颗粒rsv f疫苗 |
| WO2019191623A1 (en) * | 2018-03-30 | 2019-10-03 | Georgia State University Research Foundation, Inc. | Respiratory syncytial virus (rsv) vaccines |
| US20210052718A1 (en) * | 2018-03-30 | 2021-02-25 | Georgia State University Research Foundation, Inc. | Respiratory syncytial virus (rsv) vaccines |
Non-Patent Citations (8)
| Title |
|---|
| Jing Shan等.The immunogenicity and safety of respiratory syncytial virus vaccines in development: A systematic review.Influenza Other Respi Viruses.2021,第15卷(第4期),全文. * |
| Katherine M. Eichinger, Jessica L. Kosanovich, Madeline Lipp, Kerry M. Empey and Nikolai Petrovsky.Strategies for active and passive pediatric RSV immunization.Therapeutic Advances in Vaccines and Immunotherapy.2021,全文. * |
| Min Lin, Wei Zhang, Yi-Fan Yin, Jun-Yu Si, Lu-Jing Zhang, Li Chen, Xue Lin, Ying-Bin Wang, Jun Zhang,Zi-Zheng Zheng and Ning-Shao Xia.An Optimized FI-RSV Vaccine Effectively Protects Cotton Rats and BALB/c Mice without Causing Enhanced Respiratory Disease. Viruses.2022,第14卷(第10期),全文. * |
| Soto JA, Gálvez NMS, Rivera CA, et al. Recombinant BCG vaccines reduce pneumovirus-caused airway pathology by inducing protective humoral immunity.Front Immunol.2018,第9卷全文. * |
| Yang Wang, Fei Ge, Junhai Wang, Hanglin Li, Boyang Zheng, Wenjian Li, Shunyan Chen, Xiaoqing Zheng, Yuqing Deng, Yueling Wang, Ruihong Zeng.Mycobacterium bovis BCG Given at Birth Followed by Inactivated Respiratory Syncytial Virus Vaccine Prevents Vaccine-Enhanced Disease by Promoting Trained Macrophages and Resident Memory T Cells.J Virol.2023,第97卷(第3期),全文. * |
| 刘建东， 张静飞，刘建凯， 郑海发.人呼吸道合胞体病毒疫苗的研究进展. 中国生物制品学杂志.2021,第34卷(第2期),全文. * |
| 王洋，王俊海，陈顺妍，李文建，盖菲，李航林，曾瑞红.卡介苗和氢氧化铝双佐剂联合使用对RSV疫苗安全性和保护性的影响.第十四届全国免疫学学术大会论文摘要汇编.2021,全文. * |
| 王洋.卡介苗对FI-RSV疫苗安全性和保护性的调节作用及机制.河北医科大学硕士学位论文.2022,全文. * |
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