CN107375917A - 血吸虫病疫苗组合物及其使用方法 - Google Patents
血吸虫病疫苗组合物及其使用方法 Download PDFInfo
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Abstract
不存在对于毁灭性的血吸虫病有效的疫苗。本发明集中在Sm‑p80,其是在血吸虫免疫逃避过程中起到关键作用的、功能上重要的曼氏血吸虫抗原。当用于新的疫苗制剂中时,Sm‑p80显示了一致的免疫原性、保护潜能以及抗生殖效果。出于免疫的目的,制备了两种新型的DNA构建体。将Sm‑p80编码序列克隆到VR1020中。另外,将Sm‑p80编码序列克隆到在Sm‑p80序列的各个末端上具有侧翼的CpG基序的pcDNA3.1中。当用于不同的疫苗制剂中时两种构建体均显示了Sm‑p80的优越的抗生殖性和抗虫效果,其作为减少与血吸虫感染有关的发病率的重要疫苗候选者具有极大的潜力。
Description
本申请是2010年6月23日申请的PCT国际申请PCT/US2010/039707于2012年2月13日进入中国国家阶段的、申请号为201080035900.X且发明名称为“血吸虫病疫苗组合物及其使用方法”的发明专利申请的分案申请。
发明技术领域
本发明涉及针对曼氏血吸虫(Schistosoma mansoni)的DNA和蛋白质疫苗制剂。更具体地,本发明涉及用于控制人类寄生虫疾病-血吸虫病的组合物和方法。
发明背景
在不限制所公开的组合物和方法的范围的前提下,结合针对曼氏血吸虫(Schistosoma mansoni)的新的基于Sm-p80的疫苗制剂描述背景。
曼氏血吸虫(Schistosoma mansoni)是扁形虫寄生虫,其栖息于人类的门腔静脉-肠系膜循环中。据估计全世界有两亿零七百万人遭受几种血吸虫的折磨,导致相当高的发病率和致死率。还有七亿七千九百万人有遭受此感染的风险。血吸虫病的残疾调整生命年数(disability-adjusted life years)按照计算大约是一百七十万到四百五十万年之间。血吸虫病在74个发展中国家中流行。
曼氏血吸虫的感染周期涉及在蜗牛中间宿主中的无性繁殖,随后是对人类宿主的感染。从蜗牛中间宿主中离开的幼虫阶段,尾蚴通过穿透人类皮肤感染人类。这些幼年的血吸虫发育成熟成为童虫,经历通过宿主的肺和肝脏的复杂迁移,并且发育成为性成熟的产卵成虫。性成熟的雄性和雌性血吸虫在肠小静脉中开始生命周期的产卵阶段。大量卵的连续产生使得一些卵与粪便物质一起排出,以及在严重的感染中,卵在内脏器官中的停留导致了针对其的宿主肉芽瘤免疫应答。此卵诱导的器官损伤导致了如肝脏纤维化、门静脉高血压和食管静脉曲张等并发症,所述的并发症导致了慢性感染的宿主的死亡。
由于针对积累的含胚卵的肉芽瘤反应,这一使人虚弱的疾病的慢性性质导致了对肝脏、脾脏和结肠的累积性的损伤。感染导致了循环的抗血吸虫抗体的产生。免疫应答是无规律的,但其不产生无虫免疫。除此之外,成年的寄生虫通过复杂的以及多因子的机制逃避免疫清除。
在血吸虫病的治疗中,强调将化学疗法被作为优选方法。然而,基于化学疗法的控制方案由于快速和频繁的重复感染以及困难,以及长期维持这些项目所涉及的费用而复杂化。连续的药物治疗和重新感染周期不能充分地减少总体卵的产量,从而显著地降低该疾病在流行地区的传播。除此之外,有观点认为寄生虫可以产生抗药性。对于开发控制该疾病的替代方法还有着迫切的需要。
不存在对血吸虫病有效的疫苗。尽管可以使用抗寄生虫药物和其他的控制手段,包括公共保健和蜗牛控制,有效的疫苗的出现仍然是控制此疾病的潜在地最有效的方式。在幼龄时对个体进行接种是在不伴随着卵诱导的病症的情况下引发免疫系统的最有效方法。疫苗还能够预防严重的感染,并因此减少卵的传播,并且帮助抑制曼氏血吸虫感染的周期。可以在暴露于感染性的尾蚴后,对已接种的个体进行针对血吸虫的加强免疫接种。
有几种成虫的曼氏血吸虫蛋白质被认为是潜在的疫苗候选者。理想地,最为有希望的疫苗候选者可以是暴露在表面的,并且对于该寄生虫在人类宿主中的存活是必不可少的那些。
阻碍了血吸虫病疫苗研究和开发的主要问题涉及由寄生虫所编码的潜在的保护性抗原的鉴定和选择。在过去的二十年间,多个实验室尝试识别出诱导部分保护性免疫应答的血吸虫抗原。已经识别出了100个以上的这种抗原,其中约25%提供不同程度的保护。然而在这些候选者抗原中没有一个诱导的免疫应答的水平接近使用经辐照的血吸虫幼虫进行接种后所观察到的免疫水平(~80%)。通过标准比较性的世界卫生组织所描述的规程对六种“优先抗原”(副肌球蛋白、谷胱甘肽S-转移酶、14kDa的脂肪酸结合蛋白、IrV-5、丙糖磷酸异构酶和Sm23)进行的独立研究给出的结果是没有哪个抗原达到规定的40%或更高的保护的目标。
血吸虫与其宿主密切地相互作用,实现如免疫逃避、营养物摄取以及附着等功能。承担该关键功能的,暴露于宿主的血吸虫蛋白质是血吸虫病疫苗的有效靶。该蛋白的一种是钙蛋白酶的大亚基(Sm-p80),钙蛋白酶在血吸虫的表面膜更新中起到重要的作用,所述的表面膜更新是在血液寄生虫(blood-dwelling helminths)逃避宿主免疫所采取的免疫逃避机制。Sm-p80暴露在宿主和寄生虫的接触面上,并且是天然地免疫原性的。尽管此分子的天然免疫原性在自然感染的条件下不能提供保护,但有可能以诱导强有力的免疫力的方式将钙蛋白酶呈现到免疫系统。UNDP/World Bank/WHO-TDR专门小组将Sm-p80指定为“具有确定的背景,需要进一步开发”的优先抗原之一,并且现在本领域的国际专家认为Sm-p80是“一线候选者”之一。
有效的血吸虫病疫苗应该对当前的疾病控制规划带来突出的贡献,尤其是当其提供针对该疾病强有力的、长效的免疫力时。这样的疫苗能够极大地降低对于后勤困难以及昂贵的基于药物的方案的需求,所述的基于药物的方案通常需要政治上的保证和资金充足的公共健康系统才能进行。即使是针对尾蚴提供部分保护,也是显著的进步,因为减少虫负荷的疫苗能减轻疾病的病况并且能降低该疾病的传播速率。这是由于血吸虫不像其他的感染性有机体,不在其最终宿主中进行繁殖。因此,血吸虫病中可以不要求无虫免疫。世界卫生组织(WHO)的血吸虫病科学工作组确定,减少成虫虫负荷50%的疫苗在降低总体的发病率和死亡率中是有效的。
大多数血吸虫疫苗候选者在小鼠模型系统中产生30-50%的保护。因此,迫切需要识别新的抗原、佐剂载体和鸡尾酒疫苗制剂,从而诱导以辐照减毒的疫苗计70%到80%范围内的保护。
本发明提出了基于血吸虫蛋白质,钙蛋白酶的新的疫苗制剂,所述的钙蛋白酶最初确定为涉及血吸虫表面膜生物合成中。钙蛋白酶具有两个亚基,其中较大的亚基,Sm-p80显示出作为相关疫苗抗原的巨大潜力,所述的疫苗抗原用于降低与曼氏血吸虫及日本血吸虫(Schistosoma japonicum)有关的发病率。
发明简述
因此,本发明提供了用于控制人类寄生虫疾病,血吸虫病的组合物和方法。疫苗由Sm-p80,血吸虫蛋白质的各种制剂及递送方法组成。这是针对曼氏血吸虫(Schistosomamansoni)的首个有效的疫苗制剂。当前的控制策略仅得到有限的成功,所述的策略包括旨在通过改进教育和公共卫生来限制血吸虫病的整合的控制方案,用于减少蜗牛中间宿主的种群的软体动物杀灭剂治疗方案,以及化学疗法。因此,仍然迫切需要开发控制该疾病的替代方法,例如疫苗。
总而言之,本发明公开了用于新型的基于Sm-p80的针对曼氏血吸虫(S.mansoni)的DNA疫苗制剂的组合物和方法。
附图简要说明
为了更完整地理解本发明的特征和优点,在此参考本发明的详述以及其所附的图,其中:
图1是对DNA构建体的描述,其中将Sm-p80编码序列克隆到VR1020中,从而成为根据本发明公开内容的实施方案的DNA疫苗制剂之一;
图2是对DNA构建体的描述,其中将Sm-p80编码序列克隆到pcDNA 3.1中,在Sm-p80序列的各个末端上具有侧翼的CpG基序,从而构成了根据本发明公开内容的实施方案的另一种DNA疫苗制剂;
图3是对构建根据本发明公开内容的实施方案的VR1020/Sm-p80和pcDNA3/Sm-p80的第一种方法的描述;
图4是对构建根据本发明公开内容的实施方案的pcDNA3/Sm-p80和VR1012/Sm-p80的第二种方法的描述;
图5是对用对照质粒VR1020(n=10)和用Sm-p80-VR1020(n=10)免疫的各组小鼠虫负荷分布的描述。在经接种的动物中,虫负荷的减少在统计上更低(P<0.001);
图6是对在经免疫的小鼠中的抗-Sm-p80总IgG的抗体滴度的描述。将收集自各自的组(VR1020和Sm-p80-VR1020)中的各个小鼠(每两周地)的等体积的血清混合得到血清池,用所述的血清池进行ELISA。数值代表了三次试验的平均值±标准偏差。与VR1020组进行比较的统计上的显著性(P≤0.05)通过(*)表示;
图7是对在经免疫的小鼠中的抗-Sm-p80总IgM的抗体滴度的描述。将收集自各自的组(VR1020和Sm-p80-VR1020)中的各个小鼠(每两周地)的等体积的血清混合得到血清池,用所述的血清池进行ELISA。数值代表了三次试验的平均值±标准偏差。与VR1020组进行比较的统计上的显著性(P≤0.05)通过(*)表示;
图8是对在经免疫的小鼠中的抗-Sm-p80IgG2a的抗体滴度的描述。将收集自各自的组(VR1020和Sm-p80-VR1020)中的各个小鼠(每两周地)的等体积的血清混合得到血清池,用所述的血清池进行ELISA。数值代表了三次试验的平均值±标准偏差。与VR1020组进行比较的统计上的显著性(P≤0.05)通过(*)表示;
图9是对在经免疫的小鼠中的抗-Sm-p80IgG2b的抗体滴度的描述。将收集自各自的组(VR1020和Sm-p80-VR1020)中的各个小鼠(每两周地)的等体积的血清混合得到血清池,用所述的血清池进行ELISA。数值代表了三次试验的平均值±标准偏差。与VR1020组进行比较的统计上的显著性(P≤0.05)通过(*)表示;
图10描述了在体外培养48小时之后,与伴刀豆球蛋白A(Concanavalin A)所诱导的刺激相比,重组的Sm-p80诱导的脾细胞增殖;
图11描述了在体外培养48小时之后,重组的Sm-p80诱导的脾细胞增殖;
图12描述了在使用重组的Sm-p80在体外刺激48小时之后脾细胞产生的细胞因子水平。各组小鼠使用VR1020和VR1020-Sm-p80进行接种。数据表示为平均值±标准偏差。使用独立样本检验与VR1020组进行比较的统计上的显著性(P≤0.05)通过(*)表示;
图13A描述了RT-PCR之后的琼脂糖凝胶(白细胞介素6)。M=100bp标记物;1=IL-6(VR1020组);2=IL-6(VR1020-Sm-p80组);
图13B描述了RT-PCR之后的琼脂糖凝胶(白细胞介素6)。M=100bp标记物;1=IL-6(VR1020组);2=IL-6(VR1020-Sm-p80组);
图13C描述了RT-PCR之后的琼脂糖凝胶(白细胞介素6)。M=100bp标记物;1=IL-6(VR1020组);2=IL-6(VR1020-Sm-p80组);
图14A描述了RT-PCR之后的琼脂糖凝胶(甘油醛-3-磷酸脱氢酶)。M=100bp标记物;1=GAPDH(VR1020组);2=GAPDH(VR1020-Sm-p80组);
图14B描述了RT-PCR之后的琼脂糖凝胶(甘油醛-3-磷酸脱氢酶)。M=100bp标记物;1=GAPDH(VR1020组);2=GAPDH(VR1020-Sm-p80组);
图15描述了RT-PCR之后的琼脂糖凝胶(甘油醛-3-磷酸脱氢酶和白细胞介素1α)。M=100bp标记物;1=GAPDH(VR1020组);2=GAPDH(VR1020-Sm-p80组);3=IL-1α(VR1020组);4=IL-1α(VR1020-Sm-p80组);
图16描述了RT-PCR之后的琼脂糖凝胶(白细胞介素1α)。M=100bp标记物;1=IL-1α(VR1020组);2=IL-1α(VR1020-Sm-p80组);
图17描述了RT-PCR之后的琼脂糖凝胶(干扰素γ)。M=100bp标记物;1=IFN-γ(VR1020组);2=IFN-γ(VR1020-Sm-p80组);
图18描述了RT-PCR之后的琼脂糖凝胶(白细胞介素4)。M=100bp标记物;1=IL-4(VR1020组);2=IL-4(VR1020-Sm-p80组);
图19描述了RT-PCR之后的琼脂糖凝胶(白细胞介素5)。M=100bp标记物;1=IL-5(VR1020组);2=IL-5(VR1020-Sm-p80组);
图20描述了RT-PCR之后的琼脂糖凝胶(白细胞介素17)。M=100bp标记物;1=IL-17(VR1020组);2=IL-17(VR1020-Sm-p80组);
图21描述了RT-PCR之后的琼脂糖凝胶(白细胞介素2)。M=100bp标记物;2=IL-2(VR1020-Sm-p80组);1=IL-2(VR1020组);
图22描述了RT-PCR之后的琼脂糖凝胶(肿瘤坏死因子α)。M=100bp标记物;1=TNF-α(VR1020组);2=TNF-α(VR1020-Sm-p80组);
图23描述了RT-PCR之后的琼脂糖凝胶(白细胞介素1β)。M=100bp标记物;1=IL-1β(VR1020组);2=IL-1β(VR1020-Sm-p80组);
图24描述了在VR1020组和VR1020-Sm-p80组中,RT-PCR之后的琼脂糖凝胶(GAPDH、IL-1α、IL-1β、IL-2、IL-4、IL-5、IL-6、IL-17和TNF-α);
图25描述了对于使用对照质粒VR1020(n=6)和使用VR1020-Sm-p80(n=6)进行免疫的狒狒的组中,在狒狒的个体中每克的肝脏和肠中的卵的负载。在经接种的动物中,卵数量的减少统计上较低(P<0.05);
图26描述了对于使用对照质粒VR1020(n=6)和使用VR1020-Sm-p80(n=6)进行免疫的狒狒的组中虫负荷分布。在经接种的动物中,虫负荷的减少统计上较低(P<0.05);
发明详述
在此公开了用于针对曼氏血吸虫的基于Sm-p80的疫苗制剂的组合物和方法。本发明的多种创新性的教导将具体参考几个实施方案(其以举例而非限制的形式)进行描述。
首先参考图1,VR1020/Sm-p80构建体的示意图。图1,两种构建体的第一种,描绘了通过将血吸虫Sm-p80编码序列克隆到VR1020中所产生的DNA构建体。
现参考图2,pcDNA3.1/Sm-p80构建体的示意图。图2,两种构建体的第二种,描绘了通过将血吸虫Sm-p80编码序列克隆到pcDNA 3.1中所产生的DNA构建体,所述的DNA构建体在Sm-p80序列的各个末端上具有侧翼的CpG基序。本领域普通技术人员能依赖图2构建该DNA疫苗。
现参考图3,举例说明了构建VR1020/Sm-p80和pcDNA3/Sm-p80的第一种方法的示意图。
现参考图4,举例说明了构建VR1020/Sm-p80和pcDNA3/Sm-p80的第二种方法的示意图。
本领域普通技术人员能够依靠图1与图3或者图4的结合,构建DNA疫苗。
在图1和图2中描述的两种构建体都用于不同的疫苗制剂中(单独的DNA和初免加强(prime boost)),所述的疫苗制剂递送到小鼠和狒狒中。这是血吸虫病的狒狒模型中基于Sm-p80的疫苗制剂的首次使用。表1总结了1.基于Sm-p80的疫苗制剂[(a)单独的DNA疫苗,(b)在构建体中插入了两个未甲基化的CpG基序的DNA疫苗,由于其作为免疫刺激剂起作用,(c)重组的Sm-p80蛋白质在含有CpG基序的寡脱氧核苷酸(ODN)的存在下引入,激活宿主的防御机制,导致先天性的和获得性的免疫应答)2.疫苗递送途径和3.所得到的结果。在鼠科动物以及非人类的灵长类动物模型中,Sm-p80抗生殖和抗虫效果的实验数据清楚地表明此抗原作为用于减少与血吸虫感染相关的发病率的重要疫苗候选者,具有重大的潜力。总之,基于Sm-80的疫苗制剂具有三种保护性的效果(减少虫,抗生殖效果和针对急性血吸虫病进行保护)。这是显示了这三种保护性效果的、对抗血吸虫的经确定的疫苗制剂的首次报道。
表2和表3详述了用于将小鼠使用基于pcDNA的载体进行免疫的方案。
表4和表5详述了用于将小鼠使用基于VR1020的载体进行免疫的方案。
用于狒狒的免疫的方案包括裸露DNA接种,以及首次加强和蛋白质接种策略。在狒狒中使用裸露DNA接种的方案如下所示:
第1组:对于此对照组而言,初次的免疫使用500或者1000μg质粒DNA(没有插入物)。将该DNA经肌肉内(IM)注射于四头肌中。在第4、8和12周使用500或者1000μg的对照质粒DNA对狒狒进行加强。在全体的接种中,使用500μg,由于在许多非人类的灵长类动物系统中,这一量提供一致的结果。
第2组:为了测定单独的Sm-p80的保护效果,最初的免疫采用500μg质粒Sm-p80-pcDNA3.1或者Sm-p80-VR1020完成。将该DNA IM注射于四头肌中。在第4、8和12周用500μgSm-p80-pcDNA3.1或者Sm-p80-VR1020对狒狒进行加强。
第3组:为了确定使用IL-2作为遗传佐剂时Sm-p80的保护效果是否可以增强,采用500μg质粒Sm-p80-pcDNA3或Sm-p80-VR1020与500μg质粒pORF-hIL-2进行最初的免疫。将该DNA IM注射于四头肌中。在第4、8和12周用500μg Sm-p80-pcDNA3.1或者Sm-p80-VR1020与500μg质粒pORF-hIL-2对狒狒进行加强。
在最后一次加强4周后,通过腹袋法(abdominal pouch method),将来自所有组的狒狒使用总计1000只曼氏血吸虫的尾蚴进行激发。在初次激发后8周,将狒狒进行固定,并且使用氯胺酮(Ketaminol-10mg/kg体重)和甲苯噻嗪(0.5mg/kg)的混合物进行轻度麻醉,然后通过静脉注射肝素化的戊巴比妥钠溶液进行深度麻醉。随后通过从心室放血安乐处死该动物。此安乐处死的方法通过肠系膜静脉的灌注及灌注后的检查,有助于成虫的定量回收。通过改进已公开的方法,成虫寄生虫通过从肠系膜脉管系统和肝门系统的灌注进行回收。保护(P)按照将虫负荷在接种的(V)和对照的(C)狒狒之间,通过标准式:%P=(C-V)/(Cx 100)进行比较计算得到。
在狒狒中使用初次加强和蛋白质接种策略的方案如下所示:
在小鼠中提供最优保护结果的初免/加强方法已经在狒狒中使用。将动物使用500μg质粒DNA(Sm-p80-VR1020或者免疫刺激序列(ISS)-Sm-p80-ISS-VR1020)进行初次免疫,并使用200μg杆状病毒产生的重组Sm-p80蛋白在ODN#2138(250μg)或者瑞喹莫德(50μg)的存在下进行加强。在年龄匹配的对照组中的动物接受与实验组中动物类似的量的无插入物的质粒DNA,并且使用与实验组中相同的佐剂以不相关的由BEVS产生的重组蛋白进行加强。还包括单独的抗原DNA(没有加强)以及单独的重组蛋白(没有初免)作为对照。注意到,用于产生所述的Sm-p80-ISS构建体的ISS序列以及本研究中所使用的CpG ODN一致地显示在小鼠及非人类的灵长类动物中都有效地起作用。
现参考图5-图24,其与表6-表24一起详述了得自在小鼠和狒狒的体外试验及体内试验的实验结果。这些实验结果证实了本发明的效果。
现参考图5并结合表6,证实了在用对照质粒VR1020(n=10)和使用Sm-p80-VR1020(n=10)进行免疫的小鼠的组中虫负荷分布的减少。使用Sm-p80-VR1020进行免疫的小鼠与仅接受对照质粒VR1020的小鼠相比,显示出虫负荷减少46.87%。虫负荷的减少在接种的动物中统计上是显著的(P<0.001)。
现参考图6,在免疫的小鼠中抗-Sm-p80总IgG的抗体滴度的图。表7列出了接种重组Sm-p80疫苗所诱导的血清抗体总IgG产量。
现参考图7,其是在免疫的小鼠中,抗-Sm-p80总IgM的抗体滴度的图。表8列出了接种重组Sm-p80疫苗诱导的血清抗体IgM产量。
现参考图8,其是在免疫的小鼠中,抗-Sm-p80IgG2a的抗体滴度的图。表9列出了接种重组Sm-p80疫苗诱导的血清抗体IgG2a产量。
现参考图9,其是在免疫的小鼠中,抗-Sm-p80IgG2b的抗体滴度的图。表10列出了接种重组Sm-p80疫苗诱导的血清抗体IgG2b产量。
现参考图10和图11并结合表11,其证实了在体外培养48小时之后,与由伴刀豆球蛋白A诱导的刺激相比,重组的Sm-p80诱导的脾细胞增殖。
现参考图12,其描述了使用重组Sm-p80在体外进行刺激48小时之后,脾细胞产生的细胞因子的水平(也见于表12)。各组小鼠使用VR1020和VR1020-Sm-p80进行接种。数据显示为平均值±标准偏差。使用独立样本检验与VR1020组进行比较的统计上的显著性(P≤0.05)通过(*)表示。
现参考图13-图24,其描述了在VR1020组和VR1020-Sm-p80组的免疫的小鼠中,所评价的各种细胞因子(GAPDH、IL-1α、IL-1β、IL-2、IL-4、IL-5、IL-6、IL-17和TNF-α)的琼脂糖凝胶电泳。表13定量地分析在VR1020组和VR1020-Sm-p80组的免疫的小鼠中评估的各种细胞因子。
表14总结了在狒狒用VR1020接种的对照组中血清抗体滴度的产量,以及在狒狒用VR1020-Sm-p80接种的试验组中血清抗体滴度的产量
表15和表16总结了在体外使用重组Sm-p80刺激48小时后,脾细胞产生的细胞因子的水平。将各组狒狒使用VR1020(对照组)和VR1020-Sm-p80(试验组)进行接种。各组数据表示为平均值±标准偏差。
表17和表18总结了使用重组Sm-p80在体外刺激48小时之后,外周血液单核细胞所产生的细胞因子的水平。狒狒的组使用VR1020(对照组)和VR1020-Sm-p80(试验组)进行接种。数据表示为平均值±标准偏差。
表19和表20总结了在体外培养48小时之后,重组Sm-p80诱导的白细胞介素4和干扰素γ的点形成单位(SFU)。各个狒狒使用VR1020(对照组)或者VR1020-Sm-p80(试验组)进行接种。数据表示为平均值±标准偏差。
一般性地描述了所公开的疫苗组合物和使用的方法,引入了作为本发明具体实施方案的实施例以证实其实践和优点。应理解,所述的实施例以举例说明的方式给出,而不意在以任何方式限制说明书或者权利要求书。
为了便于理解本发明,在下文中定义了多个术语。在此定义的术语具有本发明相关的领域内普通技术人员一般所理解的含义。如“一个(a)”、“一个(an)”和“该(the)”不期待仅指单个的实体,而是包括一般的类,所述的类中特定的实例可以用于举例说明。在此的术语用于描述本发明的特定的实施方案,但是其使用不限制所公开的方法,除非在权利要求书中进行概述。
应当理解在此描述的特定实施方案是以举例说明的方式显示的,而非作为本发明的限制。本发明的主要特征可以在不同的实施方案中使用而不背离本发明的范围。本领域的技术人员能够认识到,或者能够使用不多于常规的试验确定与在此所描述的特定的过程和疫苗组合物的多种等价物。该等价物认为是在本发明的范围内的,并且由权利要求书所涵盖。
在本说明书中所提及的所有出版物和专利申请表明了本发明所属领域技术人员的水平。在此将所有的出版物和专利申请引入作为参考,其参考的程度如同各个单独的出版物或专利申请具体地和单独地引入作为参考的程度相同。
在权利要求书中,过渡短语如“包含”、“包括(including)”、“带有”、“具有”、“含有”、“包括(involving)”等等理解为开放式的,也即其表示包括但是不限于。仅有过渡短语“由…组成”以及“基本上由…组成”分别地应是封闭的或者半封闭的过渡短语。
按照本公开内容,在此公开和要求保护的所有疫苗组合物和/或使用的方法可以在不过度的试验下实施和实现。尽管本发明的疫苗组合物和使用的方法是以优选的实施方案的形式进行描述的,对于本领域技术人员而言显而易见的是,在不背离本发明的构思、精神和范围的前提下,可以在疫苗组合物和/或使用的方法,以及在此描述的方法的步骤或者步骤的顺序中进行改变。
更具体地,明显的是只要可以实现同样的或者类似的结果,在材料和功能上相关的组件可以取代为在此描述的组件。所有的该类似的取代和改变对于本领域技术人员来说是显而易见的,并认为在如所附的权利要求书所定义的本发明的精神、范围和构思之内。
参考文献
1.Gryseels B,Polman K,Clerinx J,Kestens L.Humanschistosomiasis.Lancet 2006;368(September(9541)):1106-18.
2.Steinmann P,Keiser J,Bos R,Tanner M,Utzinger J.Schistosomiasis andwater resources development:systematic review,meta-analysis,and estimates ofpeople at risk.Lancet Infect Dis 2006;6(July(7)):411-25.
3.Bergquist R,Al-Sherbiny M,Barakat R,Olds R.Blueprint forschistosomiasis vaccine development.Acta Trop 2002;82(May(2)):183-92.
4.Siddiqui AA,Ahmad G,Damian RT,Kennedy RC.Experimental vaccines inanimal models for schistosomiasis.Parasitol Res 2008;102(April(5)):825-33.
5.McManus DP,Loukas A.Current status of vaccines forschistosomiasis.Clin Microbiol Rev 2008;21(January(1)):225-42.
6.Siddiqui AA,Zhou Y,Podesta RB,Karcz SR,Tognon CE,Strejan GH,etal.Characterization of Ca(2+)-dependent neutral protease(calpain)from humanblood flukes,Schistosoma mansoni.Biochim Biophys Acta 1993;1181(March(1)):37-44.
7.Karcz SR,Podesta RB,Siddiqui AA,Dekaban GA,Strejan GH,ClarkeMW.Molecular cloning and sequence analysis of a calcium-activatedneutral protease(calpain)from Schistosoma mansoni.Mol Biochem Parasitol 1991;49(December(2)):333-6.
8.Silva EE,Clarke MW,Podesta RB.Characterization of a C3receptor onthe envelope of Schistosoma mansoni.J Immunol 1993;151(December(12)):7057-66.
9.Young BW,Podesta RB.Complement and 5-HT increasephosphatidylcholine incorporation into the outer bilayers ofSchistosomamansoni.J Parasitol 1986;72(October(5)):802-3.
10.Van Hellemond JJ,Retra K,Brouwers JF,et al.Functions of thetegument of schistosomes:clues from the proteome and lipidome.Int J Parasitol2006;36(May (6)):691-9.
11.Ahmad G,Torben W,Zhang W,Wyatt M,Siddiqui AA.Sm-pS0based DNAvaccine formulation induces potent protective immunity against Schistosomamansoni.Parasite Immunol 2009;31(March(3)):156-61.
12.Hota-Mitchell S,Siddiqui AA,Dekaban GA,Smith J,Tognon C,PodestaRB.Protection against Schistosoma mansoni infection with a recombinantbaculovirus-expressed subunit of calpain.Vaccine 1997;15(October(15)):1631-40.
13.Hota-Mitchell S,Clarke MW,Podesta RB,Dekaban GA.Recombinantvaccinia viruses and gene gun vectors expressing the large subunitofSchistosoma mansoni calpain used in a murine immunization-challengemodel.Vaccine 1999;17(March(11-12)):1338-54.
14.Siddiqui AA,Phillips T,Charest H,Podesta KB,Quinlin ML,PinkstonJR,et al.Enhancement of Sm-p80(large subunit of calpain)induced protectiveimmunity against Schistosoma mansoni through co-delivery of interieukin-2 andinterleukin-12 in a DNA vaccine formulation.Vaccine 2003;21(June(21-22)):2882-9.
15.SiddiquiAA,Pinkston JR,Quinlin ML,Kavikondala V,Rewers-FelkinsKA,Phillips T,et al.Characterization of protective immunity induced againstSchistosoma mansoni via DNA priming with the large subunit of calpain(Sm-p80)in the presence of genetic adjuvants.Parasite 2005;12(March(1)):3-8.
16.Jankovic D,Aslund L,Oswald IP,Caspar P,Champion C,Pearce E,etal.Calpain is the target antigen of a Th1 clone that transfers protectiveimmunity against Schistosoma mansoni.J Immunol 1996;157(July(2)):806-14.
17.Ohta N,Kumagai T,Maruyama H,Yoshida A,He Y,Zhang R.Research oncalpain of Schistosoma japonicum as a vaccine candidate.Parasitol Int 2004;53(June(2)):175-81.
18.Ridi RE,Tallima H.Schistosoma mansoni ex vivo lung-stage larvaeexcretory-secretory antigens as vaccine candidates againstschistosomiasis.Vaccine 2009;27(5):666-73.
19.Zhang R,Yoshida A,Kumagai T,Kawaguchi H,Maruyama H,Suzuki T,etal.Vaccination with calpain induces a Th1-biased protective immune responseagainst Schistosoma japonicun.Infect Immun 2001;69(January(1)):386-91.
20.Kennedy RC,Shearer MH,Hildebrand W.Nonhuman primate models toevaluate vaccine safety and immunogenicity.Vaccine 1997;15(June(8)):903-8.
21.Siddiqui AA,Phillips T,Charest H,Podesta RB,Quinlin ML,PinkstonJR,et al.Induction of protective immunity against Schistosoma mansoni via DNApriming and boosting with the large subunit of calpain(Sm-p80):adjuvanteffects of granulocyte-macrophage colony-stimulating factor and interleukin-4.Infect Immun 2003;71(July (7)):3844-51.
22.Siddiqui AA,Pinkston JR,Quinlin ML,Saeed Q,White GL,Shearer MH,etal.Characterization of the immune response to DNA vaccination strategies forschistosomiasis candidate antigen,Sm-p80 in the baboon.Vaccine 2005;23(February (12)):1451-6.
23.Smithers SR,Terry RJ.The infection of laboratory hosts withcercariae of Schistosoma mansoni and the recovery of the adultworms.Parasitology 1965;55(November(4)):695-700.
24.Damian RT,Greene ND,Fitzgerald K.Schistosomiasis mansoni inbaboons.The effect of surgical transfer of adult Schistosoma mansoni uponsubsequent challenge infection.Am J Trop Med Hyg 1972;21(November(6)):951-8.
25.Cheever AW.Conditions affecting the accuracy of potassiumhydroxide digestion techniques for counting Schistosoma mansoni eggs intissues.Bull World Health Organ 1968;39(2):328-31.
26.Shearer MH,Dark RD,Chodosh J,Kennedy RC.Comparison andcharacterization of immunoglobulin G subclasses among primate species.ClinDiagn Lab Immunol 1999;6(November(6)):953-8.
27.Vereecken K,Naus CW,Polman K,Scott JT,Diop M,Gryseels B,etal.Associations between specific antibody responses and resistance toreinfection in a Senegalese population recently exposed to Schistosomamansoni.TropMed Iht Health 2007;12(March(3)):431-44.
28.Acosta LP,Waine G,Aligui GD,Tiu WU,Olveda RM,McManus DP.Immunecorrelate study on human Schistosoma japonicum in a well-defined populationin Leyte,Philippines.II.Cellular immune responses to S.japonicum recombinantand native antigens.Acta Trop 2002;84(November(2)):137-49.
29.Olds GR.New insights into the observed age-specific resistance toreinfection with Schistosoma japonicum.Clin Infect Dis 2006;42(June(12)):1699-701.
30.Hewitson JP,Hamblin PA,Mountford AP.Immunity induced by theradiation-attenuated schistosome vaccine.Parasite Immunol 2005;27(July(7-8)):271-80.
31.Lightowlers MW.Cestode vaccines:origins,current status and futureprospects.Parasitology 2006;133(Suppl.):S27-42.
32.Vercruysse J,Schetters TP,Knox DP,Willadsen P,Claerebout E.Controlof parasitic disease using vaccines:an answer to drug resistance?Rev Sci Tech2007;26(April (1)):105-15.
33.Kumagai T,Maruyama H,Hato M,Ohmae H,Osada Y,Kanazawa T,etal.Schistosoma japonicum:localization of calpain in the penetration glandsand secretions of cercariae.Exp Parasitol 2005;109(January(1)):53-7.
34.Damian RT,de la Rosa MA,Murfin DJ,Rawlings CA,Weina PJ,XueYP.Further development of the baboon as a model for acute schistosomiasis,MemInst Oswaldo Cruz 1992;87(Suppl.4):261-9.
35.Nyindo M,Farah IO.The baboon as a non-human primate model of humanschistosome infection.Parasitol Today 1999;15(December(12)):478-82.
36.Boulanger D,Reid GD,Sturrock RF,Wolowczuk I,Balloul JM,Grezel D,etal.Immunization of mice and baboons with the recombinant Sm28GST affects bothworm viability and fecundity after experimental infection with Schistosomamansoni.Parasite Immunol 1991;13(September(5)):473-90.
37.Kanamura HY,Hancock K,Rodrigues V,Damian RT.Schistosoma mansoniheat shock protein 70 elicits an early humoral immune response in S.mansoniinfected baboons.Mem Inst Oswaldo Cruz 2002;97(July(5)):711-6.
38.Kariuki TM,Farah IO,Yole DS,Mwenda JM,Van Dam GJ,Deelder AM,etal.Parameters of the attenuated schistosome vaccine evaluated in the olivebaboon.Infect Immun 2004;72(September(9)):5526-9.
39.Reid GD,Sturrock RF,Harrison RA,Tarara RP.Schistosoma haematobiumin the baboon(Papio anubis):assessment of protection levels against either asingle mass challenge or repeated trickle challenges after vaccination withirradiated schistosomula.J Helminthol 1995;69(June(2)):139-47.
40.Soisson LA,Reid GD,Farah IO,Nyindo M,Strand M.Protective immunityin baboons vaccinated with a recombinant antigen or radiationattenuatedcercariae of Schistosomamansoni is antibody-dependcnt.J Immunol 1993;151(November(9)):4782-9.
41.Yole DS,Pemberton R,Reid GD,Wilson RA.Protective immunity toSchistosoma mansoni induced in the olive baboon Papio anubis by theirradiated cercaria vaccine.Parasitology 1996;112(January(Pt 1)):37-46.
42.Kariuki TM,Farah IO.Resistance to re-infection after exposure tonormal and attenuated schistosome parasites in the baboon model.Parasitelmmunol 2005;27(July(7-8)):281-8.
43.Stacy S,Pasquali A,Sexton VL,Cantwell AM,Kraig E,Dube PH.An ageoldparadigm challenged:old baboons generate vigorous humoral immune responses toLcrV,a plague antigen.J Immunol 2008;181(July(1)):109-15.
44.Coulson PS,Kariuki TM.Sehistosome vaccine testing:lessons from thebaboon model.Mem Inst Oswaldo Cruz 2006;101(September(Suppl.1)):369-72.
45.Wilson RA,Langermans JA,van Dam GJ,Vervenne RA,Hall SL,Borges WC,et al.Elimination of Schistosoma mansoni adult worms by rhesus macaques:basisfor a therapeutic vaccine?PLoS Negl Trop Dis 2008;2(9):e290.
46.Ahmad G,Zhang W,Torben W,Damian RT,WolfRF,White GL,Chavez-SuarezM,Kennedy RC,Siddiqui AA.Protective and antifecundity effects of Sm-p80-basedDNA vaccine formulation against Schistosoma mansoni in a nonhuman primatemodel.Vaccine 27(2009):2830-2837.
Claims (12)
1.一种用于预防血吸虫病的方法,所述方法包括以下步骤:
给予有效剂量的疫苗,所述疫苗包含单一表达载体,所述单一表达载体包含:
pcDNA3.1表达载体;
曼氏血吸虫钙蛋白酶(Sm-p80)的大亚基的全长cDNA;和
作为佐剂的侧翼CpG寡核苷酸,
其中有效剂量是足以在宿主中实现虫减少、抗生殖力效果或者针对急性血吸虫病的保护的量。
2.权利要求1所述的方法,其中所述载体是VR1020。
3.权利要求1所述的方法,其中在第0周给予所述疫苗进行初免,在第4周进行第一次加强,并且在第8周进行第二次加强。
4.权利要求1所述的方法,其中在第0周给予所述疫苗进行初免,还包括以下步骤:在第4周给予包含所述疫苗和作为佐剂的Th1应答增强剂的第一次加强,并且在第8周给予包含所述疫苗和作为佐剂的Th1应答增强剂的第二次加强。
5.权利要求1所述的方法,其还包括Th1应答增强剂佐剂。
6.权利要求5所述的方法,其中所述Th1应答增强剂佐剂包括一种或多种CpG寡核苷酸。
7.权利要求5所述的方法,其中所述佐剂是免疫调节剂瑞喹莫德(R848)。
8.一种血吸虫病疫苗,其包含:
克隆到pcDNA3.1表达载体中的曼氏血吸虫钙蛋白酶(Sm-p80)的大亚基的全长cDNA和作为佐剂的侧翼CpG寡核苷酸。
9.权利要求8所述的疫苗,其中所述载体是VR1020。
10.权利要求8所述的疫苗,其还包含Th1应答增强剂佐剂。
11.权利要求8所述的疫苗,其中所述Th1应答增强剂佐剂包括CpG寡核苷酸。
12.权利要求10所述的疫苗,其中所述佐剂是免疫调节剂瑞喹莫德(R848)。
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