CN1329513C - 抗猪生殖和呼吸疾病的多核苷酸疫苗制剂 - Google Patents
抗猪生殖和呼吸疾病的多核苷酸疫苗制剂 Download PDFInfo
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- CN1329513C CN1329513C CNB971965587A CN97196558A CN1329513C CN 1329513 C CN1329513 C CN 1329513C CN B971965587 A CNB971965587 A CN B971965587A CN 97196558 A CN97196558 A CN 97196558A CN 1329513 C CN1329513 C CN 1329513C
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Abstract
抗猪生殖和/或呼吸疾病的猪疫苗制剂,所述制剂含有至少3种效价的多核苷酸疫苗,含有一个整合质粒,其可在宿主细胞中体内表达一种猪病原体效价的基因。这些效价选自两组,即Aujeszkey病病毒、猪流感病毒、猪神秘病病毒、细小病毒病病毒、典型猪瘟病毒和引起放线杆菌病的细菌。对于每种效价而言,质粒含有一种或多种基因,该基因对于Aujeszkey病病毒选自gB和gD,对于猪流感病毒选自HA,NP和N,对于神秘病病毒选自E、ORF3、M,对于细小病毒病病毒选自VP2,对于瘟病毒病病毒选自E1、E2,对放线杆菌病选自apxⅠ、apxⅡ和apxⅢ。
Description
本发明涉及可特别地接种猪以抗生殖和呼吸疾病的疫苗制剂,还涉及相应的接种方法。
在过去几十年里,生产猪的方法发生了根本性的变化。在被包围的空间内集中繁殖已被推广,但其必然后果是呼吸疾病也随之迅速发展。
猪呼吸病理学的症状范围一般被归纳为复杂的猪呼吸疾病名称,并涉及多种致病因子,其中包括病毒以及细菌和支原体。
参与呼吸疾病的主要因子是大叶性肺炎放线杆菌(Actino-bacilluspleuropneumoniae),不育和呼吸综合征病毒(PRRS),也被称为神秘病病毒,Aujeszkey病病毒(PRV)和猪流感病毒。
其它病毒导致生殖疾病,所述疾病导致流产,胎儿干瘪和不育。主要病毒是PRRS,细小病毒和典型猪瘟病毒(HCV)。其次,猪流感病毒PRV和大叶性肺炎放线杆菌也可导致这种疾病。大叶性肺炎放线杆菌、HCV和PRV可导致死亡。
另外,微生物之间的相互关系在猪呼吸综合征中非常重要。实际上,大多数细菌病原体是鼻咽区和幼小动物扁桃体的习惯性宿主。来自大母猪的这些病原体经常被出生仅几个小时、初乳免疫力尚未起效之前的小猪吸入。当宿主的呼吸防御机制被诸如大叶性肺炎放线杆菌的前体因子或病毒损坏时,在上呼吸道中生存的生物体可入侵下呼吸道。对肺的入侵非常快,尤其是在诸如大叶性肺炎放线杆菌的前体病原体的情况下更是如此,所述病原体可产生强有力的细胞毒素,该毒素能损害呼吸道上皮细胞的纤毛和肺泡巨噬细胞。
除了具有呼吸道趋向性的细菌和支原体外,主要的病毒感染,如流感病毒,呼吸道冠状病毒和Aujeszkey病病毒的感染对呼吸道综合征的致病性可能也起到了一定的作用。
最后,一些因子既有呼吸效应也有生殖效应。从生殖病理学的角度看,也会出现相互作用。
因此,似乎需要试着开展针对猪生殖和呼吸疾病的主要致病因子的有效防治作用。
迄今为止发展起来的方法是用灭活疫苗或活疫苗,及可能为这种疫苗的混合物。它们的发展提出了效价和稳定性之间相容性的问题。实际上不论是从所用的不同抗原的角度,还是从制剂本身的角度,尤其是在联合使用灭活疫苗和活疫苗的情况下,都必需确保不同疫苗效价之间的相容性。另外还存在的问题是:这种联合疫苗的保存及其安全性,尤其是存在佐剂时的安全性。这些疫苗一般十分昂贵。
专利申请WO-A-90 11092,WO-A-92 19183,WO-A-94 21797和WO-A-95 20660利用了最近兴起的多核苷酸疫苗技术。已知这些疫苗使用了质粒,所述质粒能在宿主细胞中表达插入其中的抗原。已提出所有的施用途径(腹膜内,静脉内,肌内,经皮,皮内,粘膜等等)。也可以使用不同的接种方式,如将DNA沉积在金粒表面并投射之以穿入动物皮肤(Tang等,自然(Nature),356,152-154,1992),可以同时转染皮肤,肌肉,脂肪组织和乳房组织的液体喷射注射器(Furth等,分析生物化学(Analytical Biochemistry),205,365-368,1992)。
多核苷酸疫苗也可以使用裸DNA和经配制的DNA,如阳离子脂质脂质体内的DNA。
M-F Le Potier等人(第二届消灭Aujeszkey病(假狂犬病)病毒国际研讨会论文集,1995年8月6日至8日,哥本哈根,丹麦)和M.Monteil等人(Les Journees d’Animation Scientifique du Departement dePathologie Animale[动物病理学部门组织的科学会议],INRA-ENV,Ecole Nationale Veterinaire de LYON(里昂国家畜医学校),13-14,1994年12月13-14日)借助于可在强启动子,即2型腺病毒主要晚期启动子的控制下表达gD基因的质粒,试着接种了猪以抗Aujeszkey病病毒。尽管抗体反应水平良好,但未显示保护作用。不过,用其中插入了gD基因和相同启动子的重组腺病毒接种猪之后,记录了有关保护作用的令人满意的结果,证明gD糖蛋白应足以在猪中诱导保护作用。
现有技术中通过多核苷酸接种方法在猪中未获得保护性结果。
因此,本发明打算提供多价的疫苗制剂,所述制剂可以确保接种猪能抵抗特别是涉及呼吸疾病和/或生殖疾病的多种致病因子。
本发明的另一目的是提供包含有不同效价并满足彼此相容性和效价稳定性所需的所有标准的疫苗制剂。
本发明的另一目的是提供可以在相同载体中含有不同效价的疫苗制剂。
本发明的另一目的是提供使用简单且便宜的疫苗制剂。
本发明的另一目的是提供这样一种疫苗制剂和接种猪的方法,所述疫苗和接种可以得到高效、长期以及安全性良好又不留后患的保护作用,包括多价保护作用。
因此,本发明的目的是特别地针对猪生殖和/或呼吸疾病的疫苗制剂,所述制剂含有至少3种效价的多核苷酸疫苗,每种含有一个整合质粒,其可在宿主细胞中体内表达一种猪病原体效价的基因,这些效价选自Aujeszkey病病毒(PRV或假狂犬病病毒),猪流感病毒(SIV),猪神秘病病毒(PRRS病毒),细小病毒病病毒(PPV病毒),典型猪瘟病毒(HCV病毒)和引起放线杆菌病的细菌(大叶性肺炎放线杆菌),对于每种效价而言,质粒含有一种或多种基因,该基因对于Aujeszkey病病毒选自gB和gD,对于猪流感病毒选自HA、NP和N,对于PRRS病毒选自ORF5(E),ORF3,ORF6(M),对于细小病毒病病毒选自VP2,对于典型猪瘟病毒选自E1、E2,对于大叶性肺炎放线杆菌选自apxI、apxII和apxIII。
本发明中的效价应理解成至少一种可提供抗所研究病原体病毒的保护作用的抗原,效价可含有得自一株或多株所研究病原体的一个或多个经修饰的天然基因作为亚效价。
病原性因子的基因应理解成不仅指完整的基因,也指不同的多种核苷酸序列,包括保持了诱导保护性反应之能力的片段。基因的概念覆盖了等价于实施例中详细描述的序列的核苷酸序列,也就是说,有所不同但编码相同蛋白质的序列。此概念也覆盖了被研究的其它株病原体的核苷酸序列,它可以提供交叉保护作用或特异于株或株群的保护作用。此概念也覆盖了已被修饰以便于由宿主动物在体内表达,但编码相同蛋白质的核苷酸序列。
优选本发明的疫苗制剂含有Aujeszkey和猪流感效价,在其中可加入优选选自PRRS和大叶性肺炎放线杆菌(放线杆菌病)效价的其它效价,任选在其中进入选自细小病毒病和典型猪瘟效价的其它效价。
当然,效价的所有联合都是可以的,然而,在本发明的范围内,Aujeszkey和猪流感,接下来是PRRS和大叶性肺炎放线杆菌的效价被认为是优选的。
从更特异地针对猪呼吸疾病的接种角度看,优选效价选自Aujeszkey、猪流感、PRRS和放线杆菌病。
从特异地针对猪生殖疾病的接种角度看,优选效价选自PRRS、细小病毒病、典型猪瘟和Aujeszkey病。
至于Aujeszkey效价,可使用gB或gD基因,优选使用这两种基因,此时所述基因位于不同质粒或一个相同质粒上。
至于猪流感效价,优选使用HA和NP基因,可以使用这两个基因中的一个,也可以同时使用这两个基因,此时所述基因位于不同质粒或一个相同质粒上。优选在同一疫苗中联合得自一种以上流感病毒株,尤其是得自在自然界发现的不同株的HA序列。另一方面,NP提供了交叉保护作用,因此得自单个病毒株的序列能令人满意。
至于PRSS效价,优选使用E和ORF3或M基因,可以单独或联合使用这些基因;联合使用时,基因可位于分离的质粒上,或位于联合有2或3个这种基因的质粒中。得自至少两株,尤其是得自欧洲株和美洲株的基因利于在同一疫苗中联合。
至于典型猪瘟效价,可使用E1和E2基因中的任一种或E1和E2基因,后情形下它们可在两个不同质粒或任选在一个相同质粒中联合。
至于放线杆菌病效价,可使用上述3种基因中的一个或位于不同质粒或混合质粒上的这些基因中2个或3个基因的联合以提供针对不同血清型大叶性肺炎放线杆菌的保护作用。对于apxI,II和III而言,可以设想修饰编码序列以得到解毒的抗原,具体方法见实施例。
特别对于经肌内途径接种而言,本发明疫苗制剂一般以0.1-10ml,特别是1-5ml的剂量体积形式提供。
剂量一般为每种质粒类型10ng-1mg,优选为100ng-50μg,优选为1μg-250μg。
优选使用简单地置于接种载体,一般为生理盐水(0.9%NaCl),超纯水,TE缓冲液等中的裸质粒。当然也可以使用现有技术中已描述的任何多核苷酸疫苗形式。
每个质粒都含有启动子,在其控制下可确保插入基因在宿主细胞中的表达。所述启动子一般是强的真核生物启动子,尤其是来源于人或鼠,或任选来源于其它动物如大鼠、猪和豚鼠的巨细胞病毒早期CMV-IE启动子。
启动子更一般地讲是来源于病毒或细胞。至于病毒启动子,可提到SV40病毒早期或晚期启动子或Rous肉瘤病毒LTR启动子。也可以是基因来源病毒的启动子,例如基因自身的启动子。
至于细胞启动子,可提到细胞骨架基因的启动子,例如结蛋白启动子(Bolmont等,亚显微镜细胞学和病理学(Journal of SubmicroscopicCytology and Pathology),1990,22,117-122;和Zhenlin等,基因(Gene),1989,78,243-254)或者肌动蛋白启动子。
当相同质粒中存在几个基因时,它们会存在于相同的转录单位或两个不同的单位中。
优选通过混合表达各种效价之抗原的多核苷酸质粒来达到本发明不同疫苗效价的联合,但也可以设想使几种效价的抗原由相同质粒表达。
本发明的目的还在于单价疫苗制剂,所述制剂含有一个或多个编码得自选自PRV,PRRS,PPV,HCV的病毒之一和大叶性肺炎放线杆菌的一个或多个基因的质粒,所述基因为上述的那些。除了它们的单价特征外,这些制剂可具有上述有关基因选择,基因联合,质粒组成,剂量体积,剂量等方面的特征。
单价疫苗制剂也可用于(i)制备上述的多价疫苗制剂,(ii)各对抗自己的疾病,(iii)与针对另一种疾病的另一类型(活的或灭活的完整的,重组的,亚单位)的疫苗联合,或(iv)按下述作为疫苗的加强剂。
实际上本发明另一个目的是使用一种或多种本发明的质粒以制备用于接种猪的疫苗,该猪首先已用第一种已知类型的常规疫苗接种,所述第一种疫苗特别选自活的完整疫苗,灭活的完整疫苗,亚单位疫苗和重组疫苗,所述第一种疫苗(单价或多价)具有(也就是说含有或能表达)由所述质粒编码的抗原或提供交叉保护作用的抗原。值得注意的是,该多核苷酸疫苗具有强有力的加强效果,可导致免疫应答的增强并可获得长期的免疫力。
一般讲,第一接种疫苗可选自可从不同的兽用疫苗生产商处购得的疫苗。
本发明的另一目的是接种试剂盒,所述试剂盒将上述第一接种疫苗和用于加强免疫的本发明的疫苗制剂组配在一起。本发明还涉及附带有说明书的本发明疫苗制剂,所述说明书指明该制剂可作为上述第一次接种的加强剂来使用。
本发明的另一目的是接种猪以抵抗猪生殖疾病和/或呼吸疾病的方法,所述方法包括施用有效量的上述疫苗制剂。此接种方法包括施用一剂或更多剂疫苗制剂,这些剂量可以在短时间内连续施用和/或以较长的间隔期连续施用。
在这种接种方法中,通过现有技术中建议的用于多核苷酸接种的不同施用途径并利用已知的施用技术施用本发明的疫苗制剂。特别可借助于液体注射器经液体喷射优选多个液体喷射经由皮内途径进行接种,尤其是使用具有几个孔或喷嘴,特别是具有5或6个孔或喷嘴的注射头的注射器,如法国里昂Endoscoptic公司制造并销售的Pigjet装置。
对于这种装置而言,优选剂量体积降低为0.1-0.9ml,特别是0.2-0.6ml,更优选为0.4-0.5ml,可以1次或几次,优选为2次施用该剂量体积。
本发明的另一目的是一种接种方法,所述方法包括按上述进行第一次接种,并用本发明的疫苗制剂加强接种。在本发明方法的优选实施方案中,首先给动物施用有效剂量的常规疫苗,尤其是灭活的、活的、减毒的或重组的、或亚单位的疫苗以提供第一次接种,一段时间后,优选为2至6周后,施用本发明的多价或单价疫苗。
本发明也涉及一种制备疫苗制剂的方法,即制备不同效价及其混合物的方法,如从此说明书中看到的方法。
下面参照附图,借助于本发明的实施方案更详细地描述本发明。
附图说明
图1:质粒pVR1012
图2:PRV gB基因的序列(NIA3株)
图3:质粒pAB090的构建
图4:PRV gD基因的序列(NIA3株)
图5:质粒pPB098的构建
图6:猪流感HA基因的序列(H1N1株)
图7:质粒pPB143的构建
图8:猪流感NP基因的序列(H1N1株)
图9:质粒pPB42的构建
图10:猪流感HA基因的序列(H3N2株)
图11:质粒pPB144的构建
图12:猪流感NP基因的序列(H3N2株)
图13:质粒pPB132的构建
图14:质粒pAB025
图15:质粒pAB001
图16:质粒pAB091
图17:质粒pAB092
图18:质粒pAB004
图19:质粒pAB069
图20:质粒pAB061
图21:质粒pPB162
图22:质粒pPB163
图23:质粒pPB174’
图24:质粒pPB189
图25:质粒pPB190
序列表SEQ ID NO:
SEQ ID NO:1:PRV gB基因的序列(NIA3株)
SEQ ID NO:2:寡核苷酸AB166
SEQ ID NO:3:寡核苷酸AB167
SEQ ID NO:4:寡核苷酸AB168
SEQ ID NO:5:寡核苷酸AB169
SEQ ID NO:6:PRV gD基因的序列(NIA3株)
SEQ ID NO:7:寡核苷酸PB101
SEQ ID NO:8:寡核苷酸PB102
SEQ ID NO:9:寡核苷酸PB107
SEQ ID NO:10:寡核苷酸PB108
SEQ ID NO:11:猪流感HA基因的序列(H1N1株)
SEQ ID NO:12:寡核苷酸PB097
SEQ ID NO:13:寡核苷酸PB098
SEQ ID NO:14:猪流感NP基因的序列(H1N1株)
SEQ ID NO:15:寡核苷酸PB095
SEQ ID NO:16:寡核苷酸PB096
SEQ ID NO:17:猪流感HA基因的序列(H3N2株)
SEQ ID NO:18:猪流感NP基因的序列(H3N2株)
SEQ ID NO:19:寡核苷酸AB055
SEQ ID NO:20:寡核苷酸AB056
SEQ ID NO:21:寡核苷酸AB001
SEQ ID NO:22:寡核苷酸AB002
SEQ ID NO:23:寡核苷酸AB170
SEQ ID NO:24:寡核苷酸AB171
SEQ ID NO:25:寡核苷酸AB172
SEQ ID NO:26:寡核苷酸AB173
SEQ ID NO:27:寡核苷酸AB007
SEQ ID NO:28:寡核苷酸AB010
SEQ ID NO:29:寡核苷酸AB126
SEQ ID NO:30:寡核苷酸AB127
SEQ ID NO:31:寡核苷酸AB118
SEQ ID NO:32:寡核苷酸AB119
SEQ ID NO:33:寡核苷酸PB174
SEQ ID NO:34:寡核苷酸PB189
SEQ ID NO:35:寡核苷酸PB190
SEQ ID NO:36:寡核苷酸PB175
SEQ ID NO:37:寡核苷酸PB176
SEQ ID NO:38:寡核苷酸PB191
SEQ ID NO:39:寡核苷酸PB192
SEQ ID NO:40:寡核苷酸PB177
SEQ ID NO:41:寡核苷酸PB278
SEQ ID NO:42:寡核苷酸PB279
SEQ ID NO:43:寡核苷酸PB280
SEQ ID NO:44:寡核苷酸PB307
SEQ ID NO:45:寡核苷酸PB303
SEQ ID NO:46:寡核苷酸PB306
SEQ ID NO:47:寡核苷酸PB304
SEQ ID NO:48:寡核苷酸PB305
实施例
实施例1:病毒的培养
在适当的细胞系统中培养病毒直至得到细胞病变效应。本领域技术人员熟知每种病毒所用的细胞系统。简单地说,用感染复数为1的被研究的病毒株接种对所用病毒敏感的细胞,所述细胞在Eagle最低基本培养基(MEM培养基)或另一种适当的培养基中培养。然后于37℃保温被感染的细胞直至出现完全的细胞病变效应(平均为36小时)。
实施例2:细菌的培养和细菌DNA的提取
按A.Rycroft等人(普通微生物学杂志(J.Gen.Microbiol.),1991,137,561-568)所述培养大叶性肺炎放线杆菌菌株。根据J.Sambrook等人(分子克隆:实验室手册,第2版,冷泉港实验室,冷泉港,纽约,1989)所述的标准技术制备高分子量的DNA(染色体DNA)。
实施例3:提取病毒基因组DNA
培养后,收获上清液和被裂解的细胞,于4℃,1000g将全部的病毒悬浮液离心10分钟以除去细胞碎片,然后通过于4℃,400,000g超速离心1小时以收获病毒颗粒。将沉淀物溶解于最少量的缓冲液(10mMTris,1mM EDTA;pH8.0)中,于37℃,在十二烷基硫酸钠(SDS)的存在下(终浓度为0.5%),用蛋白酶K(终浓度为100μg/ml)将上述浓缩的病毒悬浮液处理2小时。然后用苯酚/氯仿混合物提取病毒DNA,再用2倍体积的无水乙醇沉淀之。于-20℃放置过夜后,于4℃,10,000g将DNA离心15分钟,干燥DNA沉淀物,然后将它溶于最少量的无菌超纯水中,接着可以用限制性酶消化所述DNA。
实施例4:分离病毒基因组RNA
根据本领域技术人员熟知的技术纯化RNA病毒,然后使用P.Chromczynski和N.Sacchi所述的“硫氰酸胍/苯酚-氯仿”提取技术(分析生物化学,1987,162,156-159)分离各个病毒的基因组病毒RNA。
实施例5:分子生物学技术
使用J.Sambrook等人(分子克隆:实验室手册,第2版,冷泉港实验室,冷泉港,纽约,1989)所述的标准分子生物学技术进行所有的质粒构建。使用“Geneclean”试剂盒(BIO 101 Inc.La Jolla,CA)分离用于本发明的所有限制性片段。
实施例6:RT-PCR技术
合成多个特异性的寡核苷酸(在其5’末端含有限制性位点以便于克隆经扩增的片段)以使其完全覆盖欲被扩增的基因的编码区(见具体的实施例)。根据标准技术(J.Sambrook等人,分子克隆:实验室手册,第2版,冷泉港实验室,冷泉港,纽约,1989)进行逆转录(RT)反应和聚合酶链反应(PCR)。用一对特异性的扩增引物进行一种RT-PCR反应,所述反应的模板是提取出的病毒基因组RNA。用苯酚/氯仿/异戊醇(25∶24∶1)提取经扩增的互补DNA,然后用限制性酶消化。
实施例7:质粒pVR1012
质粒pVR1012(图1)得自Vical Inc.,San Diego,CA,USA。其构建描述于J.Hartikka等人(人类基因疗法(Human Gene Therapy),1996,7,1205-1217)。
实施例8:构建质粒pAB090(PRV gB基因)
用ApaI和NaeI消化质粒pPR2.15(M.Riviere等,病毒学杂志(J.Virol),1992,66,3424-3434),以释放出含有编码Aujeszky病病毒(NIA3株)gB糖蛋白之基因(图2和SEQ ID NO:1)的2665 bp的ApaI-NaeI片段(片段A)。
通过下列2个寡核苷酸的杂交:
AB166(33聚体)(SEQ ID NO:2)
5’GATGCCCGCTGGTGGCGGTCTTTGGCGCGGGCC 3’
AB167(33聚体)(SEQ ID NO:3)
5’ACGTCTACGGGCGACCACCGCCAGAAACCGCGC 3’
重新构建含有gD基因序列,始于起始的ATG密码子直至ApaI位点的33bp的片段(片段B),该片段在5’端产生了PstI位点。
通过下列2个寡核苷酸的杂交:
AB168(45聚体)(SEQ ID NO:4)
5’
GGCACTACCAGCGCCTCGAGAGCGAGGACCCCGACGCCCTG
TAGG 3’
AB169(49聚体)(SEQ ID NO:5)
5’
GATCCCTACAGGGCGTCGGGGTCCTCGCTCTCGAGGCGCTG
GTAGTGCC 3’
重新构建含有gD基因序列,始于NaeI位点直至TAG终止密码子的45bp的片段(片段C),该片段在3’端产生了BamHI位点。
将片断A,B和C一起连接到预先经PstI和BamHI消化的载体pVR1012(实施例7)中,得到质粒pAB090(7603 bp)(图3)。
实施例9:构建质粒pPB098(PRV gD基因)
用SalI和BglII消化质粒pPR29(M.Riviere等,病毒学杂志,1992,66,3424-3434),以释放出含有编码Aujeszky病病毒(NIA3株)gD糖蛋白之基因的3’部分(图4和SEQ ID NO:6)的711bp的SalI-BglII片段(片段A)。
用Eco47III和SalI消化质粒pPR29以释放出含有编码Aujeszky病病毒(NIA3株)gD糖蛋白之基因的5’部分的498 bp的Eco47III-SalI片段(片段B)。
通过下列2个寡核苷酸的杂交:
PB101(15聚体)(SEQ ID NO:7)
5’GATGCTGCTCGCAGC3’
PB102(19聚体)(SEQ ID NO:8)
5’GCTGCGAGCAGCATCTGCA 3’
重新构建含有gD基因5’序列,始于起始的ATG密码子直至Eco47III位点的15 bp的片段(片段C),该片段在5’端产生了PstI位点。
纯化之后,将片段A,B和C一起连接到预先经PstI和BglII消化的载体pVR1012(实施例7)中,得到质粒pPB098(6076 bp)(图5)。
实施例10:构建质粒pPB143(猪流感HA基因,H1N1株)
用根据实施例4所述技术制备的猪流感病毒(SIV H1N1“SW”株)基因组RNA,和下列寡核苷酸:
PB107(32聚体)(SEQ ID NO:9)
5’GTTCTGCAGCACCCGGGAGCAAAAGCAGGGGA 3’
PB108(33聚体)(SEQ ID NO:10)
5’ATTGCGGCCGCTAGTAGAAACAAGGGTGTTTTT 3’
根据实施例6所述技术进行RT-PCR反应以从SIV H1N1中精确地分离1803 bp PCR片段形式的编码HA蛋白的基因(图6和SEQ IDNO:11)。纯化之后,将此片段与载体PCRII-direct(Invitrogen ReferenceK2000-01)连接,得到载体pPB137(5755 bp)。用EcoRV和NotI消化载体pPB137以释放1820 bp的含有HA基因的EcoRV-NotI片段。将此片段与预先经EcoRV和NotI消化的载体pVR1012(实施例7)连接,得到质粒pPB143(6726 bp)(图7)。
实施例11:构建质粒pPB142(猪流感NP基因,H1N1株)
用根据实施例4所述技术制备的猪流感病毒(SIV H1N1“SW”株)基因组RNA,和下列寡核苷酸:
PB097(36聚体)(SEQ ID NO:12)
5’CCGGTCGACCGGGATAATCACTCACTGAGTGACATC 3’
PB098(33聚体)(SEQ ID NO:13)
5’TTGCGGCCGCTGTAGAAACAAGGGTATTTTTCT 3’
根据实施例6所述技术进行RT-PCR反应以从SIV H1N1中精确地分离SalI-NotI片段形式的编码NP蛋白的基因(图8和SEQ ID NO:14)。纯化之后,将1566 bp的RT-PCR产物与载体PCRII-direct(InvitrogenReference K2000-01)连接,得到载体pPB127(5519 bp)。
用SalI和NotI消化载体pPB127以释放1560 bp的含有NP基因的SalI-NotI片段。将此片段与预先经SalI和NotI消化的载体pVR1012(实施例7)连接,得到质粒pPB142(6451 bp)(图9)。
实施例12:构建质粒pPB144(猪流感HA基因,H3N2株)
用根据实施例4所述技术制备的猪流感病毒(SIV H3N2株,Cotesdu Nord 1987)基因组RNA,和下列寡核苷酸:
PB095(31聚体)(SEQ ID NO:15)
5’GTTCTGCAGGCAGGGGATAATTCTATCAACC 3’
PB096(36聚体)(SEQ ID NO:16)
5’TTGCGGCCGCAAGGGTGTTTTTAATTACTAATATAC 3’
根据实施例6所述技术进行RT-PCR反应以从SIV H3N2中精确地分离PstI-NotI片断形式的编码HA蛋白的基因(图10和SEQ IDNO:17)。纯化之后,将1765 bp的RT-PCR产物与载体PCRII-direct(Invitrogen Reference K2000-01)连接,得到载体pPB120(5716bp)。
用NotI消化载体pPB120以释放1797 bp的含有HA基因的NotI-NotI片段。将此片段与预先经NotI消化的载体pVR1012(实施例7)连接,得到质粒pPB144(6712 bp),该质粒在相对于启动子正确的方向上含有H3N2 HA基因(图11)。
实施例13:构建质粒pPB132(猪流感NP基因,H3N2株)
用根据实施例4所述技术制备的猪流感病毒(SIV H3N2株,Cotesdu Nord 1987)基因组RNA,和下列寡核苷酸:
PB097(36聚体)(SEQ ID NO:12)
5’CCGGTCGACCGGGATAATCACTCACTGAGTGACATC 3’
PB098(33聚体)(SEQ ID NO:13)
5’TTGCGGCCGCTGTAGAAACAAGGGTATTTTTCT 3’
根据实施例6所述技术进行RT-PCR反应以从SIV H3N2中精确地分离SalI-NotI片段形式的编码NP蛋白的基因(图12和SEQ ID NO:18)。纯化之后,将1564 bp的RT-PCR产物与载体PCRII-direct(InvitrogenReference K2000-01)连接,得到载体pPB123(5485 bp)。
用SalI和NotI消化载体pPB123以释放1558 bp的含有NP基因的SalI-NotI片段。将此片段与预先经SalI和NotI消化的载体pVR1012(实施例7)连接,得到质粒pPB132(6449 bp)(图13)。
实施例14:构建质粒pAB025(PRRSV ORF5基因,Lelystad株)
用根据实施例4所述技术制备的PRRSV病毒(Lelystad株)基因组RNA(J.Meulenberg等,病毒学(Virology),1993,19,62-72),和下列寡核苷酸:
AB055(34聚体)(SEQ ID NO:19)
5’ACGCGTCGACAATATGAGATGTTCTCACAAATTG 3’
AB056(33聚体)(SEQ ID NO:20)
5’CGCGGATCCCGTCTAGGCCTCCCATTGCTCAGC 3’
根据实施例6所述技术进行RT-PCR反应以从PRRS病毒Lelystad株中精确地分离编码包膜糖蛋白E(gp25)的“ORF5”基因。纯化之后,用SalI和BamHI消化630 bp的RT-PCR产物以分离617 bp的SalI-BamHI片段。将此片段与预先经SalI和BamHI消化的载体pVR1012(实施例7)连接,得到质粒pAB025(5486 bp)(图14)。
实施例15:构建质粒pAB001(PRRSV ORF5基因,USA株)
用根据实施例4所述技术制备的PRRSV病毒(ATCC VR2332株)基因组RNA(M.MuFtaugh等,Arch Virol,1995,140,1451-1460),和下列寡核苷酸:
AB001(30聚体)(SEQ ID NO:21)
5’AACTGCAGATGTTGGAGAAATGCTTGACCG 3’
AB002(30聚体)(SEQ ID NO:22)
5’CGGGATCCCTAAGGACGACCCCATTGTTCC 3’
根据实施例6所述技术进行RT-PCR反应以从PRRS病毒ATCC-VR2332株中精确地分离编码包膜糖蛋白E(gp25)的基因。纯化之后,用PstI和BamHI消化620 bp的RT-PCR产物以分离606 bp的PstI-BamHI片断。将此片段与预先经PstI和BamHI消化的载体pVR1012(实施例7)连接,得到质粒pAB001(5463 bp)(图15)。
实施例16:构建质粒pAB091(PRRSV ORF3基因,Lelystad株)
用根据实施例4所述技术制备的PRRSV病毒(Lelystad株)基因组RNA(J.Meulenberg等,病毒学,1993,19,62-72),和下列寡核苷酸:
AB170(32聚体)(SEQ ID NO:23)
5’AAACTGCAGCAATGGCTCATCAGTGTGCACGC 3’
AB171(30聚体)(SEQ ID NO:24)
5’CGCGGATCCTTATCGTGATGTACTGGGGAG 3’
根据实施例6所述技术进行RT-PCR反应以从PRRS病毒Lelystad株中精确地分离编码包膜糖蛋白“gp45”的“ORF3”基因。纯化之后,用PstI和BamHI消化818bp的RT-PCR产物以分离802bp的PstI-BamHI片断。将此片段与预先经PstI和BamHI消化的载体pVR1012(实施例7)连接,得到质粒pAB091(5660 bp)(图16)。
实施例17:构建质粒pAB092(PRRSV ORF3基因,USA株)
用根据实施例4所述技术制备的PRRSV病毒(ATCC VR2332株)基因组RNA(M.Murtaugh等,Arch Virol,1995,140,1451-1460),和下列寡核苷酸:
AN172(32聚体)(SEQ ID NO:25)
5’AAACTGCAGCAATGGTTAATAGCTGTACATTC 3’
AB173(32聚体)(SEQ ID NO:26)
5’CGCGGATCCCTATCGCCGTACGGCACTGAGGG 3’
根据实施例6所述技术进行RT-PCR反应以从PRRS病毒ATCC-VR2332株中精确地分离编码包膜糖蛋白“gp45”的“ORF3”基因。纯化之后,用PstI和BamHI消化785 bp的RT-PCR产物以分离769 bp的PstI-BamHI片段。将此片段与预先经PstI和BamHI消化的载体pVR1012(实施例7)连接,得到质粒pAB092(5627 bp)(图17)。
实施例18:构建质粒pAB004(猪细小病毒VP2基因)
用根据实施例4所述技术制备的猪细小病毒(NADL2株)基因组RNA(J.Vasudevacharya等,病毒学,1990,178,611-616),和下列寡核苷酸:
AB007(33聚体)(SEQ ID NO:27)
5’AAAACTGCAGAATGAGTGAAAATGTGGAACAAC 3’
AB010(33聚体)(SEQ ID NO:28)
5’CGCGGATCCCTAGTATAATTTTCTTGGTATAAG 3’
根据实施例6所述技术进行RT-PCR反应以扩增含有编码猪细小病毒VP2蛋白之基因的1757 bp的片段。纯化之后,用PstI和BamHI消化RT-PCR产物,得到1740 bp的PstI-BamHI片段。将此片段与预先经PstI和BamHI消化的载体pVR1012(实施例7)连接,得到质粒pAB004(6601bp)(图18)。
实施例19:构建质粒pAB069(猪瘟HCV E1基因)
用根据实施例4所述技术制备的猪瘟病毒(HCV)(Alfort株)基因组RNA(G.Meyers等,病毒学,1989,171,18-27),和下列寡核苷酸:
AB126(36聚体)(SEQ ID NO:29)
5’ACGCGTCGACATGAAACTAGAAAAAGCCCTGTTGGC 3’
AB127(34聚体)(SEQ ID NO:30)
5’CGCGGATCCTCATAGCCGCCCTTGTGCCCCGGTC 3’
根据实施例6所述技术进行RT-PCR反应以从HCV病毒中分离1363bp的RT-PCR片段形式的编码E1蛋白的序列。纯化之后,用SalI和BamHI消化此片段,得到1349 bp的SalI-BamHI片段。将此片段与预先经SalI和BamHI消化的载体pVR1012(实施例7)连接,得到质粒pAB069(6218 bp)(图19)。
实施例20:构建质粒pAB061(猪瘟HCV E2基因)
用根据实施例4所述技术制备的猪瘟病毒(HCV)(Alfort株)基因组RNA(G.Meyers等,病毒学,1989,171,18-27),和下列寡核苷酸:
AB118(36聚体)(SEQ ID NO:31)
5’ACGCGTCGACATGTCAACTACTGCGTTTCTCATTTG 3’
AB119(33聚体)(SEQ ID NO:32)
5’CGCGGATCCTCACTGTAGACCAGCAGCGAGCTG 3’
根据实施例6所述技术进行RT-PCR反应以从HCV病毒中分离1246bp的RT-PCR片段形式的编码E2蛋白的序列。纯化之后,用SalI和BamHI消化此片段,得到1232 bp的SalI-BamHI片段。将此片段与预先经SalI和BamHI消化的载体pVR1012(实施例7)连接,得到质粒pAB061(6101bp)(图20)。
实施例21:构建质粒pPB162(缺失的大叶性肺炎放线杆菌apxI基因)
克隆大叶性肺炎放线杆菌apxI基因以缺失氨基酸719-846之间富含甘氨酸的氨基酸区域(涉及钙离子的结合)。
用根据实施例2和3所述技术制备的大叶性肺炎放线杆菌(血清型1)的基因组DNA(J.Frey等,感染和免疫(Infect.Immun.),1991,59,3026-3032),和下列寡核苷酸:
PB174(32聚体)(SEQ ID NO:33)
5’TTGTCGACGTAAATAGCTAAGGAGACAACATG 3’
PB189(29聚体)(SEQ ID NO:34)
5’TTGAATTCTTCTTCAACAGAATGTAATTC3’
进行PCR反应以扩增SalI-EcoRI片段形式的编码大叶性肺炎放线杆菌溶血素I蛋白之apxI基因的5’部分。纯化之后,用SalI和EcoRI消化2193 bp的PCR产物以分离2183 bp的SalI-EcoRI片段(片段A)。
用大叶性肺炎放线杆菌(血清型1)的基因组DNA(J.Frey等,
Infect.Immun.,1991,59,3026-3032),和下列寡核苷酸:
PB190(32聚体)(SEQ ID NO:35)
5’TTGAATTCTATCGCTACAGTAAGGAGTACGG 3’
PB175(31聚体)(SEQ ID NO:36)
5’TTGGATCCGCTATTTATCATCTAAAAATAAC 3’
进行PCR反应以扩增EcoRI-BamHI片段形式的编码大叶性肺炎放线杆菌溶血素I蛋白之apxI基因的3’部分。纯化之后,用EcoRI和BamHI消化576 bp的PCR产物以分离566 bp的EcoRI-BamHI片段(片段B)。将片段A和B与预先经SalI和BamHI消化的载体pVR1012(实施例7)连接,得到质粒pPB162(7619 bp)(图21)。
实施例22:构建质粒pPB163(缺失的大叶性肺炎放线杆菌apxII基因)
克隆大叶性肺炎放线杆菌apxII基因以缺失氨基酸716-813之间富含甘氨酸的氨基酸区域(涉及钙离子的结合)。
用根据实施例2和3所述技术制备的大叶性肺炎放线杆菌(血清型9)的基因组DNA(M.Smits等,感染和免疫(Infection andImmunity),1991,59,4497-4504),和下列寡核苷酸:
PB176(31聚体)(SEQ ID NO:37)
5’TTGTCGACGATCAATTATATAAAGGAGACTC 3’
PB191(30聚体)(SEQ ID NO:38)
5’TTGAATTCCTCTTCAACTGATTTGAGTGAG 3’
进行PCR反应以扩增SalI-EcoRI片段形式的编码大叶性肺炎放线杆菌溶血素II蛋白之apxII基因的5’部分。纯化之后,用SalI和EcoRI消化2190 bp的PCR产物以分离2180 bp的SalI-EcoRI片段(片段A)。
用大叶性肺炎放线杆菌(血清型9)的基因组DNA(M.Smits等,感染和免疫,1991,59,4497-4504),和下列寡核苷酸:
PB192(29聚体)(SEQ ID NO:39)
5’TTGAATTCGTAAATCTTAAAGACCTCACC 3’
PB177(30聚体)(SEQ ID NO:40)
5’TTGGATCCACCATAGGATTGCTATGATTTG 3’
进行PCR反应以扩增EcoRI-BamHI片段形式的编码大叶性肺炎放线杆菌溶血素II蛋白之apxII基因的3’部分。纯化之后,用EcoRI和BamHI消化473 bp的PCR产物以分离463 bp的EcoRI-BamHI片段(片段B)。
将片段A和B与预先经SalI和BamHI消化的载体pVR1012(实施例7)连接,得到质粒pPB163(7513 bp)(图22)。
实施例23:构建质粒pPB174’,pPB189和pPB190(缺失的大叶性肺炎放线杆菌apxIII基因)
apxIII缺失的第一个实施例(质粒pPB174’)
克隆大叶性肺炎放线杆菌apxIII基因以缺失氨基酸733-860之间富含甘氨酸的氨基酸区域(涉及钙离子的结合)。
用根据实施例2和3所述技术制备的大叶性肺炎放线杆菌(血清型8)的基因组DNA(M.Smits,1992,Genbank序列登记号为X68815),和下列寡核苷酸:
PB278(30聚体)(SEQ ID NO:41)
5’TTTGTCGACATGAGTACTTGGTCAAGCATG 3’
PB279(28聚体)(SEQ ID NO:42)
5’TTTATCGATTCTTCTACTGAATGTAATTC 3’
进行PCR反应以扩增SalI-ClaI片段形式的apxIII基因(编码大叶性肺炎放线杆菌溶血素III蛋白)的5’部分。纯化之后,用SalI和ClaI消化2216 bp的PCR产物以分离2205 bp的SalI-ClaI片段(片段A)。
用大叶性肺炎放线杆菌(血清型8)的基因组DNA(M.Smits,1992,Genbank序列登记号为X68815),和下列寡核苷酸:
PB280(33聚体)(SEQ ID NO:43)
5’TTTATCGATTTATGTTTATCGTTCCACTTCAGG 3’
PB307(32聚体)(SEQ ID NO:44)
5’TTGGATCCTTAAGCTGCTCTAGCTAGGTTACC 3’
进行PCR反应以扩增ClaI-BamHI片段形式的apxIII基因(编码大叶性肺炎放线杆菌溶血素III蛋白)的3’部分。纯化之后,用ClaI和BamHI消化596 bp的PCR产物以分离583 bp的ClaI-BamHI片段(片段B)。将片段A和B与预先经SalI和BamHI消化的载体pVR1012(实施例7)连接,得到质粒pPB174’(7658 bp)(图23)。
apxIII缺失的第二个实施例(质粒pPB189)
克隆大叶性肺炎放线杆菌apxIII基因以缺失氨基酸705-886之间富含甘氨酸的氨基酸区域(涉及钙离子的结合)。
用根据实施例2和3所述技术制备的大叶性肺炎放线杆菌(血清型8)的基因组DNA(M.Smits,1992,Genbank序列登记号为X68815),和下列寡核苷酸:
PB278(30聚体)(SEQ ID NO:41)
5’TTTGTCGACATGAGTACTTGGTCAAGCATG 3’
PB303(32聚体)(SEQ ID NO:45)
5’TTTATCGATTTCTTCACGTTTACCAACAGCAG 3’
进行PCR反应以扩增SalI-ClaI片段形式的apxIII基因(编码大叶性肺炎放线杆菌溶血素III蛋白)的5’部分。纯化之后,用SalI和ClaI消化2133bp的PCR产物以分离2122 bp的SalI-ClaI片段(片段A)。
用大叶性肺炎放线杆菌(血清型8)的基因组DNA(M.Smits,1992,Genbank序列登记号为X68815),和下列寡核苷酸:
PB306(31聚体)(SEQ ID NO:46)
5’TTTATCGATTCTGATTTTTCCTTCGATCGTC 3’
PB307(32聚体)(SEQ ID NO:44)
5’TTGGATCCTTAAGCTGCTCTAGCTAGGTTACC 3’
进行PCR反应以扩增ClaI-BamHI片段形式的apxIII基因(编码大叶性肺炎放线杆菌溶血素III蛋白)的3’部分。纯化之后,用ClaI和BamHI消化518 bp的PCR产物以分离506 bp的ClaI-BamHI片段(片段B)。将片段A和B与预先经SalI和BamHI消化的载体pVR1012(实施例7)连接,得到质粒pPB189(7496 bp)(图24)。
apxIII缺失的第三个实施例(质粒pPB190)
克隆大叶性肺炎放线杆菌apxIII基因以缺失氨基酸718-876之间富含甘氨酸的氨基酸区域(涉及钙离子的结合)。
用根据实施例2和3所述技术制备的大叶性肺炎放线杆菌(血清型8)的基因组DNA(M.Smits,1992,Genbank序列登记号为X68815),和下列寡核苷酸:
PB278(30聚体)(SEQ ID NO:41)
5’TTTGTCGACATGAGTACTTGGTCAAGCATG 3’
PB304(33聚体)(SEQ ID NO:47)
5’TTTATCGATACCTGATTGCGTTAATTCATAATC 3’
进行PCR反应以扩增SalI-ClaI片段形式的apxIII基因(编码大叶性肺炎放线杆菌溶血素III蛋白)的5’部分。纯化之后,用SalI和ClaI消化2172 bp的PCR产物以分离2161 bp的SalI-ClaI片段(片段A)。
用大叶性肺炎放线杆菌(血清型8)的基因组DNA(M.Smits,1992,Genbank序列登记号为X68815),和下列寡核苷酸:
PB305(31聚体)(SEQ ID NO:48)
5’TTTATCGATAAATCTAGTGATTTAGATAAAC 3’
PB307(32聚体)(SEQ ID NO:44)
5’TTGGATCCTTAAGCTGCTCTAGCTAGGTTACC 3’
进行PCR反应以扩增ClaI-BamHI片段形式的apxIII基因(编码大叶性肺炎放线杆菌溶血素III蛋白)的3’部分。纯化之后,用ClaI和BamHI消化548 bp的PCR产物以分离536 bp的ClaI-BamHI片段(片段B)。将片段A和B与预先经SalI和BamHI消化的载体pVR1012(实施例7)连接,得到质粒pPB190(7565 bp)(图25)。
实施例24:制备和纯化质粒
为了制备欲接种动物的质粒,可使用能得到超螺旋形式占优势的纯化质粒悬浮液的任何技术,这些技术是本领域技术人员所熟知的。这里可特别提到J.Sambrook等人(分子克隆:实验室手册,第2版,冷泉港实验室,冷泉港,纽约,1989)所述的碱裂解技术,接着再在溴化乙锭的存在下,在氯化铯梯度中进行2次连续的超速离心。也可参照专利申请PCT WO 95/21250和PCT WO 96/02658,它们描述了工业化规模生产可用于接种之质粒的方法。为了制备疫苗(见实施例17),可重新悬浮纯化的质粒以得到适于储存的高浓度溶液(>2mg/ml)。为此,将质粒重新悬浮于超纯水或TE缓冲液(10mM Tris-HCl;1mM EDTA,pH8.0)中。
实施例25:制备联合疫苗
从其浓缩溶液(实施例16)开始混合制备联合疫苗所需的多种质粒。制备混合物以使各个质粒的终浓度相当于各个质粒的有效剂量。可被用于调节疫苗终浓度的溶液可以是0.9%的NaCl溶液,也可以是PBS缓冲液。
也可以将特殊的制剂,如脂质体,阳离子脂质用于制备疫苗。
实施例26:对猪的接种
以每种质粒100,250或500μg的剂量接种猪。
通过肌内途径用针头进行注射。此时,以2ml的体积施用疫苗剂量。
也可使用在5个点分送0.2ml剂量(每个注射点0.04ml)的液体喷射注射器装置(不带针头)(如“PIGJET”装置),经由皮内途径进行注射。此时,施用的疫苗剂量体积为0.2或0.4ml,这分别相应于注射1次或2次。当利用PIGJET装置进行2次连续的注射时,应将注射区间隔开以使2个注射区彼此间隔约1-2cm的距离。
Claims (7)
1.一种猪疫苗,其中包含含有Aujeszky病病毒基因的质粒,所述基因选自gB和gD。
2.根据权利要求1的疫苗,其中所述质粒包含gB和gD基因二者。
3.根据权利要求1的疫苗,其中所述质粒包含gB基因。
4.根据权利要求1的疫苗,其中所述质粒包含gD基因。
5.根据权利要求1的疫苗,其中包含含有gB基因的质粒和含有gD基因的质粒。
6.根据权利要求1-5任一项的疫苗,其中所述质粒包含巨细胞病毒CMV-IE启动子。
7.根据权利要求1-5任一项的疫苗,其中所述质粒包含选自SV40病毒早期启动子、SV40病毒晚期启动子、劳氏肉瘤病毒LTR启动子和细胞骨架基因启动子的启动子。
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Families Citing this family (63)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
AU762901B2 (en) * | 1996-07-19 | 2003-07-10 | Merial | Polynucleotide vaccine formula for treating porcine respiratory and reproductive diseases |
EP0863151A1 (en) * | 1997-02-12 | 1998-09-09 | Akzo Nobel N.V. | "Canine parvovirus dna vaccines" |
CA2223029A1 (en) | 1997-02-12 | 1998-08-12 | Akzo Nobel Nv | Canine parvovirus dna vaccines |
US7211379B2 (en) * | 1997-10-03 | 2007-05-01 | Merial Sas | Prevention of myocarditis, abortion and intrauterine infection associated with porcine circovirus-2 |
US7192594B2 (en) * | 1997-10-03 | 2007-03-20 | Merial Limited | Postweaning multisystemic wasting syndrome and porcine circovirus from pigs |
FR2776294B1 (fr) * | 1998-03-20 | 2001-06-22 | Merial Sas | Nouveaux circovirus porcins ; vaccins et reactifs de diagnostic |
FR2781159B1 (fr) * | 1998-07-06 | 2000-10-06 | Merial Sas | Vaccin circovirus et parvovirus porcin |
US6517843B1 (en) | 1999-08-31 | 2003-02-11 | Merial | Reduction of porcine circovirus-2 viral load with inactivated PCV-2 |
FR2772047B1 (fr) * | 1997-12-05 | 2004-04-09 | Ct Nat D Etudes Veterinaires E | Sequence genomique et polypeptides de circovirus associe a la maladie de l'amaigrissement du porcelet (map), applications au diagnostic et a la prevention et/ou au traitement de l'infection |
DK2363488T3 (en) * | 1997-12-11 | 2014-11-24 | Merial Sas | Postweaning multisystemic wasting syndrome virus from pigs |
ES2170622B1 (es) * | 1999-12-03 | 2004-05-16 | Consejo Superior De Investigaciones Cientificas | Clones y vectores infectivos derivados de coronavirus y sus aplicaciones. |
FR2804028B1 (fr) * | 2000-01-21 | 2004-06-04 | Merial Sas | Vaccins adn ameliores pour animaux de rente |
KR20030054139A (ko) * | 2001-12-24 | 2003-07-02 | 학교법인 건국대학교 | 돼지 오제스키병 바이러스의 gD 유전자를 포함하는재조합 발현 벡터 및 이를 이용한 DNA 백신 |
US7906311B2 (en) | 2002-03-20 | 2011-03-15 | Merial Limited | Cotton rat lung cells for virus culture |
US20060160759A1 (en) * | 2002-09-28 | 2006-07-20 | Jianzhu Chen | Influenza therapeutic |
EP1606419A1 (en) | 2003-03-18 | 2005-12-21 | Quantum Genetics Ireland Limited | Systems and methods for improving protein and milk production of dairy herds |
US7468273B2 (en) | 2003-05-01 | 2008-12-23 | Meial Limited | Canine GHRH gene, polypeptides and methods of use |
WO2005049794A2 (en) | 2003-11-13 | 2005-06-02 | University Of Georgia Research Foundation, Inc. | Methods of characterizing infectious bursal disease virus |
US7527967B2 (en) * | 2003-11-25 | 2009-05-05 | Academia Sinica | Recombinant baculovirus and virus-like particle |
ATE510928T1 (de) | 2004-02-19 | 2011-06-15 | Univ Alberta | Leptinpromotor-polymorphismen und verwendungen davon |
ES2343270T3 (es) | 2005-04-25 | 2010-07-27 | Merial Ltd. | Vacunas contra el virus nipah. |
US20080241184A1 (en) | 2005-08-25 | 2008-10-02 | Jules Maarten Minke | Canine influenza vaccines |
EP3147296A1 (en) | 2005-11-14 | 2017-03-29 | Merial, Inc. | Gene therapy for renal failure |
US7771995B2 (en) | 2005-11-14 | 2010-08-10 | Merial Limited | Plasmid encoding human BMP-7 |
US7862821B2 (en) | 2006-06-01 | 2011-01-04 | Merial Limited | Recombinant vaccine against bluetongue virus |
KR101445903B1 (ko) | 2007-05-02 | 2014-09-29 | 메리얼 리미티드 | 향상된 발현 및 안정성을 갖는 dna 플라스미드 |
US20080274137A1 (en) * | 2007-05-02 | 2008-11-06 | Jean Christophe Francis Audonnet | DNA plasmids having improved expression and stability |
EP2367566B2 (en) | 2008-11-28 | 2019-11-27 | Boehringer Ingelheim Animal Health USA Inc. | Recombinant avian influenza vaccine and uses thereof |
CN102428099B (zh) | 2009-04-03 | 2016-03-16 | 梅里亚有限公司 | 运载新城疫病毒的禽疫苗 |
CA2785653C (en) | 2009-12-28 | 2018-05-01 | Merial Limited | Recombinant ndv antigen and uses thereof |
US20130197612A1 (en) | 2010-02-26 | 2013-08-01 | Jack W. Lasersohn | Electromagnetic Radiation Therapy |
CN103492410A (zh) | 2010-03-12 | 2014-01-01 | 梅里亚有限公司 | 蓝舌病毒重组疫苗和其使用 |
CN102370976B (zh) * | 2010-08-09 | 2014-06-11 | 中山大学 | 猪流感病毒和猪口蹄疫病毒的混合病毒样颗粒、制备方法和应用 |
CN102373181B (zh) * | 2010-08-09 | 2014-06-11 | 中山大学 | 猪流感病毒和猪蓝耳病病毒混合病毒样颗粒、制备方法和应用 |
MX344103B (es) | 2010-08-31 | 2016-12-05 | Merial Ltd | Vacunas de virus herpes vectorizado con virus de enfermedad de newcastle. |
WO2012090073A2 (en) | 2010-12-30 | 2012-07-05 | The Netherlands Cancer Institute | Methods and compositions for predicting chemotherapy sensitivity |
EP2694678A2 (en) | 2011-04-04 | 2014-02-12 | Netherland Cancer Institute | Methods and compositions for predicting resistance to anticancer treatment |
AU2012240240A1 (en) | 2011-04-04 | 2013-05-09 | Netherlands Cancer Institute | Methods and compositions for predicting resistance to anticancer treatment with protein kinase inhibitors |
US9216213B2 (en) | 2011-04-20 | 2015-12-22 | Merial, Inc. | Adjuvanted rabies vaccine with improved viscosity profile |
CN103945864A (zh) | 2011-04-25 | 2014-07-23 | 先进生物学实验室股份有限公司 | 截短的hiv包膜蛋白(env)、其相关方法和组合物 |
BR112013028605A2 (pt) | 2011-05-07 | 2017-01-17 | Avi Mex S A De C V Lab | vetor viral, vacina recombinante contra prrs e uso de uma vacina |
BR112013030321A2 (pt) * | 2011-05-27 | 2017-07-11 | Sinovet Beijing Biotechnology Co Ltd | composição de vacina, método para preparar a composição de vacina, uso da composição de vacina, método para imunizar um porco, cepa de vacina contra csfv e uso de uma célula no cultivo de uma cepa de vacina contra csfv. |
CA2837582C (en) | 2011-05-27 | 2021-03-02 | Merial Limited | Genetic vaccines against hendra virus and nipah virus |
US9669085B2 (en) | 2011-06-01 | 2017-06-06 | Merial Inc. | Needle-free administration of PRRSV vaccines |
CN103841963B (zh) | 2011-08-12 | 2018-04-24 | 梅里亚股份有限公司 | 用于生物产品特别是疫苗的真空辅助保存 |
WO2013093629A2 (en) | 2011-12-20 | 2013-06-27 | Netherlands Cancer Institute | Modular vaccines, methods and compositions related thereto |
NZ628270A (en) | 2012-02-14 | 2016-09-30 | Merial Inc | Recombinant poxviral vectors expressing both rabies and ox40 proteins, and vaccines made therefrom |
MX353786B (es) | 2012-02-14 | 2018-01-29 | Merial Inc | Vacunas de subunidades de rotavirus, metodos de preparacion y uso de las mismas. |
WO2013138776A1 (en) | 2012-03-16 | 2013-09-19 | Merial Limited | Novel methods for providing long-term protective immunity against rabies in animals, based upon administration of replication-deficient flavivirus expressing rabies g |
EP2861248B1 (en) | 2012-06-13 | 2017-11-15 | Merial, Inc. | Reassortant btv and ahsv vaccines |
CN102807989B (zh) * | 2012-08-01 | 2014-04-23 | 中国人民解放军军事医学科学院军事兽医研究所 | 犬科和/或猫科动物疫病重组活载体疫苗的制备方法 |
EP2968514A1 (en) | 2013-03-12 | 2016-01-20 | Merial, Inc. | Reverse genetics schmallenberg virus vaccine compositions, and methods of use thereof |
US20160137700A1 (en) * | 2013-05-31 | 2016-05-19 | Pulike Biological Engineering, Inc. | Porcine pseudorabies virus, vaccine composition and preparation method and use thereof |
WO2015048115A1 (en) | 2013-09-25 | 2015-04-02 | Zoetis Llc | Pcv2b divergent vaccine composition and methods of use |
CN105251000B (zh) * | 2014-09-30 | 2018-12-14 | 普莱柯生物工程股份有限公司 | 猪伪狂犬病病毒疫苗组合物及其制备方法和应用 |
NZ731659A (en) | 2014-11-03 | 2018-10-26 | Merial Inc | Methods of using microneedle vaccine formulations to elicit in animals protective immunity against rabies virus |
PL3313864T3 (pl) | 2015-06-23 | 2022-01-03 | Boehringer Ingelheim Animal Health USA Inc. | Rekombinowane wektory wirusowe zawierające drugorzędne białko prrsv oraz sposoby ich wytwarzania i zastosowania |
CN105693827B (zh) * | 2015-06-29 | 2020-05-15 | 普莱柯生物工程股份有限公司 | 一种猪伪狂犬病病毒亚单位疫苗及其制备方法和应用 |
CN104998256A (zh) * | 2015-07-14 | 2015-10-28 | 天津瑞普生物技术股份有限公司 | 一种猪用三联灭活疫苗的制备方法 |
CA2996143A1 (en) | 2015-08-20 | 2017-02-23 | Merial, Inc. | Fcv recombinant vaccines and uses thereof |
FI3355915T3 (fi) | 2015-09-29 | 2024-01-12 | Boehringer Ingelheim Animal Health Usa Inc | Koirien parvoviruksen (CPV) viruksen kaltaisten hiukkasten (VLP) rokotteet ja niiden käytöt |
KR102153303B1 (ko) | 2015-11-23 | 2020-09-09 | 뵈링거 잉겔하임 애니멀 헬스 유에스에이 인코포레이티드 | Fmdv 및 e2 융합 단백질 및 이의 용도 |
TWI760322B (zh) | 2016-01-29 | 2022-04-11 | 美商百靈佳殷格翰動物保健美國有限公司 | 重組腺病毒載體裝載之fmdv疫苗及其用途 |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1995020660A2 (en) * | 1994-01-27 | 1995-08-03 | University Of Massachusetts Medical Center | Immunization by inoculation of dna transcription unit |
WO1996006619A1 (en) * | 1994-09-01 | 1996-03-07 | Paul Prem S | Polynucleic acids and proteins from a porcine reproductive and respiratory syndrome virus and uses thereof |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
NL8901651A (nl) * | 1989-06-29 | 1991-01-16 | Centraal Diergeneeskundig Inst | Vaccin tegen pestivirusinfecties, zoals varkenspest; daarvoor bruikbare nucleotidesequenties en polypeptiden. |
US5516905A (en) * | 1994-08-30 | 1996-05-14 | University Of Massachusetts Medical Center | Antibiotic compounds and methods to treat gram-positive bacterial and mycoplasmal infections |
CA2216308A1 (en) * | 1995-12-21 | 1997-07-03 | Solvay (Societe Anonyme) | Plasmid vaccine for pseudorabies virus |
-
1996
- 1996-07-19 FR FR9609338A patent/FR2751224B1/fr not_active Expired - Lifetime
-
1997
- 1997-07-11 AR ARP970103116A patent/AR007864A1/es unknown
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2001
- 2001-02-16 US US09/784,984 patent/US6576243B1/en not_active Expired - Lifetime
-
2002
- 2002-12-09 RU RU2002133052/13A patent/RU2305559C2/ru active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1995020660A2 (en) * | 1994-01-27 | 1995-08-03 | University Of Massachusetts Medical Center | Immunization by inoculation of dna transcription unit |
WO1996006619A1 (en) * | 1994-09-01 | 1996-03-07 | Paul Prem S | Polynucleic acids and proteins from a porcine reproductive and respiratory syndrome virus and uses thereof |
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CA2261346A1 (en) | 1998-01-29 |
RU2002133052A (ru) | 2005-01-20 |
DE69734284T2 (de) | 2006-07-13 |
DE69734284D1 (de) | 2006-02-09 |
ES2251030T3 (es) | 2006-04-16 |
HUP9903822A2 (hu) | 2000-03-28 |
WO1998003658A1 (fr) | 1998-01-29 |
DK0912743T3 (da) | 2006-01-30 |
EP0912743B1 (fr) | 2005-09-28 |
UA77935C2 (uk) | 2007-02-15 |
FR2751224B1 (fr) | 1998-11-20 |
PL190615B1 (pl) | 2005-12-30 |
AU3699197A (en) | 1998-02-10 |
US6576243B1 (en) | 2003-06-10 |
KR20000065256A (ko) | 2000-11-06 |
TW589190B (en) | 2004-06-01 |
JP2001500111A (ja) | 2001-01-09 |
BR9710740A (pt) | 1999-08-17 |
FR2751224A1 (fr) | 1998-01-23 |
US6207165B1 (en) | 2001-03-27 |
EP0912743A1 (fr) | 1999-05-06 |
HUP9903822A3 (en) | 2010-01-28 |
PL331249A1 (en) | 1999-07-05 |
KR100496249B1 (ko) | 2006-01-27 |
UA91675C2 (ru) | 2010-08-25 |
CO4750676A1 (es) | 1999-03-31 |
RU2305559C2 (ru) | 2007-09-10 |
AU735291B2 (en) | 2001-07-05 |
CN1225684A (zh) | 1999-08-11 |
CA2261346C (en) | 2012-01-03 |
CN101121023A (zh) | 2008-02-13 |
AR007864A1 (es) | 1999-11-24 |
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