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31. 发明授权

US07829102B2 Production of attenuated, human-bovine chimeric respiratory syncytial virus vaccines 失效
标题翻译：减毒，人 - 牛嵌合呼吸道合胞病毒疫苗的生产
公开(公告)号：US07829102B2
公开(公告)日：2010-11-09
申请号：US11097946
申请日：2005-03-31
申请人： Ursula Buchholz , Peter L. Collins , Brian R. Murphy , Stephen S. Whitehead , Christine D. Krempl
发明人： Ursula Buchholz , Peter L. Collins , Brian R. Murphy , Stephen S. Whitehead , Christine D. Krempl
IPC分类号： A61K39/155
CPC分类号： C12N7/00 , A61K39/00 , A61K39/12 , A61K39/155 , A61K2039/5254 , A61K2039/5256 , A61K2039/544 , C07K14/005 , C07K2319/00 , C12N15/86 , C12N2760/18522 , C12N2760/18534 , C12N2760/18543 , C12N2760/18561 , C12N2840/203
摘要： Chimeric human-bovine respiratory syncytial virus (RSV) are infectious and attenuated in humans and other mammals and useful in vaccine formulations for eliciting an anti-RSV immune response. Also provided are isolated polynucleotide molecules and vectors incorporating a chimeric RSV genome or antigenome which includes a partial or complete human or bovine RSV “background” genome or antigenome combined or integrated with one or more heterologous gene(s) or genome segment(s) of a different RSV strain. Chimeric human-bovine RSV of the invention include a partial or complete “background” RSV genome or antigenome derived from or patterned after a human or bovine RSV strain or subgroup virus combined with one or more heterologous gene(s) or genome segment(s) of a different RSV strain or subgroup virus to form the human-bovine chimeric RSV genome or antigenome.
摘要翻译：嵌合人 - 牛呼吸道合胞病毒（RSV）在人类和其他哺乳动物中具有感染性和减毒性，并且可用于引发抗RSV免疫应答的疫苗制剂中。还提供了分离的多核苷酸分子和掺入嵌合RSV基因组或抗原组的载体，其包括部分或完整的人或牛RSV“背景”基因组或与一个或多个异源基因或基因组片段不同的RSV菌株。本发明的嵌合人类牛RSV包括部分或完整的“背景”RSV基因组或在与一个或多个异源基因或基因组片段组合的人或牛RSV病毒株或亚组病毒之后衍生或构图的部分或完整的“背景” 的不同RSV株或亚组病毒形成人 - 牛嵌合RSV基因组或抗原组。

32. 发明授权

US07485440B2 Production of attenuated respiratory syncytial virus vaccines involving modification of M2 ORF2 有权
标题翻译：涉及M2 ORF2修饰的减毒呼吸道合胞病毒疫苗的生产
公开(公告)号：US07485440B2
公开(公告)日：2009-02-03
申请号：US11011502
申请日：2004-12-13
申请人： Peter L. Collins , Brian R. Murphy , Alison Bermingham
发明人： Peter L. Collins , Brian R. Murphy , Alison Bermingham
IPC分类号： C12P21/06 , A61K39/155
CPC分类号： A61K39/155 , A61K39/12 , A61K2039/5254 , A61K2039/543 , A61K2039/544 , C07K14/005 , C12N7/00 , C12N2760/18522 , C12N2760/18534 , C12N2760/18561
摘要： Recombinant respiratory syncytial virus (RSV) are provided in which expression of the second translational open reading frame encoded by the M2 gene (M2ORF2) is reduced or ablated to yield novel RSV vaccine candidates. Expression of M2 ORF2 is reduced or ablated by modifying a recombinant RSV genome or antigenome to incorporate a frame shift mutation, or one or more stop codons in M2 ORF2. Alternatively, M2 ORF2 is deleted in whole or in part to render the M2-2 protein partially or entirely non-functional or to disrupt its expression altogether. M2 ORF2 deletion and knock out mutants possess highly desirable phenotypic characteristics for vaccine development. These changes specify one or more desired phenotypic changes in the resulting virus or subviral particle. Vaccine candidates are generated that show a change in mRNA transcription, genomic or antigenomic RNA replication, viral growth characteristics, viral antigen expression, viral plaque size, and/or a change in cytopathogenicity. In addition, M2-2 knock out or deletion virus exhibits increased levels of synthesis of viral proteins in cell culture, providing an enriched source of viral antigen or protein for purification and use as a noninfectious subunit vaccine.
摘要翻译：提供了重组呼吸道合胞病毒（RSV），其中由M2基因（M2ORF2）编码的第二翻译开放阅读框的表达被减少或消融以产生新的RSV疫苗候选物。 M2 ORF2的表达通过修饰重组RSV基因组或抗原组合来结合帧移位突变或M2 ORF2中的一个或多个终止密码子来减少或消除。或者，M2 ORF2全部或部分缺失，以使M2-2蛋白部分或完全不起作用或完全破坏其表达。 M2 ORF2缺失和敲除突变体对于疫苗开发具有非常需要的表型特征。这些变化指定所得病毒或亚病毒颗粒中的一种或多种期望的表型变化。产生疫苗候选物，其显示mRNA转录，基因组或反基因组RNA复制，病毒生长特征，病毒抗原表达，病毒斑块大小和/或细胞致病性变化的变化。此外，M2-2敲除或缺失病毒在细胞培养物中表现出增加的病毒蛋白质合成水平，提供用于纯化和用作非感染性亚单位疫苗的病毒抗原或蛋白质的丰富来源。

33. 发明授权

US07846455B2 Attenuated chimeric respiratory syncytial virus 有权
标题翻译：减毒嵌合呼吸道合胞病毒
公开(公告)号：US07846455B2
公开(公告)日：2010-12-07
申请号：US10722000
申请日：2003-11-25
申请人： Peter L. Collins , Brian R. Murphy , Stephen S. Whitehead
发明人： Peter L. Collins , Brian R. Murphy , Stephen S. Whitehead
IPC分类号： A61K39/155 , A61K39/12 , C12N7/00 , C12N7/01 , C12N7/04
CPC分类号： C07K14/005 , A61K39/00 , A61K2039/5254 , C12N7/00 , C12N2760/18522 , C12N2760/18543 , C12N2760/18561
摘要： Chimeric respiratory syncytial virus (RSV) and vaccine compositions thereof are produced by introducing one or more heterologous gene(s) or gene segment(s) from one RSV subgroup or strain into a recipient RSV backround of a different subgroup or strain. The resulting chimeric RSV virus or subviral particle is infectious and attenuated, preferably by introduction of selected mutations specifying attenuated phenotypes into a chimeric genome or antigenome to yield, for example, temperature sensitive (ts) and/or cold adapted (ca) vaccine strains. Alternatively, chimeric RSV and vaccine compositions thereof incorporate other mutations specifying desired structural and/or phenotypic characteristics in an infectious chimeric RSV. Such chimeric RSV incorporate desired mutations specified by insertion, deletion, substitution or rearrangement of one or more selected nucleotide sequence(s), gene(s), or gene segment(s) in a chimeric RSV clone. This provides a method for development of novel vaccines against diverse RSV strains by using a common attenuated backbone as a vector to express protective antigens of heterologous strains. The immune system of an individual is stimulated to induce protection against natural RSV infection, preferably in a multivalent manner to achieve protection against multiple RSV strains and/or subgroups.
摘要翻译：嵌合呼吸道合胞病毒（RSV）及其疫苗组合物通过将一个或多个异源基因或基因区段从一个RSV亚组或菌株引入不同亚组或菌株的受体RSV背景中而产生。所得到的嵌合RSV病毒或亚病毒颗粒是感染性和减毒性的，优选通过将指定减毒表型的选定突变引入到嵌合基因组或反基因组中以产生例如温度敏感（ts）和/或冷适应（ca）疫苗株。或者，嵌合RSV及其疫苗组合物掺入在感染性嵌合RSV中指定所需结构和/或表型特征的其它突变。这种嵌合RSV包含通过嵌合RSV克隆中的一个或多个选定的核苷酸序列，基因或基因片段的插入，缺失，取代或重排而指定的所需突变。这提供了通过使用共同的减毒骨架作为表达异源株的保护性抗原的载体来开发针对不同RSV菌株的新型疫苗的方法。刺激个体的免疫系统以诱导针对天然RSV感染的保护，优选以多价方式实现针对多个RSV菌株和/或亚组的保护。

34. 发明授权

US07842798B2 Production of attenuated, human-bovine chimeric respiratory syncytial virus vaccines 失效
标题翻译：减毒，人 - 牛嵌合呼吸道合胞病毒疫苗的生产
公开(公告)号：US07842798B2
公开(公告)日：2010-11-30
申请号：US11203620
申请日：2005-08-11
申请人： Ursula Buchholz , Peter L. Collins , Brian R. Murphy , Stephen S. Whitehead , Christine D. Krempl
发明人： Ursula Buchholz , Peter L. Collins , Brian R. Murphy , Stephen S. Whitehead , Christine D. Krempl
IPC分类号： C07H21/00 , A61K39/155
CPC分类号： C12N7/00 , A61K39/00 , A61K39/12 , A61K39/155 , A61K2039/5254 , A61K2039/5256 , A61K2039/544 , C07K14/005 , C07K2319/00 , C12N15/86 , C12N2760/18522 , C12N2760/18534 , C12N2760/18543 , C12N2760/18561 , C12N2840/203
摘要： Chimeric human-bovine respiratory syncytial virus (RSV) are infectious and attenuated in humans and other mammals and useful in vaccine formulations for eliciting an anti-RSV immune response. Also provided are isolated polynucleotide molecules and vectors incorporating a chimeric RSV genome or antigenome which includes a partial or complete human or bovine RSV “background” genome or antigenome combined or integrated with one or more heterologous gene(s) or genome segment(s) of a different RSV strain. In preferred aspects of the invention, chimeric RSV incorporate a partial or complete bovine RSV background genome or antigenome combined with one or more heterologous gene(s) or genome segment(s) from a human RSV. A variety of additional mutations and nucleotide modifications are provided within the human-bovine chimeric RSV of the invention to yield desired phenotypic and structural effects.
摘要翻译：嵌合人 - 牛呼吸道合胞病毒（RSV）在人类和其他哺乳动物中具有感染性和减毒性，并且可用于引发抗RSV免疫应答的疫苗制剂中。还提供了分离的多核苷酸分子和掺入嵌合RSV基因组或抗原组的载体，其包括部分或完整的人或牛RSV“背景”基因组或与一个或多个异源基因或基因组片段不同的RSV菌株。在本发明的优选方面，嵌合RSV包含与来自人RSV的一个或多个异源基因或基因组片段组合的部分或完整的牛RSV背景基因组或反义基因组。在本发明的人 - 牛嵌合RSV内提供了多种额外的突变和核苷酸修饰，以产生所需的表型和结构效果。

35. 发明授权

US07662397B2 Respiratory syncytial virus vaccines expressing protective antigens from promoter-proximal genes 有权
标题翻译：呼吸道合胞病毒疫苗从启动子近端基因表达保护性抗原
公开(公告)号：US07662397B2
公开(公告)日：2010-02-16
申请号：US11054343
申请日：2005-02-08
申请人： Christine D. Krempl , Peter L. Collins , Brian R. Murphy , Ursula Buchholz , Stephen S. Whitehead
发明人： Christine D. Krempl , Peter L. Collins , Brian R. Murphy , Ursula Buchholz , Stephen S. Whitehead
IPC分类号： A61K39/155 , A61K39/12
CPC分类号： C07K14/005 , A61K39/12 , A61K39/155 , A61K2039/5254 , A61K2039/5256 , A61K2039/543 , C12N7/00 , C12N15/86 , C12N2760/18522 , C12N2760/18534 , C12N2760/18543 , C12N2760/18561 , C12N2760/18622
摘要： Recombinant respiratory syncytial virus (RSV) having the position of genes shifted within the genome or antigenome of the recombinant virus are infectious and attenuated in humans and other mammals. Gene shifted RSV are constructed by insertion, deletion or rearrangement of genes or genome segments within the recombinant genome or antigenome and are useful in vaccine formulations for eliciting an anti-RSV immune response. Also provided are isolated polynucleotide molecules and vectors incorporating a recombinant RSV genome or antigenome wherein a gene or gene segment is shifted to a more promoter-proximal or promoter-distal position within the genome or antigenome compared to a wild type position of the gene in the RSV gene map. Shifting the position of genes in this manner provides for a selected increase or decrease in expression of the gene, depending on the nature and degree of the positional shift.
摘要翻译：重组呼吸道合胞病毒（RSV）具有在重组病毒的基因组或反向基因组内移位的基因位置，在人和其他哺乳动物中具有传染性和减毒性。通过在重组基因组或反向原核基因组内的基因或基因组片段的插入，缺失或重排构建基因转移的RSV，并且可用于引发抗RSV免疫应答的疫苗制剂中。还提供了分离的多核苷酸分子和掺入重组RSV基因组或反基因组的载体，其中与基因组或基因片段中的基因的野生型位置相比，基因或基因片段转移到基因组或反基因组内的更多启动子近端或启动子远端位置 RSV基因图。以这种方式移动基因的位置提供基因的选择性增加或降低，这取决于位置偏移的性质和程度。

36. 发明申请

US20080138861A1 Production of attenuated respiratory syncytial virus vaccines involving modification of M2 ORF2 有权
标题翻译：涉及M2 ORF2修饰的减毒呼吸道合胞病毒疫苗的生产
公开(公告)号：US20080138861A1
公开(公告)日：2008-06-12
申请号：US11011502
申请日：2004-12-13
申请人： Peter L. Collins , Brian R. Murphy , Alison Bermingham
发明人： Peter L. Collins , Brian R. Murphy , Alison Bermingham
IPC分类号： C12N7/01 , C12N15/45 , C12N15/63
CPC分类号： A61K39/155 , A61K39/12 , A61K2039/5254 , A61K2039/543 , A61K2039/544 , C07K14/005 , C12N7/00 , C12N2760/18522 , C12N2760/18534 , C12N2760/18561
摘要： Recombinant respiratory syncytial virus (RSV) are provided in which expression of the second translational open reading frame encoded by the M2 gene (M2ORF2) is reduced or ablated to yield novel RSV vaccine candidates. Expression of M2 ORF2 is reduced or ablated by modifying a recombinant RSV genome or antigenome to incorporate a frame shift mutation, or one or more stop codons in M2 ORF2. Alternatively, M2 ORF2 is deleted in whole or in part to render the M2-2 protein partially or entirely non-functional or to disrupt its expression altogether. M2 ORF2 deletion and knock out mutants possess highly desirable phenotypic characteristics for vaccine development. These changes specify one or more desired phenotypic changes in the resulting virus or subviral particle. Vaccine candidates are generated that show a change in mRNA transcription, genomic or antigenomic RNA replication, viral growth characteristics, viral antigen expression, viral plaque size, and/or a change in cytopathogenicity. In addition, M2-2 knock out or deletion virus exhibits increased levels of synthesis of viral proteins in cell culture, providing an enriched source of viral antigen or protein for purification and use as a noninfectious subunit vaccine.
摘要翻译：提供了重组呼吸道合胞病毒（RSV），其中由M2基因（M2ORF2）编码的第二翻译开放阅读框的表达被减少或消融以产生新的RSV疫苗候选物。 M2 ORF2的表达通过修饰重组RSV基因组或抗原组合来结合帧移位突变或M2 ORF2中的一个或多个终止密码子来减少或消除。或者，M2 ORF2全部或部分缺失，以使M2-2蛋白部分或完全不起作用或完全破坏其表达。 M2 ORF2缺失和敲除突变体对于疫苗开发具有非常需要的表型特征。这些变化指定所得病毒或亚病毒颗粒中的一种或多种期望的表型变化。产生疫苗候选物，其显示mRNA转录，基因组或反基因组RNA复制，病毒生长特征，病毒抗原表达，病毒斑块大小和/或细胞致病性变化的变化。此外，M2-2敲除或缺失病毒在细胞培养物中表现出增加的病毒蛋白质合成水平，提供用于纯化和用作非感染性亚单位疫苗的病毒抗原或蛋白质的丰富来源。

37. 发明授权

US06410023B1 Recombinant parainfluenza virus vaccines attenuated by deletion or ablation of a non-essential gene 有权
标题翻译：重组副流感病毒疫苗通过非必需基因的缺失或消融而减弱
公开(公告)号：US06410023B1
公开(公告)日：2002-06-25
申请号：US09350821
申请日：1999-07-09
申请人： Anna P. Durbin , Peter L. Collins , Brian R. Murphy
发明人： Anna P. Durbin , Peter L. Collins , Brian R. Murphy
IPC分类号： A61K3912
CPC分类号： C12N7/00 , A61K39/00 , A61K2039/522 , C07K14/005 , C12N2760/18622 , C12N2760/18643 , C12N2760/18661
摘要： Recombinant parainfluenza virus (PIV) are provided in which expression of the C, D and/or V translational open reading frame(s) (ORFs) is reduced or ablated to yield novel PIV vaccine candidates. Expression of the C, D and/or V ORF(s) is reduced or ablated by modifying a recombinant PIV genome or antigenome, for example by introduction of a stop codon, by a mutation in an RNA editing site, by a mutation that alters the amino acid specified by an initiation codon, or by a frame shift mutation in the targeted ORF(s). Alternatively, the C, D and/or V ORF(s) is deleted in whole or in part to render the protein(s) encoded thereby partially or entirely non-functional or to disrupt protein expression altogether. C, D and/or V ORF(s) deletion and knock out mutants possess highly desirable phenotypic characteristics for vaccine development. These deletion and knock out mutations changes specify one or more desired phenotypic changes in the resulting virus or subviral particle. Vaccine candidates are generated that show a change in viral growth characteristics, attenuation, plaque size, and/or a change in cytopathogenicity, among other novel phenotypes. A variety of additional mutations and nucleotide modifications are provided within the C, D and/or V ORF(s) deletion or ablation mutant PIV of the invention to yield desired phenotypic and structural effects.
摘要翻译：提供了重组副流感病毒（PIV），其中C，D和/或V翻译开放阅读框（ORF）的表达被降低或消融以产生新型PIV疫苗候选物。 C，D和/或V ORF的表达通过修饰重组PIV基因组或反义基因组，例如通过引入终止密码子，通过RNA编辑位点中的突变，通过改变的突变来降低或消除由起始密码子指定的氨基酸或靶向ORF中的帧位移突变。或者，全部或部分删除C，D和/或V ORF，以使由此编码的蛋白质部分或完全不起作用或完全破坏蛋白质表达。 C，D和/或V ORF（s）缺失和敲除突变体对疫苗开发具有非常需要的表型特征。这些缺失和敲除突变变化指定了所得病毒或亚病毒颗粒中的一种或多种期望的表型变化。产生疫苗候选物，其显示病毒生长特征，衰减，斑块大小和/或细胞致病性变化以及其他新型表型之间的变化。本发明的C，D和/或V ORF缺失或消融突变体PIV中提供了多种其它突变和核苷酸修饰，以产生所需的表型和结构效果。

38. 发明申请

US20190040365A1 RECOMBINANT RESPIRATORY SYNCYTIAL VIRUS STRAINS WITH MUTATIONS IN THE M2-2 ORF PROVIDING A RANGE OF ATTENUATION PHENOTYPES 审中-公开
公开(公告)号：US20190040365A1
公开(公告)日：2019-02-07
申请号：US16061314
申请日：2016-12-12
申请人： Peter L. Collins , Ursula J. Buchholz , Cindy Luongo
发明人： Peter L. Collins , Ursula J. Buchholz , Cindy Luongo
IPC分类号： C12N7/00 , A61K39/155
摘要： Provided herein are novel recombinant respiratory syncytial viruses (RSV) having an attenuated phenotype that contain mutations in the M2-2 open reading frame that interfere with the expression of the M2-2 protein. The M2-2 mutations may be present in combination with mutations at other loci. Using methods described herein, combinations of mutations are provided to achieve desired levels of attenuation. The recombinant RSV strains described here are suitable for use as live-attenuated RSV vaccines. Also provided are polynucleotide sequences of the described viruses, as well as methods for producing and using the viruses.

39. 发明授权

US06790449B2 Methods for producing self-replicating infectious RSV particles comprising recombinant RSV genomes or antigenomes and the N, P, L, and M2 proteins 有权
标题翻译：用于产生包含重组RSV基因组或抗原单核细胞和N，P，L和M2蛋白质的自复制感染性RSV颗粒的方法
公开(公告)号：US06790449B2
公开(公告)日：2004-09-14
申请号：US09847173
申请日：2001-05-03
申请人： Peter L. Collins
发明人： Peter L. Collins
IPC分类号： A61K39155
CPC分类号： C07K14/005 , A61K39/00 , A61K2039/525 , C12N7/00 , C12N15/86 , C12N2760/18521 , C12N2760/18522 , C12N2760/18543
摘要： RNA synthesis by the paramyxovirus respiratory syncytial virus (RSV), a ubiquitous human pathogen, was found to be more complex than previously appreciated for the nonsegmented negative-strand RNA viruses. Intracellular RNA replication of a plasmid-encoded “minigenome” analog of viral genomic RNA was directed by coexpression of the nucleocapsid (N) protein, nucleocapsid phosphoprotein (P), and the large polymerase (L) protein. But, under these conditions, it appeared that the greater part of mRNA synthesis terminated prematurely. However, coexpression of the M2 (ORF1) gene resulted in the efficient production of full-length mRNA. Thus, these results demonstrate that expression of the upstream ORF1, which encoded the previously described 22-kDa M2 protein, was associated with transcription elongation. Accordingly, the claimed invention is directed toward infectious recombinant RSV particles which comprise a recombinant genome or antigenome, as well as the RSV proteins N, P, L, and the RNA polymerase elongation factor M2. This system will permit the introduction of defined changes into infectious RSV which will prove useful in a variety of applications, such as the analysis of RSV molecular biology and pathogenesis, the development of attenuated RSV immunogenic compositions for the preparation of RSV-specific immunological reagents, and the expression of foreign antigens.
摘要翻译：发现通过普遍存在的人类病原体的副粘病毒呼吸道合胞病毒（RSV）的RNA合成比以前对非分段负链RNA病毒所认识的更为复杂。病毒基因组RNA的质粒编码的“小基因组”类似物的细胞内RNA复制通过核衣壳（N）蛋白，核壳蛋白磷蛋白（P）和大聚合酶（L）蛋白的共表达来引导。但是，在这些条件下，mRNA合成的较大部分似乎过早终止。然而，M2（ORF1）基因的共表达导致全长mRNA的有效产生。因此，这些结果表明编码前述的22-kDa M2蛋白的上游ORF1的表达与转录延伸有关。因此，要求保护的发明涉及包含重组基因组或反向原子团以及RSV蛋白N，P，L和RNA聚合酶延伸因子M2的感染性重组RSV颗粒。该系统将允许将感染性RSV引入定义的变化，这将在各种应用中有用，例如RSV分子生物学和发病机理的分析，用于制备RSV特异性免疫试剂的减毒RSV免疫原性组合物的开发，和外源抗原的表达。

40. 发明授权

US5716823A Human respiratory virus preparahons and processess 失效
标题翻译：人类呼吸道病毒预处理和过程
公开(公告)号：US5716823A
公开(公告)日：1998-02-10
申请号：US854783
申请日：1997-05-12
申请人： Gail W. Wertz , Peter L. Collins
发明人： Gail W. Wertz , Peter L. Collins
IPC分类号： A61K38/00 , A61K39/00 , A61K39/155 , C07K14/135 , C12N15/00 , C12N1/12 , C12N1/20 , C12N7/00
CPC分类号： C07K14/005 , A61K39/12 , A61K39/155 , A61K38/00 , A61K39/00 , C12N2760/18522 , C12N2760/18534
摘要： This invention discloses compositions of DNA and proteins that are useful for preparing vaccines against human respiratory syncytial virus �HRSV!. The DNA compositions include structural genes coding for native structural viral proteins and immunogenic fragments of these proteins. Host cells transformed with the above DNA compositions are also disclosed. Vaccines made from the native structural viral proteins or immunogenic fragments are also disclosed as well as methods for protecting humans by inoculation with these vaccines.
摘要翻译：本发明公开了可用于制备针对人呼吸道合胞病毒（HRSV）的疫苗的DNA和蛋白质组合物。 DNA组合物包括编码天然结构病毒蛋白的结构基因和这些蛋白质的免疫原性片段。还公开了用上述DNA组合物转化的宿主细胞。还公开了由天然结构病毒蛋白质或免疫原性片段制备的疫苗以及通过接种这些疫苗来保护人的方法。

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