公开(公告)号：US06713066B1

公开(公告)日：2004-03-30

申请号：US09611829

申请日：2000-07-07

Applicant: Peter L. Collins ,  Brian R. Murphy ,  Alison Bermingham

Inventor： Peter L. Collins ,  Brian R. Murphy ,  Alison Bermingham

IPC: A61K3912

CPC classification number: C07K14/005 ,  A61K39/00 ,  A61K2039/5254 ,  C12N7/00 ,  C12N2760/18522 ,  C12N2760/18543 ,  C12N2760/18561

Abstract: 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.

Abstract translation: 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。

公开(公告)号：US20080138861A1

公开(公告)日：2008-06-12

申请号：US11011502

申请日：2004-12-13

Applicant: Peter L. Collins ,  Brian R. Murphy ,  Alison Bermingham

Inventor： Peter L. Collins ,  Brian R. Murphy ,  Alison Bermingham

IPC: C12N7/01 ,  C12N15/45 ,  C12N15/63

CPC classification number: A61K39/155 ,  A61K39/12 ,  A61K2039/5254 ,  A61K2039/543 ,  A61K2039/544 ,  C07K14/005 ,  C12N7/00 ,  C12N2760/18522 ,  C12N2760/18534 ,  C12N2760/18561

Abstract: 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.

Abstract translation: 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。

公开(公告)号：US07485440B2

公开(公告)日：2009-02-03

申请号：US11011502

申请日：2004-12-13

Applicant: Peter L. Collins ,  Brian R. Murphy ,  Alison Bermingham

Inventor： Peter L. Collins ,  Brian R. Murphy ,  Alison Bermingham

IPC: C12P21/06 ,  A61K39/155

CPC classification number: A61K39/155 ,  A61K39/12 ,  A61K2039/5254 ,  A61K2039/543 ,  A61K2039/544 ,  C07K14/005 ,  C12N7/00 ,  C12N2760/18522 ,  C12N2760/18534 ,  C12N2760/18561

Abstract: 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.

Abstract translation: 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。