公开(公告)号：US20090226469A1

公开(公告)日：2009-09-10

申请号：US12279423

申请日：2007-02-16

Applicant: Michael J. Smith ,  Alison D. O'Brien ,  Louise D. Teel ,  Angela R. Melton-Celsa

Inventor： Michael J. Smith ,  Alison D. O'Brien ,  Louise D. Teel ,  Angela R. Melton-Celsa

IPC: A61K39/00 ,  C07K14/00 ,  C07K16/00 ,  C07H21/04 ,  C12N15/63 ,  C12N5/10 ,  C12P21/00 ,  A61P1/12

CPC classification number: C12N15/62 ,  A61K39/00 ,  A61K39/0258 ,  A61K2039/55566 ,  C07K14/245 ,  C07K2319/55 ,  Y02A50/474

Abstract: A chimeric Shiga toxoid according to the invention contains an enzymatically-inactivated StxA subunit and a native StxB subunit. This hybrid Shiga toxoid induces the production of broadly cross-reactive species of antibodies against Shiga toxin following immunization. The StxA subunit is modified so that it is enzymatically inactive. The invention thus encompasses the Shiga toxoid or fragments thereof and the nucleic acid sequence of the Shiga toxoid or fragments thereof. The invention further encompasses the production of a Shiga toxoid, the production of antibodies using the Shiga toxoid and methods of productions, and an immunogenic composition containing the Shiga toxoid.

Abstract translation: 根据本发明的嵌合志贺菌素含有酶促失活的StxA亚基和天然的StxB亚基。 这种混合型志贺毒素在免疫接种后，诱导出针对志贺毒素的广泛交叉反应性抗体的产生。 StxA亚基被修饰，使其酶促无活性。 本发明因此包括志贺毒素或其片段和志贺毒素或其片段的核酸序列。 本发明还包括产生志贺毒素，使用志贺毒素产生抗体和生产方法，以及含有志贺毒素的免疫原性组合物。

公开(公告)号：US06261561B1

公开(公告)日：2001-07-17

申请号：US08840466

申请日：1997-04-18

Applicant: C. Neal Stewart, Jr. ,  Marian L. McKee ,  Alison D. O'Brien ,  Marian R. Wachtel

Inventor： C. Neal Stewart, Jr. ,  Marian L. McKee ,  Alison D. O'Brien ,  Marian R. Wachtel

IPC: A61K39395

CPC classification number: C12N15/8258 ,  A61K38/00 ,  A61K39/025 ,  A61K39/0258 ,  A61K39/0283 ,  A61K47/62 ,  A61K2039/6031 ,  C07K14/24 ,  C07K14/245 ,  C07K16/1228 ,  C07K16/1232 ,  C07K2319/00 ,  C12N15/8242 ,  Y02A50/474 ,  Y02A50/476

Abstract: This invention satisfies needs in the art by providing intimin, the Enterohemorrhagic Escherichia coli (EHEC) adherence protein, alone or as a fusion protein with one or more other antigens, expressed by transgenic plants and the use of those plants as vehicles for stimulating a protective immune response against EHEC and the one or more other antigens. Various plant species are transformed to protect various animal species and also humans against EHEC, against pathogens expressing intimin-like proteins, and against pathogens expressing any of the one or more other antigens to which intimin may be fused. The eae gene encoding intimin, a functional portion thereof, or a recombination that encodes a fusion protein is put under the control of a constitutive plant promoter in a plasmid and the plasmid is introduced into plants by the type of transformation appropriate for the particular plant species. The engineered plants expressing intimin or the intimin fusion protein are then fed to animals and/or humans to elicit the production of antibodies, which protect the animals/humans against EHEC colonization and infection, and against pathogens expressing the one or more other antigens and any cross-reactive antigens. The invention may also be practiced by expressing the intimin or intimin fusion protein in other host organisms such as bacteria, yeast, and fungi.

Abstract translation: 本发明通过以转基因植物单独表达的intimin，肠出血性大肠杆菌（EHEC）粘附蛋白单独或作为与一种或多种其他抗原的融合蛋白，以及使用这些植物作为刺激保护性的载体的方式来满足本领域的需要 针对EHEC和一种或多种其他抗原的免疫应答。 转化各种植物物种以保护各种动物物种和人类免受EHEC，针对表达类胰岛素样蛋白质的病原体，以及表达任何一种或多种其它可能融合的其他抗原的病原体.EEG基因编码intimin，a 将其功能部分或编码融合蛋白的重组置于质粒中的组成型植物启动子的控制下，并通过适合特定植物种类的转化类型将质粒导入植物中。 然后将表达Intimin或Intimin融合蛋白的工程化植物喂养到动物和/或人类以引发抗体的产生，其保护动物/人免受EHEC定植和感染，以及针对表达一种或多种其它抗原的病原体和任何 交叉反应性抗原。 本发明还可以通过在其它宿主生物例如细菌，酵母和真菌中表达intimin或intimin融合蛋白来实施。

公开(公告)号：US06406885B1

公开(公告)日：2002-06-18

申请号：US09696188

申请日：2000-10-26

Applicant: C. Neal Stewart, Jr. ,  Marian L. McKee ,  Alison D. O'Brien ,  Marian R. Wachtel

Inventor： C. Neal Stewart, Jr. ,  Marian L. McKee ,  Alison D. O'Brien ,  Marian R. Wachtel

IPC: C12P2106

CPC classification number: C12N15/8258 ,  A61K38/00 ,  A61K39/025 ,  A61K39/0258 ,  A61K39/0283 ,  A61K47/62 ,  A61K2039/6031 ,  C07K14/24 ,  C07K14/245 ,  C07K16/1228 ,  C07K16/1232 ,  C07K2319/00 ,  C12N15/8242 ,  Y02A50/474 ,  Y02A50/476

Abstract: This invention satisfies needs in the art by providing intimin, the Enterohemorrhagic Escherichia coli (EHEC) adherence protein, alone or as a fusion protein with one or more other antigens, expressed by transgenic plants and the use of those plants as vehicles for stimulating a protective immune response against EHEC and the one or more other antigens. Various plant species are transformed to protect various animal species and also humans against EHEC, against pathogens expressing intimin-like proteins, and against pathogens expressing any of the one or more other antigens to which intimin may be fused. The eae gene encoding intimin, a functional portion thereof, or a recombination that encodes a fusion protein is put under the control of a constitutive plant promoter in a plasmid and the plasmid is introduced into plants by the type of transformation appropriate for the particular plant species. The engineered plants expressing intimin or the intimin fusion protein are then fed to animals and/or humans to elicit the production of antibodies, which protect the animals/humans against EHEC colonization and infection, and against pathogens expressing the one or more other antigens and any cross-reactive antigens. The invention may also be practiced by expressing the intimin or intimin fusion protein in other host organisms such as bacteria, yeast, and fungi.

Abstract translation: 本发明通过以转基因植物单独表达的intimin，肠出血性大肠杆菌（EHEC）粘附蛋白单独或作为与一种或多种其他抗原的融合蛋白，以及使用这些植物作为刺激保护性的载体的方式来满足本领域的需要 针对EHEC和一种或多种其他抗原的免疫应答。 转化各种植物物种以保护各种动物物种和人类免受EHEC，针对表达类胰岛素样蛋白质的病原体，以及表达任何一种或多种其它可能融合的其他抗原的病原体.EEG基因编码intimin，a 将其功能部分或编码融合蛋白的重组置于质粒中的组成型植物启动子的控制下，并通过适合特定植物种类的转化类型将质粒导入植物中。 然后将表达Intimin或Intimin融合蛋白的工程化植物喂养到动物和/或人类以引发抗体的产生，其保护动物/人免受EHEC定植和感染，以及针对表达一种或多种其它抗原的病原体和任何 交叉反应性抗原。 本发明还可以通过在其它宿主生物例如细菌，酵母和真菌中表达intimin或intimin融合蛋白来实施。

公开(公告)号：US08846058B2

公开(公告)日：2014-09-30

申请号：US12279423

申请日：2007-02-16

Applicant: Michael J. Smith ,  Alison D. O'Brien ,  Louise D. Teel ,  Angela R. Melton-Celsa

Inventor： Michael J. Smith ,  Alison D. O'Brien ,  Louise D. Teel ,  Angela R. Melton-Celsa

IPC: A61K39/02 ,  C07K14/245 ,  A61K39/108 ,  C12N15/62 ,  A61K39/00

CPC classification number: C12N15/62 ,  A61K39/00 ,  A61K39/0258 ,  A61K2039/55566 ,  C07K14/245 ,  C07K2319/55 ,  Y02A50/474

Abstract: A chimeric Shiga toxoid according to the invention contains an enzymatically-inactivated StxA subunit and a native StxB subunit. This hybrid Shiga toxoid induces the production of broadly cross-reactive species of antibodies against Shiga toxin following immunization. The StxA subunit is modified so that it is enzymatically inactive. The invention thus encompasses the Shiga toxoid or fragments thereof and the nucleic acid sequence of the Shiga toxoid or fragments thereof. The invention further encompasses the production of a Shiga toxoid, the production of antibodies using the Shiga toxoid and methods of productions, and an immunogenic composition containing the Shiga toxoid.

Abstract translation: 根据本发明的嵌合志贺菌素含有酶促失活的StxA亚基和天然的StxB亚基。 这种混合型志贺毒素在免疫接种后，诱导出针对志贺毒素的广泛交叉反应性抗体的产生。 StxA亚基被修饰，使其酶促无活性。 本发明因此包括志贺毒素或其片段和志贺毒素或其片段的核酸序列。 本发明还包括产生志贺毒素，使用志贺毒素产生抗体和生产方法，以及含有志贺毒素的免疫原性组合物。

公开(公告)号：US20100189715A1

公开(公告)日：2010-07-29

申请号：US12536940

申请日：2009-08-06

Applicant: Jeffrey R. Stinson ,  Hing Wong ,  Alison D. O'Brien ,  Clare K. Schmitt ,  Angela Melton-Celsa

Inventor： Jeffrey R. Stinson ,  Hing Wong ,  Alison D. O'Brien ,  Clare K. Schmitt ,  Angela Melton-Celsa

IPC: A61K39/395 ,  A61P31/04

CPC classification number: C07K16/1232 ,  A61K38/00 ,  C07K2317/24 ,  C07K2319/00

Abstract: The present invention describes the preparation and use of biologically and immunologically active humanized monoclonal antibodies to Shiga toxin, a toxin associated with HC and the potentially life-threatening sequela HUS transmitted by strains of pathogenic bacteria. The present invention describes how these humanized antibodies may be used in the treatment or prevention of Shiga toxin induced diseases. One aspect of the invention is the humanized monoclonal antibody which binds Shiga toxin where the constant regions are IgG1-kappa and the variable regions are murine in origin. Yet another aspect of the invention is expression vectors and host cells transformed with such vectors which express the humanized monoclonal antibodies of the present invention.

Abstract translation: 本发明描述了生物学和免疫学活性的人源化单克隆抗体与志贺毒素（一种与HC相关的毒素）和潜在的危及生命的后遗症HUS由病原菌菌株传播的制备和应用。 本发明描述了这些人源化抗体如何用于治疗或预防志贺毒素诱导的疾病。 本发明的一个方面是结合志贺毒素的人源化单克隆抗体，其中恒定区是IgG1-κ，可变区是起源的鼠。 本发明的另一方面是用表达本发明的人源化单克隆抗体的载体转化的表达载体和宿主细胞。

公开(公告)号：US06942861B2

公开(公告)日：2005-09-13

申请号：US08837459

申请日：1997-04-18

Applicant: Marian L. McKee ,  Alison D. O'Brien ,  Marian R. Wachtel

Inventor： Marian L. McKee ,  Alison D. O'Brien ,  Marian R. Wachtel

IPC: C12N15/09 ,  A61K31/00 ,  A61K38/00 ,  A61K39/00 ,  A61K39/02 ,  A61K39/112 ,  A61K39/395 ,  A61K47/48 ,  A61P37/00 ,  C07K14/24 ,  C07K14/245 ,  C07K16/12 ,  C12N15/00 ,  C12N15/31 ,  C12N15/82 ,  C12P21/02 ,  C12P21/08 ,  C12R1/19

CPC classification number: C12N15/8258 ,  A61K38/00 ,  A61K39/025 ,  A61K39/0283 ,  A61K47/62 ,  A61K2039/6031 ,  C07K14/24 ,  C07K14/245 ,  C07K16/1228 ,  C07K16/1232 ,  C07K2319/00 ,  Y02A50/476 ,  Y02A50/484

Abstract: The present invention describes the isolation and purification of histidine-tagged functional portions of intimin (his-tagged intimin or his-intimin), a protein associated with the ability of certain strains of pathogenic bacteria to adhere to epithelial cells. The invention further describes the use of intimin as an antigen to promote a protective immune response. In addition, the invention describes the combination of intimin with one or more other antigens and administration of the combination to promote a protective immune response against intimin and the one or more antigens.One aspect of the invention is the administration of intimin to target specific epithelial cells to promote a protective immune response to intimin proteins. Additional aspects of the invention include the use of intimin or intimin combined with one or more antigens and administration of the combination to target gastrointestinal mucosa and stimulate an immune response. Additionally, the invention describes administration of the combination of intimin combined with drugs, to provide a means for targeted delivery of drugs to specific epithelial cells. Other aspects of the invention include the production of antibodies directed against his-intimin and methods of using such antibodies to provide passive immune protection, and in an assay system.

Abstract translation: 本发明描述了分泌和纯化组蛋白标记的功能性部分的intimin（his-tagged intimin或his-intimin），一种与某些病原菌菌株粘附于上皮细胞的能力相关的蛋白质。 本发明进一步描述了作为抗原的肠溶蛋白作为促进保护性免疫应答的用途。 此外，本发明描述了肠溶蛋白与一种或多种其它抗原的组合以及组合的施用，以促进针对胰岛素和一种或多种抗原的保护性免疫应答。 本发明的一个方面是给予内膜以靶向特异性上皮细胞以促进对肠溶蛋白的保护性免疫应答。 本发明的另外的方面包括使用与一种或多种抗原组合的肠溶蛋白或肠溶蛋白和给予组合以靶向胃肠粘膜并刺激免疫应答。 另外，本发明描述了与药物组合的联合应用，以提供用于将药物靶向递送到特定上皮细胞的手段。 本发明的其它方面包括产生针对他的肠溶蛋白的抗体以及使用这些抗体提供被动免疫保护的方法，以及在测定系统中。

公开(公告)号：US20100166753A1

公开(公告)日：2010-07-01

申请号：US12647845

申请日：2009-12-28

Applicant: Jeffrey R. Stinson ,  Hing Wong ,  Alison D. O'Brien ,  Clare K. Schmitt ,  Angela Melton-Celsa

Inventor： Jeffrey R. Stinson ,  Hing Wong ,  Alison D. O'Brien ,  Clare K. Schmitt ,  Angela Melton-Celsa

IPC: A61K39/40 ,  A61P13/12 ,  A61P19/02 ,  A61P37/02 ,  A61P1/02

CPC classification number: C07K16/1232 ,  A61K38/00 ,  C07K2317/24 ,  C07K2319/00

Abstract: The present invention describes the preparation and use of biologically and immunologically active humanized monoclonal antibodies to Shiga toxin, a toxin associated with HC and the potentially life-threatening sequela HUS transmitted by strains of pathogenic bacteria. The present invention describes how these humanized antibodies may be used in the treatment or prevention of Shiga toxin induced diseases. One aspect of the invention is the humanized monoclonal antibody which binds Shiga toxin where the constant regions are IgG1-kappa and the variable regions are murine in origin. Yet another aspect of the invention is expression vectors and host cells transformed with such vectors which express the humanized monoclonal antibodies of the present invention.

公开(公告)号：US06881411B2

公开(公告)日：2005-04-19

申请号：US10150058

申请日：2002-05-20

Applicant: C. Neal Stewart, Jr. ,  Marian L. McKee ,  Alison D. O'Brien ,  Marian R. Wachtel

Inventor： C. Neal Stewart, Jr. ,  Marian L. McKee ,  Alison D. O'Brien ,  Marian R. Wachtel

IPC: C12N15/09 ,  A61H5/00 ,  A61K31/00 ,  A61K38/00 ,  A61K39/02 ,  A61K39/108 ,  A61K39/112 ,  A61K47/48 ,  A61P31/00 ,  A61P31/04 ,  A61P37/04 ,  C07K14/24 ,  C07K14/245 ,  C07K14/415 ,  C07K16/12 ,  C12N5/10 ,  C12N15/31 ,  C12N15/82 ,  C12P21/06

CPC classification number: C12N15/8258 ,  A61K38/00 ,  A61K39/025 ,  A61K39/0258 ,  A61K39/0283 ,  A61K47/62 ,  A61K2039/6031 ,  C07K14/24 ,  C07K14/245 ,  C07K16/1228 ,  C07K16/1232 ,  C07K2319/00 ,  C12N15/8242 ,  Y02A50/474 ,  Y02A50/476

Abstract: This invention satisfies needs in the art by providing intimin, the Enterohemorrhagic Escherichia coli (EHEC) adherence protein, alone or as a fusion protein with one or more other antigens, expressed by transgenic plants and the use of those plants as vehicles for stimulating a protective immune response against EHEC and the one or more other antigens. Various plant species are transformed to protect various animal species and also humans against EHEC, against pathogens expressing intimin-like proteins, and against pathogens expressing any of the one or more other antigens to which intimin may be fused.The eae gene encoding intimin, a functional portion thereof, or a recombination that encodes a fusion protein is put under the control of a constitutive plant promoter in a plasmid and the plasmid is introduced into plants by the type of transformation appropriate for the particular plant species. The engineered plants expressing intimin or the intimin fusion protein are then fed to animals and/or humans to elicit the production of antibodies, which protect the animals/humans against EHEC colonization and infection, and against pathogens expressing the one or more other antigens and any cross-reactive antigens. The invention may also be practiced by expressing the intimin or intimin fusion protein in other host organisms such as bacteria, yeast, and fungi.

Abstract translation: 本发明通过以转基因植物单独表达的intimin，肠出血性大肠杆菌（EHEC）粘附蛋白单独或作为与一种或多种其他抗原的融合蛋白，以及使用这些植物作为刺激保护性的载体的方式来满足本领域的需要 针对EHEC和一种或多种其他抗原的免疫应答。 转化各种植物物种以保护各种动物物种和人类免受EHEC，针对表达类胰岛素样蛋白质的病原体，以及针对表达任何一种或多种其它可能融合的其他抗原的病原体。 将编码融合蛋白的蛋白质的eae基因，其功能部分或编码融合蛋白的重组置于质粒中的组成型植物启动子的控制下，并通过适合特定植物种类的转化类型将质粒导入植物中 。 然后将表达Intimin或Intimin融合蛋白的工程化植物喂养到动物和/或人类以引发抗体的产生，其保护动物/人免受EHEC定植和感染，以及针对表达一种或多种其它抗原的病原体和任何 交叉反应性抗原。 本发明还可以通过在其它宿主生物例如细菌，酵母和真菌中表达intimin或intimin融合蛋白来实施。