The Henry M. Jackson Foundation for the Advancement of Military Medicine Inc.

Sunol Molecular Corporation

Alison D O'Brien

Angela Ruth Melton-Celsa

Marian L McKee

Michael Smith

Marian R Wachtel

C Neal Stewart

James Sinclair

Hing C Wong

Jeffrey R Stinson

Liyanage Parakrama Perera

Susanne Ward Lindgren

Nancy A Strockbine

Louise D Teel

Clare K Schmitt

Michael J Smith

The invention is based on the discovery of the epitope in the Stx2 protein for the 11 E1O antibody. The invention features compositions containing non-full length Stx2 polypeptides that include the 11 E1O monoclonal antibody epitope. The invention also features methods of producing anti-Stx2 antibodies specific for the 11 E1O epitope of the Stx2 protein. Additionally, the invention features methods for treating a subject having, or at risk of developing, a Shiga toxin associated disease (e.g., hemolytic uremia syndrome and diseases associated withandinfection) with a polypeptide that includes the 11 E1O epitope or with an anti-Stx2 antibody developed using the methods of the invention. Furthermore, the invention features the detection of Stx2 in a sample using the antibodies developed using the methods of the invention.

Methods and compositions based on Shiga toxin type 2 protein

Methods and compositions based on shiga toxin type 2 protein

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.

Shiga toxoid chimeric proteins

The invention is based on the discovery of the epitope in the Stx1 protein for the 13C4 antibody. The invention features non-full length Stx1 polypeptides that include the epitope for the 13C4 monoclonal antibody epitope. The invention also features methods of producing anti-Stx1 antibodies specific for the 13C4 epitope of the Stx1 protein. Additionally, the invention features methods for treating a subject having, or at risk of developing, a Shiga toxin associated disease (e.g., hemolytic uremia syndrome and diseases associated withandinfection) with a polypeptide that includes the 13C4 epitope or with an anti-Stx1 antibody developed using the methods of the invention. Furthermore, the invention features the detection of Stx1 in a sample using the antibodies developed using the methods of the invention.

Methods and compositions based on Shiga toxin type 1 protein

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.

Histidine-tagged intimin and methods of using intimin to stimulate an immune response and as an antigen carrier with targeting capability

This invention satisfies needs in the art by providing intimin, the Enterohemorrhagic(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.

Method of stimulating an immune response by administration of host organisms that express intimin alone or as a fusion protein with one or more other antigens

Plants and plant cells expressing histidine tagged intimin

A rapid, sensitive, non-radioactive diagnostic kit for the direct detection of both Shiga-like toxin, type I, and Shiga-like toxin, type II, produced by enterohemorrhagic Escherichia Coli in food and clinical samples. This diagnostic kit is comprised of a monoclonal antibody capable of detecting Shiga-like toxin, type I, and a monoclonal antibody capable of detecting Shiga-like toxin, type II, together with a chemiluminescing detection reagent with a sensitivity enhancer.

Detection of shiga-like toxins of enterohemoragic Escherichia coli

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Bethesda, MD, United States of America

Alison D O'Brien