US Government As Represented by the Secretary of the Army

The United States of America As Represented by the Secretary of

Frederick J Cassels

Edgar C Boedeker

Robert H Reid

Marcia Kay Wolf

Daniel L Jarboe

Thomas R Tice

Andrew Lees

Richard F Schuman

Jean A Setterstrom

John E Van Hamont

F Donald Roberts

Curt Thies

Charles E McQueen

Elliot Jacob

William R Brown

Phil Friden

Jack London

Ramasubbu Jeyanthi

Gregory M Glenn

Lewis K Pannell

James F Wood

Hyoik Ryu

Lawrence Loomis-Price

Brian A Bell

Disclosed herein are antigens that stimulate protective antibodies against enterotoxigenic. Also disclosed herein are proteins encoded by cssA and cssB genes as well as constructs containing the genes and methods of using thereof.

Transformed bacteria producing CS6 antigens as vaccines

This invention is directed to oral-intestinal vaccines and their use against diseases caused by enteropathogenic organisms using antigens encapsulated within biodegradable-biocompatible microspheres.

Oral-intestinal vaccines against diseases caused by enteropathic organisms using antigens encapsulated within biodegradable-biocompatible microspheres

A monoclonal antibody to a consensus peptide of the formula: The monoclonal antibody of the invention binds exclusively to the sequence SAVALTYS (SEQ ID NO:2) and has use as a diagnostic and for prophylaxis against illness arising fromwhich produce the CS4-CFA/I family of proteins and for treatment of disease arising therefrom.

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A vaccine delivered by transcutaneous immunization provides an effective treatment against infections by pathogens such as, for example, enterotoxigenic(ETEC) and/or for symptoms of diarrheal disease caused thereby. For example, one, two, three, four, five or more antigens derived from ETEC and capable of inducing an antigen-specific immune response (e.g., toxins, colonization or virulence factors) and one or more optional adjuvant (e.g., whole bacterial ADP-ribosylating exotoxins, B subunits or toxoids thereof, detoxified mutants and derivatives thereof) are used to manufacture vaccines or to induce systemic and/or mucosal immunity.

Vaccine for transcutaneous immunization

This invention relates to amino acid sequences from within a consensus peptide of the formula: Eight mer peptides from within the consensus peptide were tested against an antibody raised to the consensus peptide. Studies relating to antibody raised to denatured proteins from the natural organisms producing the family of proteins were also useful and showed particular value of some sequences. A sequence of the formula ASVDPTIDLLQA (SEQ ID NO: 2) was identified thereby. An enlarge sequence of the formula TVTASVDPTIDLLQAD (SEQ ID NO: 3) is also especially interesting as are intermediate sequences such as sequences VTASVDPTIDLLQAD (SEQ ID NO: 4), TASVDPTIDLLQAD (SEQ ID NO: 5), and TASVDPTIDLLQA (SEQ ID NO: 6) as being binding sites for antibodies raised to the denatured proteins.

Peptides responsive to antibodies against consensus peptide of the CS4-CFA/I family proteins

Disclosed herein are methods for making large amounts of highly pure colonization factors. The methods of the present invention differ from prior art methods in that host cells which express the colonization factor of interest are cultured in media comprising more than about 50 &#x3bc;g/l of an antibiotic, the media is centrifuged and then filtered with a 0.2 &#x3bc;m filter tangential flow cartridge and a 300,000 MW cut-off filter, and a divalent cation is added. As disclosed herein the colonization factors made by the method of the present invention may be used in pharmaceutical compositions and methods for treating or preventing enterotoxigenicinfections.

A monoclonal antibody to a consensus peptide of the formula: The monoclonal antibody of the invention binds exclusively to the sequence SAVALTYS (SEQ ID NO. 2) and has use as a diagnostic and for prophylaxis against illness arising fromwhich produce the CS4-CFA/I family of proteins and for treatment of disease arising therefrom.

Monoclonal antibody which agglutinates E. coli having the CS4-CFA/I family protein

Disclosed herein are RBC preparations and methods of making and using thereof for treating, preventing or inhibiting ETEC infections. In particular, bovine red blood cell (RBC) preparations are shown to reduce enterotoxigenic(ETEC) adhesion when administered orally. Also disclosed are kits comprising RBC preparations for the treatment of ETEC infections.

Method for treating, preventing, or inhibiting enterotoxigenic Escherichia coli infections with bovine red blood cells

Novel burst-free, sustained release biocompatible and biodegrable microcapsules which can be programmed to release their active core for variable durations ranging from 1-100 days in an aqueous physiological environment. The microcapsules are comprised of a core of polypeptide or other biologically active agent encapsulated in a matrix of poly(lactide/glycolide) copolymer, which may contain a pharmaceutically-acceptable adjuvant, as a blend of upcapped free carboxyl end group, and end-capped forms ranging in ratios from 100/0 to 1/99.

Therapeutic treatment and prevention of infections with a bioactive materials encapsulated within a biodegradable-biocompatible polymeric matrix

Disclosed herein are methods for identifying at least one bacterial colonization factor of enterotoxigenic which comprise the following steps in the following order: 1) obtaining the colonization factor; 2) solubilizing the colonization factor by dissolving the colonization factor in 1,1,1,3,3,3-hexafluoro-2-propanol; 3) adding a solution of volatile acid to the solubilized colonization factor of step 2 to obtain a product; 4) subjecting the product of step 3 to mass spectrometry to determine the mass of the colonization factor; and 5) comparing the mass determined in step 4 with the mass of at least one known colonization factor.

Mass spectrometry of colonization factors

Novel burst-free, sustained release biocompatible and biodegrable microcapsules which can be programmed to release their active core for variable durations ranging from 1-100 days in an aqueous physiological environment. The microcapsules are comprised of a core of polypeptide or other biologically active agent encapsulated in a matrix of poly(lactide/glycolide) copolymer, which may contain a pharmaceutically-acceptable adjuvant, as a blend of upcapped free carboxyl end group and end-capped forms ranging in ratios from 100/0 to 1/99.

A consensus poptide of 36 amino acids has been designed which acts as an immunogen raising antibodies against the proteins of all members of the E. coli family CS4-CFA/1. While the N-terminus of members of this family of organisms shows a high degree of identity, the remainder of the sequence of the proteins shows much less homology across the strains. The region of the protein represented in the subunit encompasses known linear B- and T-cell epitopes of CFA/I. The consensus peptide has a high level of homology to strains bearing six different colonization factors. The consensus peptide is of the formula:

Method of raising antibodies against E. coli of the family CS4-CFA/I

Production of proteins in bacteria containing DNA sequences encoding proteins under the control of a temperature-regulated promotor is improved by growing the organisms at temperatures of less than 35.degree. C. until the late logarithmic phase. Thereafter the temperature may be raised to 36.degree. C. to 39.degree. C. Antigens produced by the method of the invention may be used as vaccines, as means for measuring efficacy of vaccines, as probes to detect antigens from clinical samples and for biochemical characterization of antigens.

Methods for production of antigens under control of

A purified oligosaccharide consisting of a hexasaccharide with a native molecular weight of 959 which can be isolated from the cell walls of Streptococcus oralis. The oligosaccharide is able to significantly block the interaction between the human oral plaque bacteria Streptococcus oralis H1 and Capnocytophaga ochracea ATCC 33596. This purified oligosaccharide contains saccharide components found to inhibit many know interactions between plaque bacteria and may be effective in prevention, inhibition and reversal of dental plaque deposits. The oligosaccharide may be applied effectively when incorporated in toothpastes, mouth wash, etc.

Growing community of inventors

Laurel, MD, United States of America

Frederick J Cassels