Boots – shoes – and leggings – Orthopedic boot or shoe with corrective element – Arch support
Reexamination Certificate
2002-03-11
2004-04-20
Smith, Lynetter R. F. (Department: 1645)
Boots, shoes, and leggings
Orthopedic boot or shoe with corrective element
Arch support
C424S093480, C424S130100, C424S150100, C424S184100, C424S234100, C424S278100, C435S226000, C435S252100, C435S252330, C435S257400, C435S259000, C436S512000, C436S547000, C436S548000, C514S053000, C514S054000, C514S127000, C530S387100
Reexamination Certificate
active
06722062
ABSTRACT:
FIELD OF THE INVENTION
The present invention is directed to polysaccharides that can be used to induce the production of antibodies to specific strains of enterococcal bacteria. The polysaccharides may be incorporated into a vaccine or used in vitro to assay for the presence of bacteria in a sample of biological fluid.
BACKGROUND OF THE INVENTION
Enterococci are frequently causes of serious infections in newborns and severely immunocompromised patients. Until recently, infections have been adequately controlled using antibiotics. However, drug-resistant bacterial strains are emerging, and infection by strains resistant to all presently available antibiotics may become a serious problem in the near future. It is therefore essential that new methods for controlling enterococcal infections be developed.
The present invention describes a procedure for purifying polysaccharides from the cell membranes of
E. faecalis
. When administered to an appropriate host, these antigens induce the production of high titers of opsonic antibodies that kill both
E. faecalis
and
E. faecium.
SUMMARY OF THE INVENTION
The present invention is based upon the discovery that a polysaccharide antigen can be isolated from
E. faecalis
bacteria and that this antigen can be used to produce antibodies that kill two different species of enterococci,
E. faecalis
and
E. faecium.
E. faecalis
bacteria, preferably bacteria deposited as ATCC number 202159, are sequentially digested with mutanolysin/lysozyme, nucleases (both RNase and DNase) and pronase. The digestion product is then size fractionated and high molecular weight fractions containing the immunogenic polysaccharide are selected. The preferred method for carrying out the size fractionation step is to use a column of Sephacryl S-500™ (Pharmacia, cross-linked allyl dextran/N,N′-methylenebiscraylamide) and to collect fractions corresponding to the void volume.
Structural characterization of the high molecular weight polysaccharide thus obtained has demonstrated that it has a disaccharide, &agr;-D-GlcP-(1→2)-&agr;-D-GlcP, linked to a glycerol teichoic acid backbone via the number 2 carbon of glycerol. There are phosphodiester bonds between the first carbon of the glycerol backbone and the sixth carbon of the glucose disaccharide. Substantially purified high molecular weight polysaccharides having these characteristics are part of the invention. As used herein, the term “substantially purified” refers to polysaccharides that are essentially free from other biological materials. Typically the polysaccharides in such a preparation will constitute at least 80% of the total biological material present with higher percentages being preferred.
Once obtained, the high molecular weight polysaccharides may be incorporated as part of a pharmaceutically acceptable preparation and administered to an animal, e.g., a mouse, rabbit, sheep, goat etc., in order to generate antibodies that react with
E. faecalis
. Alternatively the high molecular weight polysaccharide can be incorporated into a vaccine and administered to people in order to evoke an immune response. As used herein, the term “immune response” means that the administration of vaccine confers upon the recipient protective immunity to subsequent challenge by
E. faecalis
bacteria.
Lower molecular weight polysaccharides may also be generated from the preparations described above. In order to destroy phosphodiester-linked materials, e.g., teichoic acid, the size fractionated material may be treated with hydrogen fluoride. The substantially purified low molecular weight polysaccharides thus obtained may be coupled to a protein or other suitable carrier and then used to generate antibodies or as vaccines in the same manner discussed above.
The high or low molecular weight polysaccharide-containing vaccines may be administered to patients at high risk for enterococci infection. Alternatively, they may be administered to a healthy individual for the purpose of developing antibodies that can be administered to an infected patient. Antibodies developed in a human or other species may also be used in assays to determine whether
E. faecalis
is present in a biological fluid. The present invention includes both the vaccines for immunizing patients as well as the immunization procedure itself.
REFERENCES:
patent: 4242501 (1980-12-01), Cano et al.
patent: 4857512 (1989-08-01), Wagner et al.
patent: 4900677 (1990-02-01), Hewitt
patent: 5292652 (1994-03-01), Dovey et al.
patent: 5472872 (1995-12-01), Mead et al.
patent: 5681736 (1997-10-01), Pace et al.
Wicken, et al., “Structure of Intracellular Teichoic Acids from GroupD. Streptococci,” Biochem J. 87:54-62 (1963).
Huebner Johannes
Madoff Lawrence
Pier Gerald B.
Wang Ying
Fitch Even Tabin & Flannery
Hines Ja'na
Sanzo Michael A.
Smith Lynetter R. F.
The Brigham and Women's Hospital Inc.
LandOfFree
Capsular polysaccharides from enterococci does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Capsular polysaccharides from enterococci, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Capsular polysaccharides from enterococci will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3262299