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Purified vacuolating toxin from Helicobacter pylori and methods to use same
| Details |
Inventors: Cover, Timothy L.; Blaser, Martin J.;
Assignee: Vanderbilt University (Nashville, TN)
Primary Examiner: Housel; James C.
Assistant Examiner: Portner; Ginny Allen
Attorney, Agent or Firm: Needle & Rosenberg
This invention relates to a purified Helicobacter pylori vacuolating toxin and methods to use this toxin to produce protective antibodies against H. pylori infection. Antiserum to this antigen can be used to detect the toxin. Methods to detect anti-toxin antibodies determine the susceptibility of a patient to develop peptic ulcer disease, gastric carcinoma, or other clinical consequences of H. pylori infection. |
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DETAILED DESCRIPTION It is an object of the present invention to provide a substantially pure antigenic composition with vacuolating toxin activity.
It is an object of the present invention to provide a purified antigenic composition that specifically binds antibodies to the toxin.
It is an object of the present invention to provide a clinical diagnostic test for the presence of infection with toxin-producing H.
pylori, and thereby identify patients at risk for peptic ulcer disease or gastric malignancy.
It is an object of the invention to provide a protein vaccine which induces high levels of specific antibodies directed against H.
pylori toxin, and which protects against natural H.
pylori infection in humans.
It is another object of the invention to provide polyclonal or monoclonal antibodies specific for H.
pylori toxin, and methods for their use in detecting the toxin, or for therapeutic purposes.
These and other embodiments are accomplished by providing the antigenic compositions, vaccines, methods, antisera or antibodies, and kits disclosed herein.
In one embodiment of the invention, a purified antigenic composition with vacuolating toxin activity (hereinafter termed CB antigen) is extracted from H.
pylori broth culture supernatant, and has a molecular weight greater than 972,000 daltons (as determined by gel filtration chromatography under nondenaturing conditions), and an apparent molecular weight of 87,000.
+-.
300 daltons when denatured (as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing conditions (SDS-PAGE).
The term CB antigen is defined as the functionally active non-denatured vacuolating toxin; in contrast, the Mr=87,000 protein is a functionally inactive subunit of the CB antigen, which is detected only under denaturing conditions.
The term CB antigen will include antigenic fragments of the holotoxin, whether derived from H.
pylori or synthetically or recombinantly produced.
Proteins having substantial homology to CB antigen or fragments thereof may also be used in accordance with the invention
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