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Mutants of clostridium difficile toxin B and methods of use

a technology of clostridium difficile and toxin b, which is applied in the field of mutants of clostridium difficile toxin b and methods of use, can solve the problems of no techniques for blocking intracellular virulence factors, and no techniques which utilize inactive mutants derived from toxins in order

Inactive Publication Date: 2008-05-08
BALLARD JIMMY D +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Currently, there are no techniques for blocking intracellular virulence factors once they have entered the cytosol of cells.
Further, no techniques exist which utilize inactive mutants derived from a toxin in order to inhibit the wild-type toxin at the intracellular cite.

Method used

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  • Mutants of clostridium difficile toxin B and methods of use
  • Mutants of clostridium difficile toxin B and methods of use
  • Mutants of clostridium difficile toxin B and methods of use

Examples

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examples

[0051]During analysis of the TcdB enzymatic domain a set of mutants were identifiable which were unable to modify substrate, yet were capable of blocking TcdB cytopathic effects. Herein are described generation and analyses of these mutants and the demonstration that these proteins are potent intracellular inhibitors of TcdB and block glucosylation of a previously undescribed target. These mutants show, for the first time, that a toxin derivative can be used to effectively block the activity of the native toxin within the cell. This inhibitory activity also suggests a new paradigm for a therapeutic approach to treat toxin-based diseases.

Enzymatic and Cytopathic Activity of Mutants

[0052]As summarized in FIG. 1, 4 deletion and 3 site-directed mutants in the TcdB enzymatic domain were constructed, cloned and isolated from E. coli. The nomenclature for each of these mutants is summarized in panel A of FIG. 1. One site-directed mutant, TcdBW102A wherein the tryptophan at position 102 is ...

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Abstract

An active, or passive vaccine utilizing purified non-toxic mutant TcdB toxins from Clostridium difficile for humans and animals against infections caused by C. difficile and / or C. sordellii. Persons most potentially affected by C. difficile infections include hospitalized patients, infants, and elderly persons. The TcdB toxin mutant of the vaccine preferably lacks the toxicity of a native C. difficile TcdB toxin. A serum comprising antibodies raised to the TcdB toxin mutant is also available for treating humans or animals against C. difficile infections. The serum may be used in a method for conferring passive immunity against C. difficile. Antibodies to the TcdB toxin mutant may be used in diagnostic tests or in treatments to clear TcdB toxin from bodily fluids. The mutant TcdB toxin may be produced by recombinant methods using cDNA encoding the toxin, the cDNA contained for example in a plasmid or host cell.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]The present application is a continuation of U.S. Ser. No. 10 / 463,957, filed Jun. 17, 2003, now U.S. Pat. No. 7,226,597, which claims benefit of U.S. Provisional No. 60 / 389,685, filed Jun. 17, 2002, each of which is explicitly incorporated herein by reference in its entirety.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT[0002]Not Applicable.BACKGROUND[0003]Intracellular bacterial toxins enter cells, modify targets, and in many cases ultimately destroy the targeted cells thereby contributing to the disease process. Currently, there are no techniques for blocking intracellular virulence factors once they have entered the cytosol of cells. Further, no techniques exist which utilize inactive mutants derived from a toxin in order to inhibit the wild-type toxin at the intracellular cite.[0004]Clostridium difficile is the leading cause of hospital acquired diarrhea and pseudomembranous colitis, a multifactorial disease involving...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/08C07K14/00C07K16/18C12N5/06C12N15/11C12N1/20A61P43/00C12P21/04C12N15/00G01N33/00G01N33/53A61K38/00A61K39/00C07K14/33C12N1/21C12N15/31G01N33/569
CPCA61K38/00G01N33/56911C07K14/33A61K39/00A61P43/00
Inventor BALLARD, JIMMY D.SPYRES, LEA M.
Owner BALLARD JIMMY D
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