WO1998012562A1 - Adhesines obtenues d'heliobacter pylori et leurs utilisations therapeutiques et diagnostiques - Google Patents

Adhesines obtenues d'heliobacter pylori et leurs utilisations therapeutiques et diagnostiques Download PDF

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Publication number
WO1998012562A1
WO1998012562A1 PCT/GB1997/002554 GB9702554W WO9812562A1 WO 1998012562 A1 WO1998012562 A1 WO 1998012562A1 GB 9702554 W GB9702554 W GB 9702554W WO 9812562 A1 WO9812562 A1 WO 9812562A1
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WO
WIPO (PCT)
Prior art keywords
pylori
adhesin
subject
kit
adhesive
Prior art date
Application number
PCT/GB1997/002554
Other languages
English (en)
Inventor
Bow Ho
Original Assignee
Cortecs International Limited
Chapman, Paul, William
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from GBGB9619694.4A external-priority patent/GB9619694D0/en
Priority claimed from GBGB9622846.5A external-priority patent/GB9622846D0/en
Application filed by Cortecs International Limited, Chapman, Paul, William filed Critical Cortecs International Limited
Priority to AU43116/97A priority Critical patent/AU4311697A/en
Publication of WO1998012562A1 publication Critical patent/WO1998012562A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/205—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Campylobacter (G)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies

Definitions

  • the present invention relates to novel methods of diagnosing H. pylori infection in a subject.
  • the invention relates to methods for distinguishing between different disease states caused by H. pylori .
  • the invention relates to the use of adhesin proteins in the production of vaccines.
  • H. pylori is a Gram negative bacteria that has been strongly implicated in chronic active gastritis and peptic ulcer disease (Marshall et al , Medical Journal of Aus tralia , 142:439-444 (1985); Buck, G.E., Journal of clinical Microbiology, 3:1-12 (1990)). More recently, it has also been implicated in the development of gastric cancer .
  • H. pylori has been shown to invoke both a systemic and a local immune response (Jones et al , J. Clin . Pathol . , 37:1002-6 (1984); Kaldor et al , Lancet, 1:921 (1985); Rathbone et al , Gu t, 27:642-7 (1986)), and this immune response has been used as a basis for the detection/ diagnosis of H. pylori .
  • HELISALTM is sold as a rapid point of care diagnostic test device, which relies on detection of antibodies against H. pylori found in a subject's blood or saliva.
  • H. pylori has been implicated in a range of gastric disease conditions.
  • simple detection of the presence of the bacteria does not provide a complete picture of the disease state of the subject. It would therefore be preferable to be able to distinguish between particular gastric disease states on the basis of the immune response found in a subject.
  • the importance of bacterial adhesion in infectivity has been widely accepted since it was demonstrated that infectivity generally paralleled adhering ability in bacteria (Beachey, E.H., J " . Infect . Dis .
  • H. pylori possesses adhesive proteins that bind to membrane enriched fraction (MEF) of KATO III cells (Ho, B. and Jiang, B., Euro . J. Gas troen terol . & Hepa tol . , 7(2): 121-124 (1995)). These adhesive proteins (adhesins) can be used to detect anti- H. pylori adhesive protein antibodies.
  • H. pylori adhesive proteins can be indicative of that subject's gastric disease state, and can even be used to distinguish between different gastric conditions .
  • the present invention provides a method for the detection of H. pylori in a subject which comprises the step of contacting at least one H. pylori adhesive protein, or an antibody binding portion thereof, with a biological sample obtained from the subject.
  • the method can be used to diagnose H. pylori infection in a subject.
  • the method of the invention will determine whether antibodies to the at least one adhesive protein (adhesin) , or an antibody binding part thereof, are present in the sample.
  • the adhesive protein is an adhesin.
  • a whole adhesive protein for example an adhesin, or a part thereof which is capable of binding to one or more anti- adhesive protein antibodies.
  • a whole adhesive protein for example an adhesin
  • a part thereof which is capable of binding to one or more anti- adhesive protein antibodies.
  • an epitope containing part of such a protein for example, one can use an epitope containing part of such a protein.
  • antigens or parts thereof can be used in this method.
  • a mixture could include a range of antigenic adhesive proteins as well as other antigenic proteins derived from H. pylori .
  • antigenic proteins include those disclosed in WO-A-93/22682 and in WO-A-96/25430.
  • Bio sample refers to a sample of tissue or, more particularly, a sample of a bodily fluid, eg a sample of blood or saliva.
  • a sample of a bodily fluid eg a sample of blood or saliva.
  • the term is also intended to refer to samples which have been manipulated in some way prior to testing. Examples of such manipulation include separation of red blood cells from serum and filtration and/or pH adjustment of a saliva sample.
  • the method preferably includes detection of IgA antibodies to at least one adhesive protein.
  • the test sensitivity should be increased and enable detection of some individuals who, although exhibiting no general antibody response, should in fact be classified as positive for infection.
  • H. pylori adhesive proteins examples include the known 26 kDa adhesin protein which is antigenic and has the following N-terminal sequence:
  • This novel adhesin protein has been identified from the TIGR H. pylori genome database as the protein superoxide dismutase (SOD) and forms a second aspect of the invention.
  • the protein is provided in an isolated and purified form.
  • the present invention provides a method of assessing the gastric disease state of a subject which comprises the step of determining whether antibodies to at least one H. pylori adhesive protein, for example an adhesin, or an antigenic part thereof, are present in a biological sample obtained from the subject.
  • H. pylori adhesive protein for example an adhesin, or an antigenic part thereof
  • the method includes quantitation of the general H. pylori response and the antibody response to adhesive proteins and comparing the two. Where the anti-adhesive protein response is statistically lower it is possible to classify the subject as suffering from Non Ulcer Dyspepsia (NUD) or Gastritis. However, where the general H. pylori antibody response is statistically lower than the anti-adhesive protein antibody response, this may indicate classification of the subject as suffering from gastric cancer. Where there appears to be no staistical difference between the responses the subject can be classified as suffering from Gastric or Duodenal ulcer. This preliminary classification of a subject's gastric disease state, in combination with other diagnostic means, will enable more rapid and effective treatment of the condition.
  • NUD Non Ulcer Dyspepsia
  • the present invention provides the use of at least one adhesive protein, for example an adhesin, from H. pylori in the detection of H. pylori or in the diagnosis of H. pylori infection.
  • the antigen can be used alone or, more usually, as part of an antigen composition comprising other antigenic proteins from H. pylori .
  • the diagnostic methods of the present invention will be carried out using a test device or test kit, e.g. that used in the HE ISALTM test.
  • the present invention provides a kit for use in the diagnosis of H. pylori infection which comprises at least one adhesive protein, for example an adhesin, from H. pylori .
  • the kit of the invention will also include one or more other antigenic proteins from H. pylori , either other adhesive proteins or other antigens such as those mentioned above.
  • the kit will further comprise one or other components enabling detection of antibodies in accordance with the methods descibed herein.
  • the present invention provides a kit for use in assessing the gastric disease state of a subject which comprises at least one adhesive protein, for example anadhesin, or an antigenic portion thereof, from H. pylori .
  • a kit for use in assessing the gastric disease state of a subject which comprises at least one adhesive protein, for example anadhesin, or an antigenic portion thereof, from H. pylori .
  • such a kit can also include one or more additional antigens from H. pylori .
  • the kit can also include one or more components enabling detection of anti-adhesive protein antibodies and/or one or more components allowing quantitation of the general anti -H .pylori antibody response and the anti- adhesive protein antibody response and their comparison and/or one or more components allowing determination of the relative anti -adhesive protein IgA, IgG and IgM response of the subject.
  • kits of the invention can suitably be provided in the form of a diagnostic device.
  • adhesin antigens particularly the novel antigen disclosed herein, can also be used to produce a vaccine against H. pylori .
  • the present invention provides the use of an adhesive protein, preferably an adhesin, in the manufacture of a vaccine, either for prophylaxis or treatment of H. pylori infections.
  • the vaccine will incorporate the novel adhesin protein disclosed herein.
  • Vaccines incorporating the novel adhesin protein disclosed herein form another aspect of the present invention. Methods of vaccinating a subject using such vaccines are also included within the scope of the invention.
  • NUD NUD
  • gastritis gastric ulcer
  • GU gastric ulcer
  • DU duodenal ulcer
  • GO gastric cancer
  • ELISA for detection of H. pylori infection
  • human sera samples were assayed for the presence of H. pylori IgG, IgM and IgA antibodies by an ELISA method (Khin, M.M., and Ho, B., Biomedical Let ters, 50:71-78 (1994)).
  • ELISA method Khin, M.M., and Ho, B., Biomedical Let ters, 50:71-78 (1994)
  • each plate was tested alongside a set of two control sera that had to meet stringent criteria for the test.
  • the first test control was pooled positive sera diluted from 1:100 in doubling dilutions to 1:3200. This control was used mainly to calculate regression values for the ELISA assays and to analyse the ELISA results.
  • the second was a pooled negative sera at a dilution of 1:100 in triplicate.
  • the ELISA results were calculated using Lotus 123 (version 3.4) in order to calculate regression values for semi-quantitative analysis. Test results were only accepted when the regression value was ⁇ 0.9.
  • Adhesive proteins in cell free extract of H. pylori exhibiting antigenic properties have been shown to be useful in the detection of anti-H. pylori adhesive protein antibodies (Fauchere, J.L., and Blaser, M.J., Microbiol . Path ⁇ g . , 9:427-39 (1990)). Therefore, indirect ELISA detection of antibodies specific to H. pylori adhesive proteins is possible by means of incubating H. pyl ori extract with KATO III MEF coated on a microtitre plate. Adhesive proteins will adhere to KATO III MEF and will therefore be bound indirectly to the microtitre plate. The coated plate can subsequently be used to detect corresponding antibodies in human sera.

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Peptides Or Proteins (AREA)

Abstract

La présente invention concerne, non seulement une technique de diagnostic de l'infection d'un sujet par H. pylori, mais également des nécessaires à utiliser pour cette technique. L'invention concerne également des technique d'évaluation de l'état de l'affection gastrique du sujet, ainsi que des vaccins comprenant des antigènes spécifiques de protéines d'adhésines.
PCT/GB1997/002554 1996-09-20 1997-09-22 Adhesines obtenues d'heliobacter pylori et leurs utilisations therapeutiques et diagnostiques WO1998012562A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU43116/97A AU4311697A (en) 1996-09-20 1997-09-22 Adhesins from heliobacter pylori and their diagnostic and therapeutic uses

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
GB9619694.4 1996-09-20
GBGB9619694.4A GB9619694D0 (en) 1996-09-20 1996-09-20 Diagnostic method
GBGB9622846.5A GB9622846D0 (en) 1996-11-01 1996-11-01 Diagnostic method
GB9622846.5 1996-11-01

Publications (1)

Publication Number Publication Date
WO1998012562A1 true WO1998012562A1 (fr) 1998-03-26

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AU (1) AU4311697A (fr)
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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999049890A1 (fr) * 1998-03-31 1999-10-07 Daewoong Pharmaceutical Co., Ltd. VACCIN PREVENTIF ET THERAPEUTIQUE CONTRE LES MALADIES ASSOCIEES A $i(HELICOBACTER PYLORI)
US6617116B2 (en) 2000-01-28 2003-09-09 Genelabs Diagnostics Pte. Ltd. Assay devices and methods of analyte detection
US8475735B2 (en) 2004-11-01 2013-07-02 Uma Mahesh Babu Disposable immunodiagnostic test system

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993018150A1 (fr) * 1992-03-02 1993-09-16 Biocine S.P.A. Proteines d'helicobacter pylori utiles pour des vaccins et des diagnostics
WO1993022682A1 (fr) * 1992-04-29 1993-11-11 Auspharm International Limited Test in vitro pour helicobacter pylori
WO1996012965A1 (fr) * 1994-10-20 1996-05-02 Genelabs Diagnostics Pte Ltd. Methodes de diagnostic de l'helicobacter pylori et necessaires correspondants
DE19521314A1 (de) * 1995-06-12 1996-12-19 Max Planck Gesellschaft Adhärenzgen aus Helicobacter pylori und davon codiertes Polypeptid
DE19535321A1 (de) * 1995-09-22 1997-03-27 Max Planck Gesellschaft Neues Adhäsin aus Helicobacter pylori

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1993018150A1 (fr) * 1992-03-02 1993-09-16 Biocine S.P.A. Proteines d'helicobacter pylori utiles pour des vaccins et des diagnostics
WO1993022682A1 (fr) * 1992-04-29 1993-11-11 Auspharm International Limited Test in vitro pour helicobacter pylori
WO1996012965A1 (fr) * 1994-10-20 1996-05-02 Genelabs Diagnostics Pte Ltd. Methodes de diagnostic de l'helicobacter pylori et necessaires correspondants
DE19521314A1 (de) * 1995-06-12 1996-12-19 Max Planck Gesellschaft Adhärenzgen aus Helicobacter pylori und davon codiertes Polypeptid
DE19535321A1 (de) * 1995-09-22 1997-03-27 Max Planck Gesellschaft Neues Adhäsin aus Helicobacter pylori

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
A. MORAN: "Cell surface characteristics of Helicobacter pylori.", FEMS IMMUNOLOGY AND MEDICAL MICROBIOLOGY, vol. 10, no. 3-4, February 1995 (1995-02-01), AMSTERDAM, NL, pages 271 - 280, XP002052997 *
B. GERSTENECKER ET AL.: "Serodiagnosis of Helicobacter pylori infections with an enzyme immunoassay using the chromatographically purified 120 kilodalton protein.", EUROPEAN JOURNAL OF CLINICAL MICROBIOLOGY & INFECTIOUS DISEASES, vol. 11, no. 7, July 1992 (1992-07-01), BRAUNSCHWEIG, GERMANY, pages 595 - 601, XP000566365 *
C. SPIEGELHALDER ET AL.: "Purification of Helicobacter pylori superoxide dismutase and cloning and sequencing of the gene.", INFECTION AND IMMUNITY, vol. 61, no. 12, December 1993 (1993-12-01), WASHINGTON, DC, USA, pages 5315 - 5325, XP002052998 *
E. PESCI ET AL.: "Genetic organization and enzymatic activity of a superoxide dismutase from the microaerophilic human pathogen, Helicobacter pylori.", GENE, vol. 143, no. 1, 27 May 1994 (1994-05-27), AMSTERDAM, NL, pages 111 - 116, XP002052999 *
M. KHIN ET AL.: "Immunological detection of Helicobacter pylori in pregnant women.", BIOMEDICAL LETTERS, vol. 50, 1994, CAMBRIDGE, GB, pages 71 - 78, XP002052995 *
P. DOIG ET AL.: "Production of a conserved adhesin by the human gastroduodenal pathogen Helicobacter pylori.", JOURNAL OF BACTERIOLOGY, vol. 174, no. 8, April 1992 (1992-04-01), BALTIMORE, MD, USA, pages 2539 - 2547, XP002052996 *

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1999049890A1 (fr) * 1998-03-31 1999-10-07 Daewoong Pharmaceutical Co., Ltd. VACCIN PREVENTIF ET THERAPEUTIQUE CONTRE LES MALADIES ASSOCIEES A $i(HELICOBACTER PYLORI)
US6617116B2 (en) 2000-01-28 2003-09-09 Genelabs Diagnostics Pte. Ltd. Assay devices and methods of analyte detection
US6849414B2 (en) 2000-01-28 2005-02-01 Genelabs Diagnostics Pte Ltd. Assay devices and methods of analyte detection
US8475735B2 (en) 2004-11-01 2013-07-02 Uma Mahesh Babu Disposable immunodiagnostic test system

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Publication number Publication date
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