WO1998042372A1 - CARBOHYDRATE ANTIGENS WHICH IMMUNOREACT WITH POLYCLONAL ANTISERA AGAINST $i(H. PYLORI) - Google Patents
CARBOHYDRATE ANTIGENS WHICH IMMUNOREACT WITH POLYCLONAL ANTISERA AGAINST $i(H. PYLORI) Download PDFInfo
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- WO1998042372A1 WO1998042372A1 PCT/US1998/005850 US9805850W WO9842372A1 WO 1998042372 A1 WO1998042372 A1 WO 1998042372A1 US 9805850 W US9805850 W US 9805850W WO 9842372 A1 WO9842372 A1 WO 9842372A1
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- Prior art keywords
- antigen
- pylori
- polynucleotide
- carbohydrate
- clone
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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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/04—Antibacterial agents
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Definitions
- This invention relates to antigens and the use and production thereof. More particularly, the present invention is directed to carbohydrate antigens which immunoreact with polyclonal antisera against H. pylori .
- this invention relates to carbohydrate antigens which are derived from H. pylori .
- H. pylori Helicobacter pylori has been implicated as being a cause of peptic ulcer as a result of an infectious disease caused by H. pylori .
- antigens as well as polynucleotides encoding polypeptide ⁇ , which produce such antigens, which antigens immunoreact with H. pylori antisera.
- carbohydrate antigens as well as fragments and analogs thereof , each of which is immunoreactive with H. pylori carbohydrate specific polyclonal antisera.
- a procedure for producing such an antisera is described in the Example.
- the antigens of the present invention are produced in bacteria by polypeptides encoded by DNA present in clones which are part of ATCC Deposit No. 98373.
- bacteria produce carbohydrates, and such carbohydrates are produced in the bacteria by so-called gene clusters (a plurality of genes) which express polypeptides which interact in the bacteria to produce carbohydrates .
- the present invention in part, is directed to polynucleotides which form a gene cluster or operon, which, in bacteria, in particular E. coli . express polypeptides which in the bacteria produce carbohydrate antigens which immunoreact with H. pylori specific antisera.
- the gene cluster or operon is one which is recovered from H. pylori .
- Supercos 1 Stratagene
- antigens which immunoreact with H. pylori carbohydrate polyclonal antisera which antigens are produced by clones Ml, M2 and M3 contained in the deposited material.
- the present invention is directed to certain antigens produced by a clone deposited as ATCC Deposit No. 98373, which antigens immunoreact with H. pylori carbohydrate specific polyclonal antisera, as well as fragments and/or analogs of such antigens, as well as the polynucleotides forming a gene cluster which produce such antigens, analogs and fragments, which polynucleotides may be DNA and/or RNA.
- the DNA may be double-stranded or single-stranded, and if single stranded, may be the coding strand or non-coding (anti-sense) strand.
- the coding sequence of the gene cluster may be identical to that of a deposited clone or may be a different coding sequence which coding sequence, as a result of the redundancy or degeneracy of the genetic code, encodes the same polypeptides which generate the carbohydrate antigens which immunoreact with H. pylori antiserum as the DNA of the deposited DNA, or an immunoreactive analog or fragment of such antigen.
- the polynucleotide which is the gene cluster which generates an antigen which reacts with H. pylori antiserum may include : only the coding sequence , such coding sequence and additional coding sequence such as a leader or secretory sequence or a proprotein sequence; the coding sequence (and optionally additional coding sequence) and non-coding sequence, such as sequence 5' and/or 3' of the coding sequence or a fragment or portion of such polynucleotide which forms the gene cluster.
- polynucleotide encoding a polypeptide or “polynucleotide forming a gene cluster” encompasses a polynucleotide which includes only coding sequence as well as a polynucleotide which includes additional coding and/or non-coding sequence.
- the present invention further relates to variants of the hereinabove described polynucleotides which form a gene cluster which produces fragments, analogs and derivatives of the carbohydrate antigens which react with H. pylori antisera.
- the variant of the polynucleotide may be a naturally occurring allelic variant of the polynucleotide or a non-naturally occurring variant of the polynucleotide .
- the present invention also relates to polynucleotide probes constructed from a portion of the polynucleotide of the invention.
- Fragments of the polynucleotides of the present invention may be used as a hybridization probe for a library to isolate the full length DNA and to isolate other DNAs which have a high sequence similarity to the genes or portion of the genes forming the gene cluster.
- Probes of this type preferably have at least 10, preferably at least 15, and even more preferably at least 30 bases and may contain, for example, at least 50 or more bases.
- the probe may also be used to identify a DNA clone corresponding to a full length transcript and a genomic clone or clones that contain the complete gene including regulatory and promoter regions.
- An example of a screen comprises isolating the coding region of a gene by using a DNA sequence of a deposited clone, which is part of the operation to synthesize an oligonucleotide probe.
- Labeled oligonucleotides having a sequence complementary to that of a polynucleotide of the present invention are used to screen a library of genomic DNA to determine which members of the library the probe hybridizes to.
- the present invention further relates to polynucleotides which hybridize to the hereinabove- described sequences if there is at least 70%, preferably at least 90%, and more preferably at least 95% identity between the sequences in a complementary sense .
- the present invention particularly relates to polynucleotides which hybridize under stringent conditions to the hereinabove-described polynucleotides .
- stringent conditions means hybridization will occur only if there is at least 95% and preferably at least 97% identity in a complementary sense between the sequences.
- the polynucleotide may have at least 15 bases, preferably at least 30 bases, and more preferably at least 50 bases which hybridize to any part of a polynucleotide of the present invention and which has an identity thereto, as hereinabove described, and which may or may not retain activity.
- the present invention is directed to polynucleotides having at least a 70% identity, preferably at least 90% identity and more preferably at least a 95% identity to one or more polynucleotides which form the gene cluster for generating a carbohydrate antigen which immunoreacts with H. pylori antisera as well as fragments thereof, which fragments have at least 15 bases, preferably at least 30 bases and most preferably at least 50 bases, which fragments are at least 90% identical, preferably at least 95% identical and most preferably at least 97% identical to any portion of a polynucleotide of the present invention.
- identity when comparing a second sequence to a first sequence, means that the two sequences are properly aligned and then the second sequence is compared with the first sequence over the length of the first sequence. If any base of the first sequence does not match an aligned base in the second sequence or if the second sequence or the first sequence over the compared aligned length has a base which is not aligned with a base of the other sequence, then each of such occurrences is counted as a difference . The percent identity is then calculated by use of a fraction having as a numerator the difference between the number of bases in the first sequence and the number of base differences and as a denominator the number of bases in the first sequence.
- the present invention further relates to antigens which immunoreact with H. pylori antisera and which is produced by one of the deposited clones, as well as fragments, analogs and derivatives of such antigen.
- the antigens and polynucleotides of the present invention are preferably provided in an isolated form, and preferably are purified to homogeneity.
- isolated means that the material is removed from its original environment (e.g., the natural environment if it is naturally occurring) .
- a naturally-occurring polynucleotide or antigen present in a living animal is not isolated, but the same polynucleotide or antigen, separated from some or all of the coexisting materials in the natural system, is isolated.
- Such polynucleotides could be part of a vector and/or such polynucleotides or antigens could be part of a composition, and still be isolated in that such vector or composition is not part of its natural environment .
- the antigens of the present invention include the immunoreactive antigens produced by DNA of the deposited clones as well as fragments and derivatives thereof which immunoreac ive with H. pylori antigen.
- the present invention also relates to vectors which include polynucleotides of the present invention, host cells which are genetically engineered with polynucleotides or vectors of the invention and the production of antigens of the invention by recombinant techniques.
- Host cells are genetically engineered (transduced or transformed or transfected) with the vectors of this invention which may be, for example, a cloning vector or an expression vector.
- the vector may be, for example, in the form of a plasmid, a phage, etc.
- the engineered host cells can be cultured in conventional nutrient media modified as appropriate for activating promoters, selecting transfor ⁇ nants or amplifying the genes of the present invention.
- the culture conditions such as temperature, pH and the like, are those previously used with the host cell selected for expression, and will be apparent to the ordinarily skilled artisan.
- the polynucleotides of the present invention may be employed for producing antigens of the invention by recombinant techniques.
- the polynucleotide may be included in any one of a variety of expression vectors for expressing a polypeptide.
- Such vectors include chromosomal, nonchromosomal and synthetic DNA sequences, e.g., bacterial plasmids; phage DNA; etc.
- any other vector may be used as long as it is replicable and viable in the host.
- the appropriate DNA sequence may be inserted into the vector by a variety of procedures .
- the DNA sequence is inserted into an appropriate restriction endonuclease site(s) by procedures known in the art. Such procedures and others are deemed to be within the scope of those skilled in the art.
- the DNA sequence in the expression vector is operatively linked to an appropriate expression control sequence (s) (promoter) to direct mRNA synthesis.
- s expression control sequence
- promoters there may be mentioned: the E. coli. lac or trp, the phage lambda P L promoter and other promoters known to control expression of genes in prokaryotic cells.
- the expression vector also contains a ribosome binding site for translation initiation and a transcription terminator.
- the vector may also include appropriate sequences for amplifying expression.
- the expression vectors preferably contain one or more selectable marker genes to provide a phenotypic trait for selection of transformed host cells such as dihydrofolate reductase or neomycin resistance for eukaryotic cell culture, or such as tetracycline or ampi- cillin resistance in E. coli.
- the vector containing the appropriate DNA sequence as hereinabove described, as well as an appropriate promoter or control sequence, may be employed to transform an appropriate host to permit the host to express the protein.
- bacterial cells such as E. coli, Streptomvces , Bacillus subtilis .
- the selection of an appropriate host is deemed to be within the scope of those skilled in the art from the teachings herein.
- the present invention also includes recombinant constructs comprising one or more of the sequences as broadly described above.
- the constructs comprise a vector, such as a plasmid, into which a sequence of the invention has been inserted, in a forward or reverse orientation.
- the construct further comprises regulatory sequences, including, for example, a promoter, operably linked to the sequence .
- a promoter operably linked to the sequence .
- the following vectors are provided by way of example; pQE70, pQE60, pQE-9 (Qiagen) , pDIO, psiX174, pBluescript II KS, pNH8A, pNH16a, pNH18A, pNH46A (Stratagene) ; ptrc99a, pKK223-3, pKK233-3, pDR540, pRIT5 (Pharmacia) .
- any other plasmid or vector may be used as long as they are replicable and viable in the host.
- the present invention relates to host cells containing the above-described constructs, in particular a bacterial cell.
- Introduction of the construct into the host cell can be effected by calcium phosphate transfection, DEAE-Dextran mediated transfection, or electroporation (Davis, L., Dibner, M., Battey, I., Basic Methods in Molecular Biology, (1986) ) .
- the antigens of the present invention which immunoreact with H. pylori carbohydrate polyclonal antisera, and which are produced by a deposited clone, as well as their fragments, derivatives or analogs, may be used as an immunogen to produce antibodies against H. pylori .
- These antibodies can be, for example, polyclonal or monoclonal antibodies .
- the present invention also includes chimeric, humanized, and single-chain antibodies, as well as fragments, thereof. Such antibodies may be produced by procedures generally known in the art .
- the antigens or antibodies generated against the antigens may be employed in immunoassays of the type known in the art for detecting the presence of H. pylori and/or antibodies against H. pylori .
- the antigens may be employed to induce in a host an immune reaction against H. pylori , preferably in the form of an immunogen wherein the antigen is combined with an appropriate protein.
- the antigen is conjugated to a protein which is known to induce a CD4 T-cell response.
- a protein which is known to induce a CD4 T-cell response.
- proteins are known in the art and have been previously employed in the art to induce a T cell response and thereby T-cell dependent memory.
- Preferred proteins are diphtheria or tetanus toxoids but other proteins having such properties are within the scope of the invention and should be apparent to those skilled in the art from the teachings herein.
- the carbohydrate antigen may be directly conjugated to the protein or through an appropriate linker or spacer.
- the antigens of the present invention may be employed to induce an immune response, in a host, against H. pylori .
- the host is a mammal, which may be human or non-human.
- Such antigens and polynucleotides are preferably employed to induce a protective immune response against H. pylori infection and, therefore, may be employed for treatment and/or prevention of H. pylori infection.
- An antigen of the present invention when used for such a purpose, is generally combined with a pharmaceutically effective carrier and is administered in an amount effective to reduce and/or prevent H. pylori infection.
- a pharmaceutically effective carrier In general, such polypeptide would be administered in an amount from 10 ⁇ g to 500 mg, and such administration, for example, may be parenteral, oral or intranasal administration.
- a polynucleotide, forming a gene cluster of the present invention is employed to produce a recombinant, non-toxigenic organism; in particular, a bacterial organism, which produces an antigen of the present invention.
- a recombinant, non-toxigenic organism in particular, a bacterial organism, which produces an antigen of the present invention.
- V. cholerae and S . typhi As representative examples of such organisms, there may be mentioned V. cholerae and S . typhi .
- a wild-type strain of V. cholerae or S. typhi may be attenuated by procedures known in the art to produce a live bacterial strain which has been attenuated to render the bacteria non-toxigenic.
- Such non-toxigenic recombinant strains may be employed as a live vaccine for producing, in vivo, an antigen of the present invention to induce a protective immune response.
- the non-toxigenic V. cholerae which is employed for producing a recombinant non-toxigenic V. cholerae in accordance with the present invention is V. cholerae of the type which has been developed for potential use as a vaccine against V. cholerae.
- V. cholerae is a live, attenuated V. cholerae which has been attenuated to render such V. cholerae non-toxigenic.
- the V. cholerae which is employed for producing a recombinant vaccine in accordance with the present invention may be an attenuated 01 strain or a non-01 strain.
- wild-type V. cholerae strains are attenuated for example, to render same non-toxigenic by a genetic deletion which removes three toxins (cholerae toxin, zonula occludens toxin, and accessory cholerae enterotoxin) , and which may be further attenuated by removal of a colonization factor (core encoded pilin) , and RSI (a site-specific toxin acquisition cassette) .
- a colonization factor core encoded pilin
- RSI site-specific toxin acquisition cassette
- the strains hereinabove described were engineered to reintroduce into the recA locus the gene encodings ctxB, which yields the strains referred to as Peru-3 and Bengal-3.
- Non-toxigenic strain of V. cholerae which is a spontaneous non-motile filamentous isolate of Peru-3 has also been isolated as designated as Peru-14.
- Non-motile isolates of Peru-3 and Bengal-3 have also been identified which have been designated as Peru-15 and Bengal-15.
- Non-toxigenic strains of V. cholerae are generally known in the art, and such strains may be employed in the present invention.
- Such strains include but are not limited to: Bengal-2, Bengal -3 , Bengal -15, Peru-2, Peru- 3, Peru-14 , Peru-15, Bah-2, Bah-15, Bang-2, Bang-3 , and Bang- 15.
- a non-toxigenic strain of V. cholerae is transformed with a gene cluster of the invention which produces in the organism a carbohydrate antigen which induces an immune response against H. pylori.
- the polynucleotide is incorporated into a suitable vector for transformation of the V. cholerae.
- Such vector may be one which integrates the polynucleotide into the chromosome of V. cholerae or may be one which provides for extrachromosomal transformation.
- the present invention contemplates a recombinant non-toxigenic V. cholerae in which a polynucleotide gene cluster produces a carbohydrate antigen which induces an immune response against H. pylori is present either extrachromosomally or integrated into the chromosome of the V. cholorae.
- a polynucleotide gene cluster of the invention is placed in an appropriate vector, and such vector is employed for transforming a non-toxigenic strain of V. cholerae to provide a vaccine for use in the treatment and/or prevention of H. pylori infection.
- the vector is provided with an appropriate expression promoter.
- the vector which is employed is one which functions in V. cholerae to provide for expression of the polypeptides which in the organism produce the carbohydrate antigen which induces an immune response against H. pylori .
- the promoter in a preferred embodiment, is derived from V. cholerae.
- a preferred promoter is a V. cholerae heat shock promoter known as htpGp .
- the vector or plasmid generally also includes a suitable marker to permit selection of recombinant strains .
- Non-toxigenic V. cholerae which has been transformed to produce a carbohydrate antigen which provides an immune response against H. pylori and in particular a protective immune response may be employed as an oral vaccine for treatment and/or prevention of H. pylori infection.
- the vaccine is employed in an amount effective to treat or prevent H. pylori infection.
- the recombinant V. cholerae is administered in a dosage of from about 10 7 to 10 8 colony forming units.
- the vaccine may be administered in a single dose or in multiple doses; e.g., biweekly.
- the recombinant V. cholerae in accordance with the present invention may be incorporated into an appropriate pharmaceutically acceptable carrier and in particular for oral delivery.
- the carrier may be a liquid; e.g., and mixture of bicarbonate and ascorbate or a solid such as an enteric coated capsule.
- the non-toxigenic V. cholerae is transformed by use of an appropriate vector, which in one embodiment integrates the polynucleotide expressing the appropriate antigen or immunogen into the chromosome of the V. cholerae .
- an appropriate vector which in one embodiment integrates the polynucleotide expressing the appropriate antigen or immunogen into the chromosome of the V. cholerae .
- Such vector or plasmid may be employed to facilitate site specific integration onto the chromosome; for example, at either the lacZ, recA or irgA loci (Published PCT Application WO 94/01780) .
- the vector when integration is desired, is capable of inserting the polynucleotide into the V. cholerae chromosome at the lacZ locus .
- Integrating vectors may also be employed for providing transformation in which the appropriate polypeptides are expressed from extrachromosomal DNA.
- an appropriate selection procedure is employed to select V. cholerae in which the polynucleotide has been integrated into the chromosome or V. cholerae in which the DNA is maintained as an extrachromosomal plasmid.
- an attenuated V. cholerae in which the recA gene is deleted is used for preparation of a recombinant V. cholerae in which the polypeptides are expressed from an extrachromosomal plasmid.
- an attenuated V. cholerae which includes the recA gene is used for preparation of a recombinant V. cholerae in which the polypeptides are expressed from DNA integrated into the V. cholerae chromosome .
- the same vector may be employed, e.g., a plasmid which includes a lacZ sequence.
- the recA gene is deleted, the polynucleotide is not integrated into the chromosome; however, when such vector is used for transformation of attenuated V. cholerae which includes recA gene, integration can occur at the lacZ locus and a recombinant V. cholerae in which the DNA is integrated at the lacZ locus can be selected.
- a product and a process for treatment and/or prevention of H. pylori infection wherein a non-toxigenic V. cholerae strain is transformed with a polynucleotide which encodes a polypeptide of the invention which provides an immune response against H. pylori .
- a non-toxigenic V. cholerae strain is transformed with a polynucleotide which encodes a polypeptide of the invention which provides an immune response against H. pylori .
- Such recombinant non-toxigenic V. cholerae strain may be employed as a live oral vaccine for the treatment and/or prevention of H. pylori infection.
- more than one copy of a polynucleotide encoding a polypeptide which induces an immune response may be integrated into the chromosome of V. cholerae .
- such polynucleotide may be integrated into more than one locus in the chromosome; e.g., lacZ, recA, and irgA.
- Such recombinant products may also be employed in vi tro to produce the carbohydrate antigen.
- H. pylori Claudio strain was streaked on Trypticase Soy agar supplemented with 5% defibrinated sheep blood and incubated at 37°C for 48 to 72h in an anaerobic chamber. Cells were collected from plates with a sterile swab and suspended in l ml Brucella broth. The broth culture was deemed pure by: i) colonial morphology, ii) wet mount microscopy, iii) Gram stain, iv) urease + , and, v) agglutination of cells with anti-whole-cell helicobacter polyclonal rabbit sera. The characterized Claudio strain was cell-banked in 10% glycerol and stored in Cryovials in liquid nitrogen.
- Chromosomal DNA was digested with Sau3A and BamHI and electrophoresed on 0.7% agarose gel .
- Southern blot analysis of chromosomal DNA probed with a H. pylori ureAB gene fragment exhibited a strong hybridization signal indicating that the chromosomal DNA was H. pylori in origin.
- Carbohydrate was prepared by hot phenol- water extraction and purified by ultra-centrifugation.
- SDS- PAGE/silver staining analysis revealed 0-antigen banding patterns very similar to those observed by Mills et al (1) .
- Polysaccharide was quantitated by dry weight in order to determine polysaccharide amounts in subsequent immunization and ELISA experiments .
- a DNA cosmid library of H. pylori Cl chromosomal DNA was made using the SuperCosl cosmid vector (Stratagene) . Chromosomal DNA was partially digested with Sau3A for 3 min. and phosphatased. Vector SuperCosl cosmid DNA was prepared by digestion with Xbal, phosphatase treatment, and digestion with BamHI . Insert and vector DNA were ligated at 4°C overnight and packaged into Gigapacklll Gold packaging extract per manufacturers instructions. Titration of the unamplified cosmid library in E. coli XL1B- MR was 3.4xl0 4 cfu/ml. Amplification of the cosmid library generated clones at 5xl0 7 cfu/ml. The efficiency of packaging was calculated and in accord with the value indicated in the manufacturer's manual.
- a rabbit was immunized intramuscularly with the characterized, purified Claudio polysaccharide.
- the priming dose was 50ug polysaccharide followed by 3 boosts of lOOug polysaccharide over a period of 2 weeks.
- the sera was adsorbed twice with E. coli XLIB-MR strain (host strain used to generate the Claudio DNA cosmid library) .
- Silver staining resul ts Polysaccharide was prepared from cosmid recombinants by the hot water-phenol extraction procedure (3) . Briefly, cells grown either in broth culture or on agar plates were treated with hot water-phenol, dialyzed, and lyophilized . Results of silver staining of polysaccharide harvested from washed cells grown in LB broth or unwashed cells grown on agar plates indicated no reproducible difference between E. coli and the cosmid recombinants .
- EPS extra-cellular polysaccharide
- Polysaccharides of several Gram-negative bacteria have been shown to adhere to mammalian cells, including erythrocytes (RBC) . This property is considered a virulence feature of pathogens .
- Bacteria in which adherence to cells has been shown to be mediated by external polysaccharides include Shigella dysenteriae type 1 (4) , Campylobacter jejuni (5) , Vibrio cholerae (6) , Escherichia coli (7) .
- HA hemagglutination
- the carbohydrate concentration (14) of purified EPS derived from recombinant clones was, M, (5.6mg/ml), M 2 (7.6mg/ml), and M 3 (5.6mg/ml), >2-fold more carbohydrate than E. coli XL1B (2.8mg/ml).
- EPS was dissolved in distilled water and tested for HA titer.
- EPS of M lf M 2 , M 3 and E. coli XL1B exhibited HA titers of 4 , 32, 64 and 0 respectively, similar values to the E. coli cosmid supernatants described above.
- HAI Inhibition of HA
- sRBC constant amounts of sRBC and antigen (4-fold excess antigen of the calculated HA titer) and a titration of sera.
- the titer was 1:32, therefore a 1:8 dilution of supernatant was added to sRBC.
- HAI was observed when supernatants of Claudio and M 2 and M 3 were pre-treated with titrated amounts of anti-polysaccharide rabbit serum.
- a whole-cell ELISA was performed to further characterize M 2 and M 3 .
- the ELISA was done using whole-cells of H. pylori Claudio, M 2 and M 3 , and E. coli .
- Cells were grown on LB agar, resuspended in PBS, and bound to microtiter wells (180 ul/well) at concentrations ranging from 10 4 cfu/ml to 10 9 cfu/ml .
- Anti-polysaccharide rabbit sera was used as the primary antibody at 1:15 dilution and the conjugate was anti-rabbit IgG-alkaline phosphatase.
- M 2 and M 3 were highly immunopositive at cell numbers ranging from 1.8xl0 6 -1.8xl0 8 .
- the E. coli cells bound to the well, negative control was of negligible OD value at all cell numbers tested.
- the anti-polysaccharide sera used to screen the cosmid library and identify the clones is immunoreactive with M 2 and M 3 by ELISA.
- a polysaccharide ELISA was performed to determine the titer of anti-polysaccharide rabbit sera against Claudio polysaccharide.
- An Immulon 1 microtiter plate was coated with 2ug/well Claudio polysaccharide .
- Titrated anti-Claudio polysaccharide rabbit serum was used as the primary antibody and a monoclonal anti-rabbit Ig-alkaline phosphatase as the secondary antibody/conjugate.
- the rabbit sera exhibited an ELISA titer of >1:5,000.
- Polysaccharide extracted from M 2 and M 3 clones grown on LB agar plates used in an identical ELISA displayed baseline titers, similar to that of E. coli .
- a competitive ELISA was employed to determine whether supernatants or whole cell antigens derived from Claudio and cosmid recombinants M 2 and M 3 could inhibit the interaction between anti-Claudio polysaccharide antibodies and polysaccharide antigen (section 5B above) .
- An Immulon 1 microtiter plate was coated with 2ug/well Claudio polysaccharide.
- Anti-Claudio polysaccharide sera (1:50) was pre-incubated with two-fold, serial dilutions of antigen containing supernatant at 37°C for lh.
- Motility test of the Ml-3 clones and E. coli were also observed on solid agar by mixing cells with 30 ml Luria- Bertani soft agar (0.3% agar) and poured into a petri dish. After 1 h, 15 ml LB agar (1.5%) was poured on top of the solidified soft agar and plates were incubated overnight at 37°C followed by room temperature incubation for 5 days.
- Ml-3 exhibited significantly slower motility in soft agar compared to the E. coli .
- M-clones were suspended in PBS and observed microscopically, significantly slower motility was observed compared to the cosmid bearing host E. coli .
- EPS Exopolysaccharide
- Exopolysaccharide (EPS) samples of clones Ml-3, H. pylori Cl and E. coli were tested for both neutral and acidic sugars .
- EPS samples were hydrolyzed in 2M TFA at 121°C for 2 h.
- the hydrolyzed samples were reduced by sodium borodeuteride and acetylated using acetic anhydride and pyridine. Myo-ionositol was added as an internal standard.
- the samples were run on a GC-MS using a Sp2330 Supelco column.
- acidic sugar analysis the samples were hydrolyzed with freshly prepared 1M methanolic-HCL for 16 h at 80°C.
- the released sugars were derivatized with Tri-Sil and the samples were run on GC using the Supelco column.
- the rEPS clones had significantly higher amounts of fucose in comparison to E. coli, while mannose concentrations of the clones were 4-5 fold less in comaprison to E. coli.
- the mole percent amounts of fucose and mannose from the rEPS clones resembles that of H. pylori , differing considerably from E. coli and the fucose/mannose ratio of the rEPS clones resembles that of strain Cl rather than E. coli .
- the other sugars were present in similar amounts (Table-I) .
- the specific region of the H. pylori DNA contained in the deposited clones which form the gene cluster which produces the carbohydrate antigen is determined by sub-cloning of the cosmid DNA derived from recombinant clones M,, M 2 , and M 3 .
- Cosmid DNA is purified by Qiagen midi-prep columns, and subsequently digested with restriction endonucleases (e.g., EcoRl, Bgll, Hindlll, etc.).
- DNA fragments resulting from such digestions are ligated into a cloning vector (e.g., pBluescript II KS, pKK223-3, pUC19, etc.) and digested with the corresponding restriction endonuclease .
- the products of the ligations are transformed into E. coli XLIB-MR.
- Recombinant clones are tested for production of the carbohydrate antigen by colony immunoblot as described previously.
- the immunoreactive clones are isolated, purified, and screened for phenotypic expressions such as mucoidy on solid agar and agglutination of sRBC by their supernatant or whole cells. This sub-cloning process determines the minimum size of the H. pylori DNA included in a deposited clone for producing in bacteria the carbohydrate antigen which reacts with H. pylori anti sera. The subcloned DNA is then sequenced.
- the deposit made to American Type Culture Collection is a mixture of 3 recombinant clones of Escherichia coli XLIB-MR (:M,, M 2 , and M 3 ) which harbor cosmids expressing Helicobacter pylori carbohydrate antigen.
- Cells were grown in Luria Bertani (LB) broth supplemented with ampicillin (50 ug/ml) with glycerol added to final concentration of 15%.
- LB Luria Bertani
- ampicillin 50 ug/ml
- glycerol added to final concentration of 15%.
- a sample of the frozen vial should be streak plated for individual colonies on LB agar supplemented with ampicillin (50 ug/ml) and incubated at 37°C for 18 hr. On agar plates, colonies are mucoid in appearance.
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Priority Applications (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP98912029A EP0971734A4 (en) | 1997-03-26 | 1998-03-25 | Carbohydrate antigens which immunoreact with polyclonal antisera against h. pylori |
CA002284671A CA2284671A1 (en) | 1997-03-26 | 1998-03-25 | Carbohydrate antigens which immunoreact with polyclonal antisera against h. pylori |
AU65842/98A AU6584298A (en) | 1997-03-26 | 1998-03-25 | Carbohydrate antigens which immunoreact with polyclonal antisera against (h. pylori) |
JP54594998A JP2001520671A (en) | 1997-03-26 | 1998-03-25 | Carbohydrate antigen immunoreacts with polyclonal antiserum against Helicobacter pylori |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
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US4240397P | 1997-03-26 | 1997-03-26 | |
US4397697P | 1997-04-23 | 1997-04-23 | |
US60/042,403 | 1997-04-23 | ||
US60/043,976 | 1997-04-23 |
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WO1998042372A1 true WO1998042372A1 (en) | 1998-10-01 |
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PCT/US1998/005850 WO1998042372A1 (en) | 1997-03-26 | 1998-03-25 | CARBOHYDRATE ANTIGENS WHICH IMMUNOREACT WITH POLYCLONAL ANTISERA AGAINST $i(H. PYLORI) |
Country Status (5)
Country | Link |
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EP (1) | EP0971734A4 (en) |
JP (1) | JP2001520671A (en) |
AU (1) | AU6584298A (en) |
CA (1) | CA2284671A1 (en) |
WO (1) | WO1998042372A1 (en) |
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0899343A2 (en) * | 1997-07-24 | 1999-03-03 | University Of Bristol | Bacterial polysaccharide |
JP2004500806A (en) * | 1999-10-26 | 2004-01-15 | バイオロジカル・ターゲッツ・インコーポレーテッド | Microorganisms, microbial exopolysaccharides and their uses |
Families Citing this family (1)
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CN111467368B (en) * | 2020-04-13 | 2021-05-28 | 江南大学 | Application of oligosaccharide compound containing heptasaccharide chain in preparation of helicobacter pylori vaccine |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5514660A (en) * | 1992-07-31 | 1996-05-07 | Neose Pharmaceuticals, Inc. | Method for treating and inhibiting gastric and duodenal ulcers |
US5625124A (en) * | 1994-07-11 | 1997-04-29 | Washington University | Animal model for helicobacter pylori infection |
Family Cites Families (1)
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AR003125A1 (en) * | 1995-06-01 | 1998-07-08 | Astra Ab | BACTERIAL ANTIGENS FOR THE DIAGNOSIS OF INFECTIONS WITH HELICOBACTER PYLORI, A DNA MOLECLE THAT CODES IT, A VECTOR, A HOST CELL, A PROCEDURE FOR PRODUCING THE POLIPEPTIDE, USE OF ELEPIPETICO, AND PROAPILY USE |
-
1998
- 1998-03-25 WO PCT/US1998/005850 patent/WO1998042372A1/en not_active Application Discontinuation
- 1998-03-25 JP JP54594998A patent/JP2001520671A/en active Pending
- 1998-03-25 AU AU65842/98A patent/AU6584298A/en not_active Abandoned
- 1998-03-25 EP EP98912029A patent/EP0971734A4/en not_active Withdrawn
- 1998-03-25 CA CA002284671A patent/CA2284671A1/en not_active Abandoned
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US5514660A (en) * | 1992-07-31 | 1996-05-07 | Neose Pharmaceuticals, Inc. | Method for treating and inhibiting gastric and duodenal ulcers |
US5625124A (en) * | 1994-07-11 | 1997-04-29 | Washington University | Animal model for helicobacter pylori infection |
Non-Patent Citations (10)
Title |
---|
ALKOUT A. H., ET AL.: "IDENTIFICATION OF SURFACE COMPONENTS OF HELICOBACTER PYLORI THAT BIND H TYPE 2 AND LEWIS BLOOD GROUP ANTIGENS.", GUT, BRITISH MEDICAL ASSOCIATION, LONDON,, UK, vol. 37., no. SUPPL. 01., 1 January 1995 (1995-01-01), UK, pages A21., XP002912714, ISSN: 0017-5749 * |
AMERICAN SOCIETY FOR MICROBIOLOGY, WASHINGTON.; 4 May 1997 (1997-05-04), GOLDBERG J. B., ET AL.: "SEQUENCE ANALYSIS AND CHARACTERIZATION OF THE RFAC GENE FROM HELICOBACTER PYLORI.", XP002912713 * |
DICK W E. BEURRET M: "GLYCOCONJUGATES OF BACTERIAL CARBOHYDRATE ANTIGENS A SURVEY AND CONSIDERATION OF DESIGN AND PREPARATION FACTORS", CONTRIBUTIONS TO MICROBIOLOGY AND IMMUNOLOGY., S. KARGER AG, BASEL, CH, vol. 10, 1 January 1989 (1989-01-01), CH, pages 48 - 114, XP002912720, ISSN: 0301-3081 * |
DROUET E. B., ET AL.: "PARTIAL CHARACTERIZATION OF AN EXTERNAL POLYSACCHARIDE OF HELICOBACTER PYLORI BY USING AN IMMUNOGLOBULIN M MONOCLONAL ANTIBODY.", INFECTION AND IMMUNITY, AMERICAN SOCIETY FOR MICROBIOLOGY., US, vol. 61., no. 06., 1 June 1993 (1993-06-01), US, pages 2732 - 2736., XP002912715, ISSN: 0019-9567 * |
GOLDBERG J B, ET AL: "CLONING OF THE LIPOPOLYSACCHARIDE (LPS)-CORE GENE RFAC, FROM HELICOBACTER PYLORI", STN BIOSIS, XX, XX, 1 January 1900 (1900-01-01), XX, XP002912712 * |
MILLS S. D., ET AL.: "ANTIGENICITY OF HELICOBACTER PYLORI LIPOPOLYSACCHARIDES.", JOURNAL OF CLINICAL MICROBIOLOGY, AMERICAN SOCIETY FOR MICROBIOLOGY, US, vol. 30., no. 12., 1 December 1992 (1992-12-01), US, pages 3175 - 3180., XP002912716, ISSN: 0095-1137 * |
MORAN A. P., ET AL.: "COMPOSITIONAL ANALYSIS OF HELICOBACTER PYLORI ROUGH-FORM LIPOPOLYSACCARIDES.", JOURNAL OF BACTERIOLOGY, AMERICAN SOCIETY FOR MICROBIOLOGY, US, vol. 174., no. 04., 1 February 1992 (1992-02-01), US, pages 1370 - 1377., XP002912717, ISSN: 0021-9193 * |
PIOTROWSKI J., ET AL.: "INHIBITION OF HELICOBACTER PYLORI COLONIZATION BY SULFATED GASTRIC MUCIN.", BIOCHEMISTRY INTERNATIONAL., ACADEMIC PRESS, LONDON., GB, vol. 24., no. 04., 1 July 1991 (1991-07-01), GB, pages 749 - 756., XP002912718, ISSN: 0158-5231 * |
RAPPUOLI R, ET AL.: "DEVELOPMENT OF A VACCINE AGAINST HELICOBACTER PYLORI: A SHORT OVERVIEW", ITALIAN JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY, PACINI EDITORE, PISA, IT, vol. 05, no. 02, 1 January 1993 (1993-01-01), IT, pages 576 - 578, XP002912719, ISSN: 1125-8055 * |
See also references of EP0971734A4 * |
Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0899343A2 (en) * | 1997-07-24 | 1999-03-03 | University Of Bristol | Bacterial polysaccharide |
EP0899343A3 (en) * | 1997-07-24 | 1999-08-11 | University Of Bristol | Bacterial polysaccharide |
JP2004500806A (en) * | 1999-10-26 | 2004-01-15 | バイオロジカル・ターゲッツ・インコーポレーテッド | Microorganisms, microbial exopolysaccharides and their uses |
Also Published As
Publication number | Publication date |
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EP0971734A1 (en) | 2000-01-19 |
EP0971734A4 (en) | 2001-02-07 |
CA2284671A1 (en) | 1998-10-01 |
JP2001520671A (en) | 2001-10-30 |
AU6584298A (en) | 1998-10-20 |
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